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	<title>bioRxiv Channel: Francis Crick Institute</title>
	<link>https://biorxiv.org</link>
	<description>
	This feed contains articles for bioRxiv Channel "Francis Crick Institute"
	</description>

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	<prism:publicationName>bioRxiv</prism:publicationName>
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	<title>bioRxiv</title>
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	<link>https://biorxiv.org</link>
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	<item rdf:about="https://biorxiv.org/cgi/content/short/078204v1?rss=1">
<title>
<![CDATA[
Myosin II controls junction fluctuations to guide epithelial tissue ordering 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/078204v1?rss=1"
</link>
<description><![CDATA[
Homophilic interactions between E-Cadherin molecules generate adhesive interfaces or junctions (AJs) that connect neighbouring cells in epithelial monolayers. These are highly dynamic structures. Under conditions of homeostasis, changes in the length of individual interfaces provide epithelia with the fluidity required to maintain tissue integrity in the face of cell division, delamination and extrinsic forces. Furthermore, when acted upon by polarized actomyosin-based forces, changes in AJ length can also drive neighbour exchange to reshape an entire tissue. Whilst the contribution of AJ remodelling to developmental morphogenesis has been subjected to intensive study, less is known about AJ dynamics in other circumstances. Here, using a combination of experiment and computational modelling, we study AJ dynamics in an epithelium that undergoes a gradual increase in packing order without concomitant large-scale changes in tissue shape or size. Under these conditions, we find that neighbour exchange events are driven by stochastic fluctuations in junction length, which are regulated at least in part by the level of junctional actomyosin. As a result of this behaviour, the steady increase in junctional actomyosin and consequent tension that accompanies development steadily reduces the rate of neighbour exchange and orders the tissue. This leads us to propose a model in which topological transitions, that underpin tissue fluidity, are either inhibited or biased by actomyosin-based forces, to drive, respectively, tissue ordering or deformation.
]]></description>
<dc:creator>Scott Curran</dc:creator>
<dc:creator>Charlotte Strandkvist</dc:creator>
<dc:creator>Jasper Bathmann</dc:creator>
<dc:creator>Marc de Gennes</dc:creator>
<dc:creator>Alexandre Kabla</dc:creator>
<dc:creator>Guillaume Salbreux</dc:creator>
<dc:creator>Buzz Baum</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-09-29</dc:date>
<dc:identifier>doi:10.1101/078204</dc:identifier>
<dc:title><![CDATA[Myosin II controls junction fluctuations to guide epithelial tissue ordering]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/075135v1?rss=1">
<title>
<![CDATA[
The FACT complex and cell cycle progression are essential to maintain asymmetric transcription factor partitioning during cell division 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/075135v1?rss=1"
</link>
<description><![CDATA[
The polarized partitioning of proteins in cells underlies asymmetric cell division, which is an important driver of development and cellular diversity. Like most cells, the budding yeast Saccharomyces cerevisiae divides asymmetrically to give two distinct daughter cells. This asymmetry mimics that seen in metazoans and the key regulatory proteins are conserved from yeast to human. A well-known example of an asymmetric protein is the transcription factor Ace2, which localizes specifically to the daughter nucleus, where it drives a daughter-specific transcriptional network. We performed a reverse genetic screen to look for regulators of asymmetry based on the Ace2 localization phenotype. We screened a collection of essential genes in order to analyze the effect of core cellular processes in asymmetric cell division. This identified a large number of mutations that are known to affect progression through the cell cycle, suggesting that cell cycle delay is sufficient to disrupt Ace2 asymmetry. To test this model we blocked cells from progressing through mitosis and found that prolonged cell cycle arrest is sufficient to disrupt Ace2 asymmetry after release. We also demonstrate that members of the evolutionary conserved FACT chromatin-remodeling complex are required for both asymmetric and cell cycle-regulated localization of Ace2.
]]></description>
<dc:creator>Eva Herrero</dc:creator>
<dc:creator>Sonia Stinus</dc:creator>
<dc:creator>Eleanor Bellows</dc:creator>
<dc:creator>Peter H Thorpe</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-09-19</dc:date>
<dc:identifier>doi:10.1101/075135</dc:identifier>
<dc:title><![CDATA[The FACT complex and cell cycle progression are essential to maintain asymmetric transcription factor partitioning during cell division]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/078196v1?rss=1">
<title>
<![CDATA[
Protein Kinase C and the stress response pathways are required for kinetochore homeostasis. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/078196v1?rss=1"
</link>
<description><![CDATA[
Kinetochores serve both a structural role linking chromosomes to the mitotic spindle and a regulatory role, controlling the timing of mitosis via the spindle assembly checkpoint. To identify proteins that regulate the kinetochore we used a genome-wide fluorescence microscopy approach. We combined an array of mutants of either non-essential gene deletions or essential temperature-sensitive alleles with fluorescently tagged spindle pole bodies (centrosome) and outer kinetochores. Quantitative and qualitative analysis revealed mutants that affect the levels and distribution of kinetochores respectively. These mutants are enriched for those involved in mRNA processing, chromatin organization, DNA replication/repair and mitosis. Our data show that the Pkc1 kinase maintains the kinetochore focus via its ability to prevent cell stress and this phenotype is rescued by an osmotic stabilizer. These data support the notion that kinetochore and microtubule homeostasis are perturbed by the stress response pathways. Hence this observation provides a candidate mechanism for extracellular stress leading to chromosome segregation defects.
]]></description>
<dc:creator>Elena Ledesma-Fernández</dc:creator>
<dc:creator>Eva Herrero</dc:creator>
<dc:creator>Guđjón Ólafsson</dc:creator>
<dc:creator>Peter Thorpe</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-09-28</dc:date>
<dc:identifier>doi:10.1101/078196</dc:identifier>
<dc:title><![CDATA[Protein Kinase C and the stress response pathways are required for kinetochore homeostasis.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/111658v1?rss=1">
<title>
<![CDATA[
Free ISG15 as a dimer generates IL-1β-producing CD8α+ dendritic cells at the site of infection 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/111658v1?rss=1"
</link>
<description><![CDATA[
ISG15 is strongly induced after type I IFN stimulation producing a protein comprised of two ubiquitin-like domains. Intracellularly, ISG15 can be covalently linked and modify the function of target proteins (ISGylation). In addition, free unconjugated ISG15 can be released from cells. We found that ISG15 is released in the serum of Toxoplasma gondii infected mice early after infection in a type-I IFN independent manner. Once in the extracellular space, free ISG15 forms dimers and enhances the release of key cytokines involved in the immune response to the parasite: IL-12, IFN-{gamma}, and IL-1{beta}. Its action is dependent on an actively invading and replicating live parasite. ISG15 induces an increase of IL-1{beta} later during infection by leading to increased IL-1{beta} producing CD8+ dendritic cells at the site of infection. Here, we define for the first time the molecular determinants of active free ISG15 and link ISG15 to IL-1{beta} production by CD8+ dendritic cells. Thus we define ISG15 as a novel secreted modulator of the cytokine response during Toxoplasma infection.
]]></description>
<dc:creator>Napolitano, A.</dc:creator>
<dc:creator>van der Veen, A. G.</dc:creator>
<dc:creator>Bunyan, M.</dc:creator>
<dc:creator>Borg, A.</dc:creator>
<dc:creator>Kjaer, S.</dc:creator>
<dc:creator>Beling, A.</dc:creator>
<dc:creator>Knobeloch, K.-P.</dc:creator>
<dc:creator>Frickel, E.</dc:creator>
<dc:date>2017-02-27</dc:date>
<dc:identifier>doi:10.1101/111658</dc:identifier>
<dc:title><![CDATA[Free ISG15 as a dimer generates IL-1β-producing CD8α+ dendritic cells at the site of infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-02-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/041988v1?rss=1">
<title>
<![CDATA[
A simple analytical formula to compute the residual Mutual Information between pairs of data vectors 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/041988v1?rss=1"
</link>
<description><![CDATA[
SummaryThe Mutual Information of pairs of data vectors, for example sequence alignment positions or gene expression profiles, is a quantitative measure of the interdependence between the data. However, data vectors based on a finite number of samples retain non-zero Mutual Information values even for completely random data, which is referred to as background or residual Mutual Information. Estimates of the residual Mutual Information have so far been obtained through heuristic or numerical approximations. Here we introduce a simple analytical formula for the computation of the residual Mutual Information that yields precise values and does not require the joint probabilities between the vector elements as input.nnAvailability and ImplementationA C program arMI is available at http://mathbio.crick.ac.uk/wiki/Software#arMI. Using an input alignment in FASTA format or alternatively an internally created random alignment of specified length and depth, the program computes three types of Mutual information: (i) Shannons Mutual Information between all pairs of alignment columns; (ii) the numerical residual Mutual Information by using the same formula on the randomised (shuffled) data; (iii) the analytical residual Mutual Information introduced here. The package depends on the GNU Scientific Library, which is used for vector and matrix operations, factorial expressions and random number generation (Galassi et al., 2009). Reference alignments and result data are included in the program package in the folder  tests. The R environment was used for statistics and plotting (R Core Team, 2014).nnContactJens.Kleinjung@crick.ac.uknnSupplementary MaterialA detailed derivation of the analytical formula is given in the Supplementary Material.
]]></description>
<dc:creator>Jens Kleinjung</dc:creator>
<dc:creator>Anthony C.C. Coolen</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-03-01</dc:date>
<dc:identifier>doi:10.1101/041988</dc:identifier>
<dc:title><![CDATA[A simple analytical formula to compute the residual Mutual Information between pairs of data vectors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-03-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/134916v1?rss=1">
<title>
<![CDATA[
Activity-Dependent Control Of Inhibitory Interneuron Number In The Mammalian Cortex 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/134916v1?rss=1"
</link>
<description><![CDATA[
Cortical networks are composed of excitatory projection neurons and inhibitory interneurons. Finding the right balance between the two is important for controlling overall cortical excitation and network dynamics. However, it is unclear how the correct number of cortical interneurons (CIs) is established in the mammalian forebrain. CIs are generated in excess from basal forebrain progenitors and their final numbers are adjusted via an intrinsically determined program of apoptosis that takes place during an early postnatal window. Here, we provide evidence that the extent of CI apoptosis during this critical period is plastic, cell type specific and can be reduced in a cell autonomous manner by acute increases in neuronal activity. We propose that the physiological state of the emerging neural network controls the activity levels of local CIs to modulate their numbers in a homeostatic manner.
]]></description>
<dc:creator>Denaxa, M.</dc:creator>
<dc:creator>Neves, G.</dc:creator>
<dc:creator>Rabinowitz, A.</dc:creator>
<dc:creator>Kemlo, S.</dc:creator>
<dc:creator>Liodis, P.</dc:creator>
<dc:creator>Burrone, J.</dc:creator>
<dc:creator>Pachnis, V.</dc:creator>
<dc:date>2017-05-17</dc:date>
<dc:identifier>doi:10.1101/134916</dc:identifier>
<dc:title><![CDATA[Activity-Dependent Control Of Inhibitory Interneuron Number In The Mammalian Cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/123265v1?rss=1">
<title>
<![CDATA[
Pairwise Hybrid Incompatibilities Dominate During Allopatric Speciation For A Simple Genotype-Phenotype Map Of Embryonic Spatial Patterning 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/123265v1?rss=1"
</link>
<description><![CDATA[
Developmental system drift is a likely mechanism for the origin of hybrid incompatibilities between closely related species. We examine here the detailed mechanistic basis of hybrid incompatibilities for a genotype-phenotype map for developmental system drift under stabilising selection, where the organismal phenotype is conserved, but the underlying molecular phenotypes and genotype can drift. This leads to number of emergent phenomenon not obtainable by modelling genotype or phenotype alone. Our results show that: 1) speciation is more rapid at smaller population sizes with a characteristic, Orr-like, power law, but at large population sizes slow, characterised by a sub-diffusive growth law; 2) the molecular phenotypes under weakest selection contribute to the earliest incompatibilities; and 3) pairwise incompatibilities dominate over higher order, contrary to previous predictions that the latter should dominate. Our results indicate that biophysics and population size provide a much stronger constraint to speciation than suggested by previous models.
]]></description>
<dc:creator>Khatri, B. S.</dc:creator>
<dc:creator>Goldstein, R.</dc:creator>
<dc:date>2017-04-02</dc:date>
<dc:identifier>doi:10.1101/123265</dc:identifier>
<dc:title><![CDATA[Pairwise Hybrid Incompatibilities Dominate During Allopatric Speciation For A Simple Genotype-Phenotype Map Of Embryonic Spatial Patterning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/123232v1?rss=1">
<title>
<![CDATA[
Rate Of Fixation Of Rare Variants In A Population 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/123232v1?rss=1"
</link>
<description><![CDATA[
The process of molecular evolution has been dominated by the Kimura paradigm for nearly 60 years; mutations arise at a certain rate in the population and they go to fixation with a probability given by Kimuras classic formula, which assumes there are no further mutations that interfere with the fixation process. An alternative view is that rare variants exist in the population in a mutation-drift-selection balance and rise to fixation through a combination of chance (genetic drift), selection and mutation. When mutations increase in strength, but still in the weak regime, we would expect the Kimura rate approximation to be an overestimate, as a rare variant which grows in frequency will suffer a greater backward flux of mutations, slowing progress to fixation. However, to date calculating important quantities for a general model of selection and mutation, like the rate of fixation of these rare variants has not been tractable in the conventional diffusion approximation of population genetics. Here, we use Fishers angular transformation to convert the frequency-dependent diffusion inherent in population genetics to simple diffusion in an effective potential, which describes the forces of selection, drift and mutation. Once this potential is defined it is simple to show that the mean first passage time is given by a double integral which relate to populations at the barrier. Exact numerical integration shows excellent agreement with discrete Wright-Fisher simulations, which do show a slowing down of the fixation of mutants at higher mutation rates and for strong positive selection, compared to the Kimura prediction. We then seek a closed-form analytical expression for the rate of fixation of mutants, by adapting Kramers approximation for the mean first passage time. This overall gives an accurate approximation, but however, does not improve on the Kimura rate.
]]></description>
<dc:creator>Khatri, B. S.</dc:creator>
<dc:date>2017-04-02</dc:date>
<dc:identifier>doi:10.1101/123232</dc:identifier>
<dc:title><![CDATA[Rate Of Fixation Of Rare Variants In A Population]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/093401v1?rss=1">
<title>
<![CDATA[
Genome sequence of a diabetes-prone desert rodent reveals a mutation hotspot around the ParaHox gene cluster 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/093401v1?rss=1"
</link>
<description><![CDATA[
The sand rat Psammomys obesus is a gerbil native to deserts of North Africa and the Middle East1. Sand rats survive with low caloric intake and when given high carbohydrate diets can become obese and develop type II diabetes2 which, in extreme cases, leads to pancreatic failure and death3,4. Previous studies have reported inability to detect the Pdx1 gene or protein in gerbils5-7, suggesting that absence of this key insulin-regulating homeobox gene might underlie diabetes susceptibility. Here we report sequencing of the sand rat genome and discovery of an extensive, mutationally-biased GC-rich genomic domain encompassing many essential genes, including the elusive Pdx1. The sequence of Pdx1 has been grossly affected by GC-biased mutation leading to the highest divergence observed in the animal kingdom. In addition to molecular insights into restricted caloric intake in a desert species, the discovery that specific chromosomal regions can be subject to elevated mutation rate has widespread significance to evolution.
]]></description>
<dc:creator>Hargreaves, A. D.</dc:creator>
<dc:creator>Zhou, L.</dc:creator>
<dc:creator>Christensen, J.</dc:creator>
<dc:creator>Marletaz, F.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Li, F.</dc:creator>
<dc:creator>Jansen, P. G.</dc:creator>
<dc:creator>Spiga, E.</dc:creator>
<dc:creator>Hansen, M. T.</dc:creator>
<dc:creator>Horn Pedersen, S. V.</dc:creator>
<dc:creator>Biswas, S.</dc:creator>
<dc:creator>Serikawa, K.</dc:creator>
<dc:creator>Fox, B. A.</dc:creator>
<dc:creator>Taylor, W. R.</dc:creator>
<dc:creator>Mulley, J. F.</dc:creator>
<dc:creator>Zhang, G.</dc:creator>
<dc:creator>Heller, R. S.</dc:creator>
<dc:creator>Holland, P. W. H.</dc:creator>
<dc:date>2016-12-17</dc:date>
<dc:identifier>doi:10.1101/093401</dc:identifier>
<dc:title><![CDATA[Genome sequence of a diabetes-prone desert rodent reveals a mutation hotspot around the ParaHox gene cluster]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-12-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/126508v1?rss=1">
<title>
<![CDATA[
Combinatorial Regulation Of The Balance Between Dynein Microtubule End Accumulation And Initiation Of Directed Motility 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/126508v1?rss=1"
</link>
<description><![CDATA[
Cytoplasmic dynein is involved in a multitude of essential cellular functions. Dyneins activity is controlled by the combinatorial action of several regulators. The molecular mechanism of this regulation is poorly understood. Using purified proteins, we reconstitute the regulation of the human dynein complex by three prominent regulators on dynamic microtubules in the presence of end binding proteins (EBs). We find that dynein can be in biochemically and functionally distinct pools: either passively tracking dynamic microtubule plus-ends in an EB-dependent manner or moving processively towards minus ends in an adaptor protein-dependent manner. Whereas both dynein pools share the dynactin complex, they have opposite preferences for binding other regulators, either the adaptor protein Bicaudal D2 (BicD2) or the multifunctional regulator Lisencephaly-1 (Lis1). Remarkably, dynactin, but not EBs, strongly biases motility initiation locally from microtubule plus ends by autonomous plus end recognition. BicD2 and Lis1 together control the overall efficiency of motility initiation. Our study provides insight into the mechanism of dynein activity regulation by dissecting the distinct functional contributions of the individual members of a dynein regulatory network.
]]></description>
<dc:creator>Jha, R.</dc:creator>
<dc:creator>Roostalu, J.</dc:creator>
<dc:creator>Trokter, M.</dc:creator>
<dc:creator>Surrey, T.</dc:creator>
<dc:date>2017-04-11</dc:date>
<dc:identifier>doi:10.1101/126508</dc:identifier>
<dc:title><![CDATA[Combinatorial Regulation Of The Balance Between Dynein Microtubule End Accumulation And Initiation Of Directed Motility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-04-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/051417v1?rss=1">
<title>
<![CDATA[
Mutational signatures associated with tobacco smoking in human cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/051417v1?rss=1"
</link>
<description><![CDATA[
Tobacco smoking increases the risk of at least 15 classes of cancer. We analyzed somatic mutations and DNA methylation in 5,243 cancers of types for which tobacco smoking confers an elevated risk. Smoking is associated with increased mutation burdens of multiple distinct mutational signatures, which contribute to different extents in different cancers. One of these signatures, mainly found in cancers derived from tissues directly exposed to tobacco smoke, is attributable to misreplication of DNA damage caused by tobacco carcinogens. Others likely reflect indirect activation of DNA editing by APOBEC cytidine deaminases and of an endogenous clock-like mutational process. The results are consistent with the proposition that smoking increases cancer risk by increasing the somatic mutation load, although direct evidence for this mechanism is lacking in some smoking-related cancer types.nnONE SENTENCE SUMMARYMultiple distinct mutational processes associate with tobacco smoking in cancer reflecting direct and indirect effects of tobacco smoke.
]]></description>
<dc:creator>Ludmil B Alexandrov</dc:creator>
<dc:creator>Young Seok Ju</dc:creator>
<dc:creator>Kerstin Haase</dc:creator>
<dc:creator>Peter Van Loo</dc:creator>
<dc:creator>Inigo Martincorena</dc:creator>
<dc:creator>Serena Nik-Zainal</dc:creator>
<dc:creator>Yasushi Totoki</dc:creator>
<dc:creator>Akihiro Fujimoto</dc:creator>
<dc:creator>Hidewaki Nakagawa</dc:creator>
<dc:creator>Tatsuhiro Shibata</dc:creator>
<dc:creator>Peter J Campbell</dc:creator>
<dc:creator>Paolo Vineis</dc:creator>
<dc:creator>David H Phillips</dc:creator>
<dc:creator>Michael R Stratton</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-05-03</dc:date>
<dc:identifier>doi:10.1101/051417</dc:identifier>
<dc:title><![CDATA[Mutational signatures associated with tobacco smoking in human cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-05-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/132324v1?rss=1">
<title>
<![CDATA[
Universal Patterns Of Selection In Cancer And Somatic Tissues 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/132324v1?rss=1"
</link>
<description><![CDATA[
Cancer develops as a result of somatic mutation and clonal selection, but quantitative measures of selection in cancer evolution are lacking. We applied methods from evolutionary genomics to 7,664 human cancers across 29 tumor types. Unlike species evolution, positive selection outweighs negative selection during cancer development. On average, <1 coding base substitution/tumor is lost through negative selection, with purifying selection only detected for truncating mutations in essential genes in haploid regions. This allows exome-wide enumeration of all driver mutations, including outside known cancer genes. On average, tumors carry [~]4 coding substitutions under positive selection, ranging from <1/tumor in thyroid and testicular cancers to >10/tumor in endometrial and colorectal cancers. Half of driver substitutions occur in yet-to-be-discovered cancer genes. With increasing mutation burden, numbers of driver mutations increase, but not linearly. We identify novel cancer genes and show that genes vary extensively in what proportion of mutations are drivers versus passengers.nnHIGHLIGHTSO_LIUnlike the germline, somatic cells evolve predominantly by positive selectionnC_LIO_LINearly all ([~]99%) coding mutations are tolerated and escape negative selectionnC_LIO_LIFirst exome-wide estimates of the total number of driver coding mutations per tumornC_LIO_LI1-10 coding driver mutations per tumor; half occurring outside known cancer genesnC_LI
]]></description>
<dc:creator>Martincorena, I.</dc:creator>
<dc:creator>Raine, K. M.</dc:creator>
<dc:creator>Gerstung, M.</dc:creator>
<dc:creator>Dawson, K. J.</dc:creator>
<dc:creator>Haase, K.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>Davies, H.</dc:creator>
<dc:creator>Stratton, M. R.</dc:creator>
<dc:creator>Campbell, P. J.</dc:creator>
<dc:date>2017-04-29</dc:date>
<dc:identifier>doi:10.1101/132324</dc:identifier>
<dc:title><![CDATA[Universal Patterns Of Selection In Cancer And Somatic Tissues]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-04-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/023317v1?rss=1">
<title>
<![CDATA[
GOTHiC, a simple probabilistic model to resolve complex biases and to identify real interactions in Hi-C data 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/023317v1?rss=1"
</link>
<description><![CDATA[
1.Hi-C is one of the main methods for investigating spatial co-localisation of DNA in the nucleus. However, the raw sequencing data obtained from Hi-C experiments suffer from large biases and spurious contacts, making it difficult to identify true interactions. Existing methods use complex models to account for biases and do not provide a significance threshold for detecting interactions. Here we introduce a simple binomial probabilistic model that resolves complex biases and distinguishes between true and false interactions. The model corrects biases of known and unknown origin and yields a p-value for each interaction, providing a reliable threshold based on significance. We demonstrate this experimentally by testing the method against a random ligation dataset. Our method outperforms previous methods and provides a statistical framework for further data analysis, such as comparisons of Hi-C interactions between different conditions. GOTHiC is available as a BioConductor package (http://www.bioconductor.org/packages/release/bioc/html/GOTHiC.html).
]]></description>
<dc:creator>Borbala Mifsud</dc:creator>
<dc:creator>Inigo Martincorena</dc:creator>
<dc:creator>Elodie Darbo</dc:creator>
<dc:creator>Robert Sugar</dc:creator>
<dc:creator>Stefan Schoenfelder</dc:creator>
<dc:creator>Peter Fraser</dc:creator>
<dc:creator>Nicholas Luscombe</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-07-27</dc:date>
<dc:identifier>doi:10.1101/023317</dc:identifier>
<dc:title><![CDATA[GOTHiC, a simple probabilistic model to resolve complex biases and to identify real interactions in Hi-C data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-07-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/117291v1?rss=1">
<title>
<![CDATA[
Evolution and clinical impact of genetic epistasis within EGFR-mutant lung cancers 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/117291v1?rss=1"
</link>
<description><![CDATA[
Introductory paragraph Introductory paragraph Main text Methods Author Contributions Author Information References The current understanding of tumorigenesis is largely centered on a monogenic driver oncogene model. This paradigm is incompatible with the prevailing clinical experience in most solid malignancies: monotherapy with a drug directed against an individual oncogenic driver typically results in incomplete clinical responses and eventual tumor progression1-7. By profiling the somatic genetic alterations present in over 2,000 cases of lung cancer, the leading cause of cancer mortality worldwide8,9, we show that combinations of functional genetic alterations, i.e. genetic collectives dominate the landscape of ...
]]></description>
<dc:creator>Blakely, C. M.</dc:creator>
<dc:creator>Watkins, T. B. K.</dc:creator>
<dc:creator>Wu, W.</dc:creator>
<dc:creator>Gini, B.</dc:creator>
<dc:creator>Chabon, J. J.</dc:creator>
<dc:creator>McCoach, C. E.</dc:creator>
<dc:creator>McGranahan, N.</dc:creator>
<dc:creator>Wilson, G. A.</dc:creator>
<dc:creator>Birkbak, N. J.</dc:creator>
<dc:creator>Olivas, V.</dc:creator>
<dc:creator>Rotow, J.</dc:creator>
<dc:creator>Maynard, A.</dc:creator>
<dc:creator>Wang, V.</dc:creator>
<dc:creator>Gubens, M.</dc:creator>
<dc:creator>Banks, K.</dc:creator>
<dc:creator>Lanman, R.</dc:creator>
<dc:creator>Caulin, A.</dc:creator>
<dc:creator>St. John, J.</dc:creator>
<dc:creator>Cordero, A.</dc:creator>
<dc:creator>Giannikopoulos, P.</dc:creator>
<dc:creator>Mack, P.</dc:creator>
<dc:creator>Gandara, D.</dc:creator>
<dc:creator>Husain, H.</dc:creator>
<dc:creator>Doebele, R.</dc:creator>
<dc:creator>Riess, J.</dc:creator>
<dc:creator>Diehn, M.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Bivona, T. G.</dc:creator>
<dc:date>2017-03-16</dc:date>
<dc:identifier>doi:10.1101/117291</dc:identifier>
<dc:title><![CDATA[Evolution and clinical impact of genetic epistasis within EGFR-mutant lung cancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-03-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/073478v1?rss=1">
<title>
<![CDATA[
Precise label-free quantitative proteomes in high-throughput by microLC and data-independent SWATH acquisition 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/073478v1?rss=1"
</link>
<description><![CDATA[
While quantitative proteomics is a key technology in biological research, the routine industry and diagnostics application is so far still limited by a moderate throughput, data consistency and robustness. In part, the restrictions emerge in the proteomics dependency on nanolitre/minute flow rate chromatography that enables a high sensitivity, but is difficult to handle on large sample series, and on the stochastic nature in data-dependent acquisition strategies. We here establish and benchmark a label-free, quantitative proteomics platform that uses microlitre/minute flow rate chromatography in combination with data-independent SWATH acquisition. Being able to largely compensate for the loss of sensitivity by exploiting the analytical capacities of microflow chromatography, we show that microLC-SWATH-MS is able to precisely quantify up to 4000 proteins in an hour or less, enables the consistent processing of sample series in high-throughput, and gains quantification precisions comparable to targeted proteomic assays. MicroLC-SWATH-MS can hence routinely process hundreds to thousands of samples to systematically create precise, label free quantitative proteomes.
]]></description>
<dc:creator>Jakob Vowinckel</dc:creator>
<dc:creator>Aleksej Zelezniak</dc:creator>
<dc:creator>Artur Kibler</dc:creator>
<dc:creator>Roland Bruderer</dc:creator>
<dc:creator>Michael Muelleder</dc:creator>
<dc:creator>Lukas Reiter</dc:creator>
<dc:creator>Markus Ralser</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-09-05</dc:date>
<dc:identifier>doi:10.1101/073478</dc:identifier>
<dc:title><![CDATA[Precise label-free quantitative proteomes in high-throughput by microLC and data-independent SWATH acquisition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/106971v1?rss=1">
<title>
<![CDATA[
FAM83G/PAWS1 controls cytoskeletal dynamics and cell migration through association with the SH3 adaptor CD2AP 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/106971v1?rss=1"
</link>
<description><![CDATA[
Our previous studies of PAWS1 (Protein Associated With SMAD1) have suggested that this molecule has roles beyond BMP signalling. To investigate these roles, we have used CRISPR/Cas9 to generate PAWS1 knockout cells. Here, we show that PAWS1 plays a role in the regulation of the cytoskeletal machinery, including actin and focal adhesion dynamics, and cell migration. Confocal microscopy and live cell imaging of actin in U2OS cells indicate that PAWS1 is also involved in cytoskeletal dynamics and organization. Loss of PAWS1 causes severe defects in F-actin organization and distribution as well as in lamellipodial organization, resulting in impaired cell migration. PAWS1 interacts in a dynamic fashion with the actin/cytoskeletal regulator CD2AP at lamellae, suggesting that its association with CD2AP controls actin organization and cellular migration.nnSummary statementPAWS1/FAM83G controls cell migration by influencing the organisation of F-actin and focal adhesions and the distribution of the actin stress fibre network through its association with CD2AP.
]]></description>
<dc:creator>Sapkota, G. P.</dc:creator>
<dc:creator>Cummins, T. D.</dc:creator>
<dc:creator>Wu, K. Z. L.</dc:creator>
<dc:creator>Bozatzi, P.</dc:creator>
<dc:creator>Dingwell, K. S.</dc:creator>
<dc:creator>Macartney, T. J.</dc:creator>
<dc:creator>Wood, N. T.</dc:creator>
<dc:creator>Varghese, J.</dc:creator>
<dc:creator>Gourlay, R.</dc:creator>
<dc:creator>Campbell, D. G.</dc:creator>
<dc:creator>Prescott, A.</dc:creator>
<dc:creator>Griffis, E.</dc:creator>
<dc:creator>Smith, J. C.</dc:creator>
<dc:date>2017-02-08</dc:date>
<dc:identifier>doi:10.1101/106971</dc:identifier>
<dc:title><![CDATA[FAM83G/PAWS1 controls cytoskeletal dynamics and cell migration through association with the SH3 adaptor CD2AP]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-02-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/104307v1?rss=1">
<title>
<![CDATA[
Olig2 and Hes regulatory dynamics during motor neuron differentiation revealed by single cell transcriptomics 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/104307v1?rss=1"
</link>
<description><![CDATA[
During tissue development, multipotent progenitors differentiate into specific cell types in characteristic spatial and temporal patterns. We address the mechanism linking progenitor identity and differentiation rate in the neural tube, where motor neuron (MN) progenitors differentiate more rapidly than other progenitors. Using single cell transcriptomics, we define the transcriptional changes associated with the transition of neural progenitors into MNs. Reconstruction of gene expression dynamics from these data indicate a pivotal role for the MN determinant Olig2 just prior to MN differentiation. Olig2 represses expression of the Notch signaling pathway effectors Hes1 and Hes5. Olig2 repression of Hes5 appears to be direct, via a conserved regulatory element within the Hes5 locus that restricts expression from MN progenitors. These findings reveal a tight coupling between the regulatory networks that control patterning and neuronal differentiation, and demonstrate how Olig2 acts as the developmental pacemaker coordinating the spatial and temporal pattern of MN generation.
]]></description>
<dc:creator>Sagner, A.</dc:creator>
<dc:creator>Gaber, Z.</dc:creator>
<dc:creator>Delile, J.</dc:creator>
<dc:creator>Kong, J. H.</dc:creator>
<dc:creator>Rousso, D. L.</dc:creator>
<dc:creator>Pearson, C. A.</dc:creator>
<dc:creator>Weicksel, S. E.</dc:creator>
<dc:creator>Mousavy Gharavy, N.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Novitch, B.</dc:creator>
<dc:date>2017-01-30</dc:date>
<dc:identifier>doi:10.1101/104307</dc:identifier>
<dc:title><![CDATA[Olig2 and Hes regulatory dynamics during motor neuron differentiation revealed by single cell transcriptomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-01-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/152637v1?rss=1">
<title>
<![CDATA[
Epidermal Wnt signalling regulates transcriptome heterogeneity and proliferative fate in neighbouring cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/152637v1?rss=1"
</link>
<description><![CDATA[
Canonical Wnt/beta-catenin signalling regulates self-renewal and lineage selection within the mouse epidermis. Although the transcriptional response of keratinocytes that receive a Wnt signal is well characterised, little is known about the mechanism by which keratinocytes in proximity to the Wntreceiving cell are co-opted to undergo a change in cell fate. To address this, we performed single-cell mRNA-Seq on mouse keratinocytes co-cultured with and without the presence of beta-catenin activated neighbouring cells. We identified seven distinct cell states in cultures that had not been exposed to the beta-catenin stimulus and show that the stimulus redistributes wild type subpopulation proportions. Using temporal single-cell analysis we reconstruct the cell fate changes induced by neighbour Wnt activation. Gene expression heterogeneity was reduced in neighbouring cells and this effect was most dramatic for protein synthesis associated genes. The changes in gene expression were accompanied by a shift from a quiescent to a more proliferative stem cell state. By integrating imaging and reconstructed sequential gene expression changes during the state transition we identified transcription factors, including Smad4 and Bcl3, that were responsible for effecting the transition in a contact-dependent manner. Our data indicate that non cell-autonomous Wnt/beta-catenin signalling decreases transcriptional heterogeneity and further our understanding of how epidermal Wnt signalling orchestrates regeneration and self-renewal.
]]></description>
<dc:creator>Ghahramani, A.</dc:creator>
<dc:creator>Donati, G.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:creator>Watt, F. M.</dc:creator>
<dc:date>2017-06-20</dc:date>
<dc:identifier>doi:10.1101/152637</dc:identifier>
<dc:title><![CDATA[Epidermal Wnt signalling regulates transcriptome heterogeneity and proliferative fate in neighbouring cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/158006v1?rss=1">
<title>
<![CDATA[
Neutral tumor evolution? 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/158006v1?rss=1"
</link>
<description><![CDATA[
Tumor growth is an evolutionary process governed by somatic mutation, clonal selection and random genetic drift, constrained by the co-evolution of the microenvironment1,2. Tumor subclones are subpopulations of tumor cells with a common set of mutations resulting from the expansion of a single cell during tumor development, and have been observed in a significant fraction of cancers and across multiple cancer types3. Peter Nowell proposed that tumors evolve through sequential genetic events4, whereby one cell acquires a selective advantage so that its lineage becomes predominant. According to this traditional model, the selective advantage is conferred by a small set of driver mutations, but, as the subclones that bear them expand successively, they accumulate passenger mutations as well, which can be detected in sequencing experiments1. Genomes of indiv ...
]]></description>
<dc:creator>Tarabichi, M.</dc:creator>
<dc:creator>Martincorena, I.</dc:creator>
<dc:creator>Gerstung, M.</dc:creator>
<dc:creator>Markowetz, F.</dc:creator>
<dc:creator>Spellman, P. T.</dc:creator>
<dc:creator>Morris, Q. D.</dc:creator>
<dc:creator>Lingjaerde, O. C.</dc:creator>
<dc:creator>Wedge, D. C.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>- PCAWG Evolution and Heterogeneity Working Group,</dc:creator>
<dc:date>2017-06-30</dc:date>
<dc:identifier>doi:10.1101/158006</dc:identifier>
<dc:title><![CDATA[Neutral tumor evolution?]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/166017v1?rss=1">
<title>
<![CDATA[
Allele-specific multi-sample copy number segmentation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/166017v1?rss=1"
</link>
<description><![CDATA[
MotivationAllele-specific copy number alterations are commonly used to trace the evolution of tumours. A key step of the analysis is to segment genomic data into regions of constant copy number. For precise phylogenetic inference, breakpoints shared between samples need to be aligned to each other.nnResultsHere we present asmultipcf, an algorithm for allele-specific segmentation of multiple samples that infers private and shared segment boundaries of phylogenetically related samples. The output of this algorithm can directly be used for allele-specific copy number calling using ASCAT.nnAvailabilityasmultipcf is available as part of the ASCAT R package (version 2.5) from github.com/Crick-CancerGenomics/ascat
]]></description>
<dc:creator>Ross, E. M.</dc:creator>
<dc:creator>Haase, K.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>Markowetz, F.</dc:creator>
<dc:date>2017-07-20</dc:date>
<dc:identifier>doi:10.1101/166017</dc:identifier>
<dc:title><![CDATA[Allele-specific multi-sample copy number segmentation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/168153v1?rss=1">
<title>
<![CDATA[
Genomic landscape of oxidative DNA damage and repair reveals regioselective protection from mutagenesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/168153v1?rss=1"
</link>
<description><![CDATA[
DNA is subject to constant chemical modification and damage, which eventually results in variable mutation rates throughout the genome. Although detailed molecular mechanisms of DNA damage and repair are well-understood, damage impact and execution of repair across a genome remains poorly defined. To bridge the gap between our understanding of DNA repair and mutation distributions we developed a novel method, AP-seq, capable of mapping apurinic sitesand 8-oxo-7,8-dihydroguanine bases at [~]300bp resolution on a genome-wide scale. We directly demonstrate that the accumulation rate of oxidative damage varies widely across the genome, with hot spots acquiring many times more damage than cold spots. Unlike SNVs in cancers, damage burden correlates with marks for open chromatin notably H3K9ac and H3K4me2. Oxidative damage is also highly enriched in transposable elements and other repetitive sequences. In contrast, we observe decreased damage at promoters, exons and termination sites, but not introns, in a seemingly transcription-independent manner. Leveraging cancer genomic data, we also find locally reduced SNV rates in promoters, genes and other functional elements. Taken together, our study reveals that oxidative DNA damage accumulation and repair differ strongly across the genome, but culminate in a previously unappreciated mechanism that safe-guards the regulatory sequences and the coding regions of genes from mutations.
]]></description>
<dc:creator>Poetsch, A. R.</dc:creator>
<dc:creator>Boulton, S. J.</dc:creator>
<dc:creator>Luscombe, N.</dc:creator>
<dc:date>2017-07-25</dc:date>
<dc:identifier>doi:10.1101/168153</dc:identifier>
<dc:title><![CDATA[Genomic landscape of oxidative DNA damage and repair reveals regioselective protection from mutagenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/170100v1?rss=1">
<title>
<![CDATA[
3’UTR Remodelling of Axonal Transcripts in Sympathetic Neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/170100v1?rss=1"
</link>
<description><![CDATA[
The 3 untranslated regions (3UTRs) of messenger RNAs (mRNA) are non-coding sequences that regulate several aspects of mRNA metabolism, including intracellular localisation and translation. Here, we show that in sympathetic neuron axons, the 3UTRs of many transcripts undergo cleavage, generating both translatable isoforms expressing a shorter 3UTR, and 3UTR fragments. 3end RNA sequencing indicated that 3UTR cleavage is a potentially widespread event in axons, which is mediated by a protein complex containing the endonuclease Ago2 and the RNA binding protein HuD. Analysis of the Inositol monophosphatase 1 (Impa1) mRNA revealed that a stem loop structure within the 3UTR is necessary for Ago2 cleavage. Thus, remodeling of the 3UTR provides an alternative mechanism that simultaneously regulates local protein synthesis and generates a new class of 3UTR RNAs with yet unknown functions.
]]></description>
<dc:creator>Andreassi, C.</dc:creator>
<dc:creator>Luisier, R.</dc:creator>
<dc:creator>Crerar, H.</dc:creator>
<dc:creator>Franke, S.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:creator>Cuda, G.</dc:creator>
<dc:creator>Gaspari, M.</dc:creator>
<dc:creator>Riccio, A.</dc:creator>
<dc:date>2017-07-30</dc:date>
<dc:identifier>doi:10.1101/170100</dc:identifier>
<dc:title><![CDATA[3’UTR Remodelling of Axonal Transcripts in Sympathetic Neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/161562v1?rss=1">
<title>
<![CDATA[
The evolutionary history of 2,658 cancers 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/161562v1?rss=1"
</link>
<description><![CDATA[
Cancer develops through a process of somatic evolution. Here, we use whole-genome sequencing of 2,778 tumour samples from 2,658 donors to reconstruct the life history, evolution of mutational processes, and driver mutation sequences of 39 cancer types. The early phases of oncogenesis are driven by point mutations in a small set of driver genes, often including biallelic inactivation of tumour suppressors. Early oncogenesis is also characterised by specific copy number gains, such as trisomy 7 in glioblastoma or isochromosome 17q in medulloblastoma. By contrast, increased genomic instability, a nearly four-fold diversification of driver genes, and an acceleration of point mutation processes are features of later stages. Copy-number alterations often occur in mitotic crises leading to simultaneous gains of multiple chromosomal segments. Timing analysis suggests that driver mutations often precede diagnosis by many years, and in some cases decades, providing a window of opportunity for early cancer detection.
]]></description>
<dc:creator>Gerstung, M.</dc:creator>
<dc:creator>Jolly, C.</dc:creator>
<dc:creator>Leshchiner, I.</dc:creator>
<dc:creator>Dentro, S. C.</dc:creator>
<dc:creator>Gonzalez, S.</dc:creator>
<dc:creator>Mitchell, T. J.</dc:creator>
<dc:creator>Rubanova, Y.</dc:creator>
<dc:creator>Anur, P.</dc:creator>
<dc:creator>Rosebrock, D.</dc:creator>
<dc:creator>Yu, K.</dc:creator>
<dc:creator>Tarabichi, M.</dc:creator>
<dc:creator>Deshwar, A.</dc:creator>
<dc:creator>Wintersinger, J.</dc:creator>
<dc:creator>Kleinheinz, K.</dc:creator>
<dc:creator>Vazquez-Garcia, I.</dc:creator>
<dc:creator>Haase, K.</dc:creator>
<dc:creator>Sengupta, S.</dc:creator>
<dc:creator>Macintyre, G.</dc:creator>
<dc:creator>Malikic, S.</dc:creator>
<dc:creator>Donmez, N.</dc:creator>
<dc:creator>Livitz, D. G.</dc:creator>
<dc:creator>Cmero, M.</dc:creator>
<dc:creator>Demeulemeester, J.</dc:creator>
<dc:creator>Schumacher, S.</dc:creator>
<dc:creator>Fan, Y.</dc:creator>
<dc:creator>Yao, X.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Schlesner, M.</dc:creator>
<dc:creator>Boutros, P. C.</dc:creator>
<dc:creator>Bowtell, D. D.</dc:creator>
<dc:creator>Zhu, H.</dc:creator>
<dc:creator>Getz, G.</dc:creator>
<dc:creator>Imielinski, M.</dc:creator>
<dc:creator>Beroukhim, R.</dc:creator>
<dc:creator>Sahinalp, S. C.</dc:creator>
<dc:creator>Ji, Y.</dc:creator>
<dc:creator>Peifer, M.</dc:creator>
<dc:creator>Markowetz, F.</dc:creator>
<dc:creator>Mustonen, V.</dc:creator>
<dc:creator>Yuan, K.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Morris, Q. D.</dc:creator>
<dc:creator>Spellman, P. T.</dc:creator>
<dc:creator>Wedge, D. C.</dc:creator>
<dc:creator>V</dc:creator>
<dc:date>2017-07-11</dc:date>
<dc:identifier>doi:10.1101/161562</dc:identifier>
<dc:title><![CDATA[The evolutionary history of 2,658 cancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/167155v1?rss=1">
<title>
<![CDATA[
Probing tissue-scale deformation by in vivo force application reveals a fast tissue softening during early embryogenesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/167155v1?rss=1"
</link>
<description><![CDATA[
During development, cell-generated forces induce tissue-scale deformations to shape the organism. Here, we present a method that allows to quantitatively relate such tissue-scale deformations to spatially localized forces and measure mechanical properties of epithelia in vivo. Our approach is based on the application of controlled forces on microparticles embedded in individual cells of an embryo. Combining measurements of the bead displacement with the analysis of induced deformation fields in a continuum mechanics framework, we can quantify tissue material properties and follow their change over time. In particular, we uncover a rapid change in tissue response occurring during Drosophila cellularization, resulting from a softening of the blastoderm and an increase of external friction. Pharmacological treatments reveal that in addition to actomyosin, the microtubule cytoskeleton is a major contributor to epithelial mechanics at that stage. Overall, our method allows for measuring essential mechanical parameters governing tissue-scale deformations and flows occurring during morphogenesis.
]]></description>
<dc:creator>D'Angelo, A.</dc:creator>
<dc:creator>Dierkes, K.</dc:creator>
<dc:creator>Carolis, C.</dc:creator>
<dc:creator>Salbreux, G.</dc:creator>
<dc:creator>Solon, J.</dc:creator>
<dc:date>2017-07-21</dc:date>
<dc:identifier>doi:10.1101/167155</dc:identifier>
<dc:title><![CDATA[Probing tissue-scale deformation by in vivo force application reveals a fast tissue softening during early embryogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/161638v1?rss=1">
<title>
<![CDATA[
Large-Scale Uniform Analysis of Cancer Whole Genomes in Multiple Computing Environments 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/161638v1?rss=1"
</link>
<description><![CDATA[
The International Cancer Genome Consortium (ICGC)s Pan-Cancer Analysis of Whole Genomes (PCAWG) project aimed to categorize somatic and germline variations in both coding and non-coding regions in over 2,800 cancer patients. To provide this dataset to the research working groups for downstream analysis, the PCAWG Technical Working Group marshalled ~800TB of sequencing data from distributed geographical locations; developed portable software for uniform alignment, variant calling, artifact filtering and variant merging; performed the analysis in a geographically and technologically disparate collection of compute environments; and disseminated high-quality validated consensus variants to the working groups. The PCAWG dataset has been mirrored to multiple repositories and can be located using the ICGC Data Portal. The PCAWG workflows are also available as Docker images through Dockstore enabling researchers to replicate our analysis on their own data.
]]></description>
<dc:creator>Yung, C. K.</dc:creator>
<dc:creator>O'Connor, B. D.</dc:creator>
<dc:creator>Yakneen, S.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Ellrott, K.</dc:creator>
<dc:creator>Kleinheinz, K.</dc:creator>
<dc:creator>Miyoshi, N.</dc:creator>
<dc:creator>Raine, K. M.</dc:creator>
<dc:creator>Royo, R.</dc:creator>
<dc:creator>Saksena, G. B.</dc:creator>
<dc:creator>Schlesner, M.</dc:creator>
<dc:creator>Shorser, S. I.</dc:creator>
<dc:creator>Vazquez, M.</dc:creator>
<dc:creator>Weischenfeldt, J.</dc:creator>
<dc:creator>Yuen, D.</dc:creator>
<dc:creator>Butler, A. P.</dc:creator>
<dc:creator>Davis-Dusenbery, B. N.</dc:creator>
<dc:creator>Eils, R.</dc:creator>
<dc:creator>Ferretti, V.</dc:creator>
<dc:creator>Grossman, R. L.</dc:creator>
<dc:creator>Harismendy, O.</dc:creator>
<dc:creator>Kim, Y.</dc:creator>
<dc:creator>Nakagawa, H.</dc:creator>
<dc:creator>Newhouse, S. J.</dc:creator>
<dc:creator>Torrents, D.</dc:creator>
<dc:creator>Stein, L. D.</dc:creator>
<dc:creator>- PCAWG Technical Working Group,</dc:creator>
<dc:date>2017-07-10</dc:date>
<dc:identifier>doi:10.1101/161638</dc:identifier>
<dc:title><![CDATA[Large-Scale Uniform Analysis of Cancer Whole Genomes in Multiple Computing Environments]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/179705v1?rss=1">
<title>
<![CDATA[
Pan-cancer analysis of whole genomes reveals driver rearrangements promoted by LINE-1 retrotransposition in human tumours 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/179705v1?rss=1"
</link>
<description><![CDATA[
About half of all cancers have somatic integrations of retrotransposons. To characterize their role in oncogenesis, we analyzed the patterns and mechanisms of somatic retrotransposition in 2,954 cancer genomes from 37 histological cancer subtypes. We identified 19,166 somatically acquired retrotransposition events, affecting 35% of samples, and spanning a range of event types. L1 insertions emerged as the first most frequent type of somatic structural variation in esophageal adenocarcinoma, and the second most frequent in head-and-neck and colorectal cancers. Aberrant L1 integrations can delete megabase-scale regions of a chromosome, sometimes removing tumour suppressor genes, as well as inducing complex translocations and large-scale duplications. Somatic retrotranspositions can also initiate breakage-fusion-bridge cycles, leading to high-level amplification of oncogenes. These observations illuminate a relevant role of L1 retrotransposition in remodeling the cancer genome, with potential implications in the development of human tumours.
]]></description>
<dc:creator>Rodriguez-Martin, B.</dc:creator>
<dc:creator>Alvarez, E. G.</dc:creator>
<dc:creator>Baez-Ortega, A.</dc:creator>
<dc:creator>Demeulemeester, J.</dc:creator>
<dc:creator>Ju, Y. S.</dc:creator>
<dc:creator>Zamora, J.</dc:creator>
<dc:creator>Detering, H.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Contino, G.</dc:creator>
<dc:creator>Dentro, S. C.</dc:creator>
<dc:creator>Bruzos, A. L.</dc:creator>
<dc:creator>Dueso-Barroso, A.</dc:creator>
<dc:creator>Ardeljan, D.</dc:creator>
<dc:creator>Tojo, M.</dc:creator>
<dc:creator>Roberts, N. D.</dc:creator>
<dc:creator>Blanco, M. G.</dc:creator>
<dc:creator>Edwards, P. A. W.</dc:creator>
<dc:creator>Weischenfeldt, J.</dc:creator>
<dc:creator>Santamarina, M.</dc:creator>
<dc:creator>Puiggros, M.</dc:creator>
<dc:creator>Chong, Z.</dc:creator>
<dc:creator>Chen, K.</dc:creator>
<dc:creator>Lee, E. A.</dc:creator>
<dc:creator>Wala, J. A.</dc:creator>
<dc:creator>Raine, K.</dc:creator>
<dc:creator>Butler, A.</dc:creator>
<dc:creator>Waszak, S. M.</dc:creator>
<dc:creator>Navarro, F. C. P.</dc:creator>
<dc:creator>Schumacher, S. E.</dc:creator>
<dc:creator>Monlong, J.</dc:creator>
<dc:creator>Maura, F.</dc:creator>
<dc:creator>Bolli, N.</dc:creator>
<dc:creator>Bourque, G.</dc:creator>
<dc:creator>Gerstein, M.</dc:creator>
<dc:creator>Park, P. J.</dc:creator>
<dc:creator>Berroukhim, R.</dc:creator>
<dc:creator>Torrents, D.</dc:creator>
<dc:creator>Korbel, J. O.</dc:creator>
<dc:creator>Martincorena, I.</dc:creator>
<dc:creator>Fitzgerald, R. C.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>Ka</dc:creator>
<dc:date>2017-08-24</dc:date>
<dc:identifier>doi:10.1101/179705</dc:identifier>
<dc:title><![CDATA[Pan-cancer analysis of whole genomes reveals driver rearrangements promoted by LINE-1 retrotransposition in human tumours]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-08-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/180372v1?rss=1">
<title>
<![CDATA[
Post-transcriptional remodelling is temporally deregulated during motor neurogenesis in human ALS models 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/180372v1?rss=1"
</link>
<description><![CDATA[
Mutations causing amyotrophic lateral sclerosis (ALS) strongly implicate regulators of RNA-processing that are ubiquitously expressed throughout development. To understand the molecular impact of ALS-causing mutations on early neuronal development and disease, we performed transcriptomic analysis of differentiated human control and VCP-mutant induced pluripotent stem cells (iPSCs) during motor neurogenesis. We identify intron retention (IR) as the predominant splicing change affecting early stages of wild-type neural differentiation, targeting key genes involved in the splicing machinery. Importantly, IR occurs prematurely in VCP-mutant cultures compared with control counterparts; these events are also observed in independent RNAseq datasets from SOD1- and FUS-mutant motor neurons (MNs). Together with related effects on 3UTR length variation, these findings implicate alternative RNA-processing in regulating distinct stages of lineage restriction from iPSCs to MNs, and reveal a temporal deregulation of such processing by ALS mutations. Thus, ALS-causing mutations perturb the same post-transcriptional mechanisms that underlie human motor neurogenesis.nnHIGHLIGHTSO_LIIntron retention is the main mode of alternative splicing in early differentiation.nC_LIO_LIThe ALS-causing VCP mutation leads to premature intron retention.nC_LIO_LIIncreased intron retention is seen with multiple ALS-causing mutations.nC_LIO_LITranscriptional programs are unperturbed despite post-transcriptional defects.nC_LInneTOC BLURBLuisier et al. identify post-transcriptional changes underlying human motor neurogenesis: extensive variation in 3 UTR length and intron retention (IR) are the early predominant modes of splicing. The VCP mutation causes IR to occur prematurely during motor neurogenesis and these events are validated in other ALS-causing mutations, SOD1 and FUS.
]]></description>
<dc:creator>Luisier, R.</dc:creator>
<dc:creator>Tyzack, G. E.</dc:creator>
<dc:creator>Hall, C. E.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:creator>Patani, R.</dc:creator>
<dc:date>2017-08-24</dc:date>
<dc:identifier>doi:10.1101/180372</dc:identifier>
<dc:title><![CDATA[Post-transcriptional remodelling is temporally deregulated during motor neurogenesis in human ALS models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-08-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/189050v1?rss=1">
<title>
<![CDATA[
Systematic mapping of the free energy landscapes of a growing immunoglobulin domain identifies a kinetic intermediate associated with co-translational proline isomerization 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/189050v1?rss=1"
</link>
<description><![CDATA[
Co-translational folding is a fundamental molecular process that ensures efficient protein biosynthesis and minimizes the wasteful or hazardous formation of misfolded states. However, the complexity of this process makes it extremely challenging to obtain structural characterizations of co-translational folding pathways. Here we contrast observations in translationally-arrested nascent chains with those of a systematic C-terminal truncation strategy. We create a detailed description of chain length-dependent free energy landscapes associated with folding of the FLN5 filamin domain, in isolation and on the ribosome. By using this approach we identify and characterize two folding intermediates, including a partially folded intermediate associated with the isomerization of a conserved proline residue, which, together with measurements of folding kinetics, raises the prospect that neighboring unfolded domains might accumulate during biosynthesis. We develop a simple model to quantify the risk of misfolding in this situation, and show that catalysis of folding by peptidyl-prolyl isomerases is essential to eliminate this hazard.
]]></description>
<dc:creator>Waudby, C. A.</dc:creator>
<dc:creator>Karyadi, M.-E.</dc:creator>
<dc:creator>Wlodarski, T.</dc:creator>
<dc:creator>Cassaignau, A. M. E.</dc:creator>
<dc:creator>Chan, S.</dc:creator>
<dc:creator>Schmidt-Engler, J. M.</dc:creator>
<dc:creator>Wentink, A. S.</dc:creator>
<dc:creator>Camilloni, C.</dc:creator>
<dc:creator>Vendruscolo, M.</dc:creator>
<dc:creator>Cabrita, L. D.</dc:creator>
<dc:creator>Christodoulou, J.</dc:creator>
<dc:date>2017-09-14</dc:date>
<dc:identifier>doi:10.1101/189050</dc:identifier>
<dc:title><![CDATA[Systematic mapping of the free energy landscapes of a growing immunoglobulin domain identifies a kinetic intermediate associated with co-translational proline isomerization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-09-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/172106v1?rss=1">
<title>
<![CDATA[
ATR is a multifunctional regulator of male mouse meiosis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/172106v1?rss=1"
</link>
<description><![CDATA[
Meiotic cells undergo genetic exchange between homologous chromosomes through programmed DNA double-strand break (DSB) formation, recombination and synapsis1, 2. In mice, the DNA damage-regulated phosphatidylinositol-3-kinase-like kinase (PIKK) ATM regulates all of these processes3-6. However, the meiotic functions of another major PIKK, ATR, have remained elusive, because germ line-specific depletion of this kinase is challenging. Using an efficient conditional strategy, we uncover roles for ATR in male mouse prophase I progression. Deletion of ATR causes chromosome axis fragmentation and germ cell elimination at mid pachynema. ATR is required for homologous synapsis, in a manner genetically dissociable from DSB formation. In addition, ATR regulates loading of recombinases RAD51 and DMC1 to DSBs and maintenance of recombination foci on synapsed and asynapsed chromosomes. Mid pachytene spermatocyte elimination in ATR deficient mice cannot be rescued by deletion of ATM and the third DNA damage-regulated PIKK, PRKDC, consistent with the existence of a PIKK-independent surveillance mechanism in the mammalian germ line. Our studies identify ATR as a multifunctional regulator of mouse meiosis.
]]></description>
<dc:creator>Widger, A.</dc:creator>
<dc:creator>Mahadevaiah, S. K.</dc:creator>
<dc:creator>Lange, J.</dc:creator>
<dc:creator>ElInati, E.</dc:creator>
<dc:creator>Zohren, J.</dc:creator>
<dc:creator>Hirota, T.</dc:creator>
<dc:creator>Stanzione, M.</dc:creator>
<dc:creator>Ojarikre, O.</dc:creator>
<dc:creator>Maciulyte, V.</dc:creator>
<dc:creator>de Rooij, D.</dc:creator>
<dc:creator>Toth, A.</dc:creator>
<dc:creator>Keeney, S.</dc:creator>
<dc:creator>Turner, J. M.</dc:creator>
<dc:date>2017-09-15</dc:date>
<dc:identifier>doi:10.1101/172106</dc:identifier>
<dc:title><![CDATA[ATR is a multifunctional regulator of male mouse meiosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/190389v1?rss=1">
<title>
<![CDATA[
A Yeast Global Genetic Screen Reveals that Metformin Induces an Iron Deficiency-Like State 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/190389v1?rss=1"
</link>
<description><![CDATA[
We report here a simple and global strategy to map out gene functions and target pathways of drugs, toxins or other small molecules based on "homomer dynamics" Protein-fragment Complementation Assays (hdPCA). hdPCA measures changes in self-association (homomerization) of over 3,500 yeast proteins in yeast grown under different conditions. hdPCA complements genetic interaction measurements while eliminating confounding effects of gene ablation. We demonstrate that hdPCA accurately predicts the effects of two longevity and health-span-affecting drugs, immunosuppressant rapamycin and type II diabetes drug metformin, on cellular pathways. We also discovered an unsuspected global cellular response to metformin that resembles iron deficiency. This discovery opens a new avenue to investigate molecular mechanisms for the prevention or treatments of diabetes, cancers and other chronic diseases of aging.
]]></description>
<dc:creator>Stynen, B.</dc:creator>
<dc:creator>Abd-Rabbo, D.</dc:creator>
<dc:creator>Kowarzyk, J.</dc:creator>
<dc:creator>Miller-Fleming, L.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:creator>Michnick, S.</dc:creator>
<dc:date>2017-09-18</dc:date>
<dc:identifier>doi:10.1101/190389</dc:identifier>
<dc:title><![CDATA[A Yeast Global Genetic Screen Reveals that Metformin Induces an Iron Deficiency-Like State]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-09-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/191338v1?rss=1">
<title>
<![CDATA[
Non-canonical circadian oscillations in Drosophila S2 cells drive gene-expression cycles coupled to metabolic oscillations 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/191338v1?rss=1"
</link>
<description><![CDATA[
Circadian rhythms are cell-autonomous biological oscillations with a period of about 24 hours. Current models propose that transcriptional feedback loops are the principal mechanism for the generation of circadian oscillations. In these models, Drosophila S2 cells are generally regarded as  non-rhythmic cells, as they do not express several canonical circadian components. Using an unbiased multi-omics approach, we made the surprising discovery that Drosophila S2 cells do in fact display widespread daily rhythms. Transcriptomics and proteomics analyses revealed that hundreds of genes and their products are rhythmically expressed in a 24-hour cycle. Metabolomics analyses extended these findings and illustrated that central carbon metabolism and amino acid metabolism are the main pathways regulated in a rhythmic fashion. We thus demonstrate that daily genome-wide oscillations, coupled to metabolic cycles, take place in eukaryotic cells without the contribution of known circadian regulators.
]]></description>
<dc:creator>Rey, G.</dc:creator>
<dc:creator>Milev, N. B.</dc:creator>
<dc:creator>Valekunja, U. K.</dc:creator>
<dc:creator>Ch, R.</dc:creator>
<dc:creator>Ray, S.</dc:creator>
<dc:creator>Silva Dos Santos, M.</dc:creator>
<dc:creator>Nagy, A. D.</dc:creator>
<dc:creator>Antrobus, R.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Reddy, A. B.</dc:creator>
<dc:date>2017-09-20</dc:date>
<dc:identifier>doi:10.1101/191338</dc:identifier>
<dc:title><![CDATA[Non-canonical circadian oscillations in Drosophila S2 cells drive gene-expression cycles coupled to metabolic oscillations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-09-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/192757v1?rss=1">
<title>
<![CDATA[
Integrated analysis sheds light on evolutionary trajectories of young transcription start sites in the human genome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/192757v1?rss=1"
</link>
<description><![CDATA[
Previous studies revealed widespread transcription initiation and fast turnover of transcription start sites (TSSs) in mammalian genomes. Yet how new TSSs originate and how they evolve over time remain poorly understood. To address these questions, we analyzed [~]200,000 human TSSs by integrating evolutionary and functional genomic data, particularly focusing on TSSs that emerged in the primate lineages. We found that intrinsic factors of repetitive sequences and their proximity to established regulatory modules (extrinsic factors) contribute significantly to origin of new TSSs. In early periods, young TSSs experience rapid sequence evolution driven by endogenous mutational mechanisms that reduce the instability of associated repetitive sequences. In later periods, the regulatory functions of young TSSs are gradually modified, and with evolutionary changes subject to temporal (fewer regulatory changes in younger TSSs) and spatial constraints (fewer regulatory changes in more isolated TSSs). These findings advance our understanding of how regulatory innovations arise in the genome throughout evolution and highlight the roles of repetitive sequences in these processes.
]]></description>
<dc:creator>Li, C.</dc:creator>
<dc:creator>Lenhard, B.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:date>2017-09-22</dc:date>
<dc:identifier>doi:10.1101/192757</dc:identifier>
<dc:title><![CDATA[Integrated analysis sheds light on evolutionary trajectories of young transcription start sites in the human genome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/197699v1?rss=1">
<title>
<![CDATA[
Identifying developmentally important genes with single-cell RNA-seq from an embryo 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/197699v1?rss=1"
</link>
<description><![CDATA[
Single-cell RNA-seq has been established as a reliable and accessible technique enabling new types of analyses, such as identifying cell types and studying spatial and temporal gene expression variation and change at single-cell resolution. Recently, single-cell RNA-seq has been applied to developing embryos, which offers great potential for finding and characterising genes controlling the course of development along with their expression patterns. In this study, we applied single-cell RNA-seq to the 16-cell stage of the Ciona embryo, a marine chordate and performed a computational search for cell-specific gene expression patterns. We recovered many known expression patterns from our single-cell RNA-seq data and despite extensive previous screens, we succeeded in finding new cell-specific patterns, which we validated by in situ and single-cell qPCR.
]]></description>
<dc:creator>Ilsley, G. R.</dc:creator>
<dc:creator>Suyama, R.</dc:creator>
<dc:creator>Noda, T.</dc:creator>
<dc:creator>Satoh, N.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:date>2017-10-03</dc:date>
<dc:identifier>doi:10.1101/197699</dc:identifier>
<dc:title><![CDATA[Identifying developmentally important genes with single-cell RNA-seq from an embryo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/201004v1?rss=1">
<title>
<![CDATA[
Actin and Src-family kinases regulate nuclear YAP1 and its export 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/201004v1?rss=1"
</link>
<description><![CDATA[
The transcriptional regulator YAP1 is critical for the pathological activation of fibroblasts. In normal fibroblasts YAP1 is predominantly located in the cytoplasm, while in activated cancer-associated fibroblasts it exhibits nuclear localization and promotes the expression of many genes required for pro-tumorigenic functions. Here, we investigate the dynamics of YAP1 shuttling in normal and activated fibroblasts, using EYFP-YAP1, quantitative photo-bleaching methods, and mathematical modeling. We find that both 14-3-3 and TEAD binding modulate YAP1 shuttling, but neither affects nuclear import. Instead, we find that YAP1 serine phosphorylation is required for nuclear export. Furthermore, YAP1 nuclear accumulation in activated fibroblasts results from Src and actomyosin-dependent suppression of phosphorylated YAP1 export. Finally, we show that nuclear constrained YAP1, upon XPO1 depletion, remains sensitive to blockade of actomyosin function. Together, these data place nuclear export at the center of YAP1 regulation and indicate that the cytoskeleton can regulate YAP1 within the nucleus.
]]></description>
<dc:creator>Ege, N.</dc:creator>
<dc:creator>Dowbaj, A. M.</dc:creator>
<dc:creator>Jiang, M.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>Jenkins, R. P.</dc:creator>
<dc:creator>Sahai, E.</dc:creator>
<dc:date>2017-10-11</dc:date>
<dc:identifier>doi:10.1101/201004</dc:identifier>
<dc:title><![CDATA[Actin and Src-family kinases regulate nuclear YAP1 and its export]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/200154v1?rss=1">
<title>
<![CDATA[
WDR11-mediated Hedgehog signalling defects underlie a new ciliopathy related to Kallmann syndrome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/200154v1?rss=1"
</link>
<description><![CDATA[
WDR11 has been implicated in congenital hypogonadotropic hypogonadism (CHH) and Kallmann syndrome (KS), human developmental genetic disorders defined by delayed puberty and infertility. However, WDR11s role in development is poorly understood. Here we report that WDR11 modulates the Hedgehog (Hh) signalling pathway and is essential for ciliogenesis. Disruption of WDR11 expression in mouse and zebrafish results in phenotypic characteristics associated with defective Hh signalling, accompanied by dysgenesis of ciliated tissues. Wdr11 null mice also exhibit early onset obesity. We found that WDR11 shuttles from the cilium to the nucleus in response to Hh signalling. WDR11 was also observed to regulate the proteolytic processing of GLI3 and cooperate with EMX1 transcription factor to induce the expression of downstream Hh pathway genes and gonadotrophin releasing hormone production. The CHH/KS-associated human mutations result in loss-of-function of WDR11. Treatment with the Hh agonist purmorphamine partially rescued the WDR11-haploinsufficiency phenotypes. Our study reveals a novel class of ciliopathy caused by WDR11 mutations and suggests that CHH/KS may be a part of the human ciliopathy spectrum.
]]></description>
<dc:creator>Kim, Y.-J.</dc:creator>
<dc:creator>Osborn, D.</dc:creator>
<dc:creator>Lee, J.-Y.</dc:creator>
<dc:creator>Araki, M.</dc:creator>
<dc:creator>Araki, K.</dc:creator>
<dc:creator>Mohun, T.</dc:creator>
<dc:creator>Kansakoski, J. E. M.</dc:creator>
<dc:creator>Brandstack, N.</dc:creator>
<dc:creator>Kim, H.-T.</dc:creator>
<dc:creator>Miralles, F.</dc:creator>
<dc:creator>Kim, C.-H.</dc:creator>
<dc:creator>Brown, N. A.</dc:creator>
<dc:creator>Kim, H.-G.</dc:creator>
<dc:creator>Martinez-Barbera, J. P.</dc:creator>
<dc:creator>Ataliotis, P.</dc:creator>
<dc:creator>Raivio, T.</dc:creator>
<dc:creator>Layman, L. C.</dc:creator>
<dc:creator>Kim, S.-H.</dc:creator>
<dc:date>2017-10-10</dc:date>
<dc:identifier>doi:10.1101/200154</dc:identifier>
<dc:title><![CDATA[WDR11-mediated Hedgehog signalling defects underlie a new ciliopathy related to Kallmann syndrome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/202812v1?rss=1">
<title>
<![CDATA[
Plasmodium falciparum dipeptidyl aminopeptidase 3 activity is important for efficient erythrocyte invasion. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/202812v1?rss=1"
</link>
<description><![CDATA[
Parasite egress from infected erythrocytes and invasion of new erythrocytes are essential for the exponential asexual replication of the malaria parasite, and both processes are regulated and mediated by proteases. The putative cysteine protease dipeptidyl aminopeptidase 3 (DPAP3) was previously suggested to be essential for parasite egress, but little is known about its biological function. Here, we demonstrate that DPAP3 has proteolytic activity, but contrary to previously studied DPAPs, removal of its prodomain is not required for activation. Interestingly, P. falciparum DPAP3 localizes to merozoite apical organelles from which it is secreted immediately before egress. Using a conditional knock out approach coupled to complementation studies with wild type or mutant DPAP3, we show that DPAP3 activity is critical for efficient RBC invasion and overall parasite replication, and demonstrate that it does not play a role in parasite egress. Overall, this study establishes DPAP3 as a key regulator of erythrocyte invasion.
]]></description>
<dc:creator>Lehmann, C.</dc:creator>
<dc:creator>Tan, M. S. Y.</dc:creator>
<dc:creator>de Vries, L. E.</dc:creator>
<dc:creator>Russo, I.</dc:creator>
<dc:creator>Sanchez, M. I.</dc:creator>
<dc:creator>Goldberg, D. A.</dc:creator>
<dc:creator>Deu, E.</dc:creator>
<dc:date>2017-10-13</dc:date>
<dc:identifier>doi:10.1101/202812</dc:identifier>
<dc:title><![CDATA[Plasmodium falciparum dipeptidyl aminopeptidase 3 activity is important for efficient erythrocyte invasion.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/208124v1?rss=1">
<title>
<![CDATA[
Data Science Issues in Understanding Protein-RNA Interactions 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/208124v1?rss=1"
</link>
<description><![CDATA[
An interplay of experimental and computational methods is required to achieve a comprehensive understanding of protein-RNA interactions. Crosslinking and immunoprecipitation (CLIP) identifies endogenous interactions by sequencing RNA fragments that co-purify with a selected RBP under stringent conditions. Here we focus on approaches for the analysis of resulting data and appraise the methods for peak calling, visualisation, analysis and computational modelling of protein-RNA binding sites. We advocate a combined assessment of cDNA complexity and specificity for data quality control. Moreover, we demonstrate the value of analysing sequence motif enrichment in peaks assigned from CLIP data, and of visualising RNA maps, which examine the positional distribution of peaks around regulated landmarks in transcripts. We use these to assess how variations in CLIP data quality, and in different peak calling methods, affect the insights into regulatory mechanisms. We conclude by discussing future opportunities for the computational analysis of protein-RNA interaction experiments.
]]></description>
<dc:creator>Chakrabarti, A. M.</dc:creator>
<dc:creator>Haberman, N.</dc:creator>
<dc:creator>Praznik, A.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:date>2017-10-25</dc:date>
<dc:identifier>doi:10.1101/208124</dc:identifier>
<dc:title><![CDATA[Data Science Issues in Understanding Protein-RNA Interactions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/212175v1?rss=1">
<title>
<![CDATA[
Detection of cytosolic Shigella flexneri via a C-terminal triple-arginine motif of GBP1 inhibits actin-based motility 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/212175v1?rss=1"
</link>
<description><![CDATA[
Dynamin-like guanylate binding proteins (GBPs) are gamma interferon (IFN{gamma})-inducible host defense proteins that can associate with cytosol-invading bacterial pathogens. Mouse GBPs promote the lytic destruction of targeted bacteria in the host cell cytosol but the antimicrobial function of human GBPs and the mechanism by which these proteins associate with cytosolic bacteria are poorly understood. Here, we demonstrate that human GBP1 is unique amongst the seven human GBP paralogs in its ability to associate with at least two cytosolic Gram-negative bacteria, Burkholderia thailandensis and Shigella flexneri. Rough lipopolysaccharide (LPS) mutants of S. flexneri co-localize with GBP1 less frequently than wildtype S. flexneri, suggesting that host recognition of O-antigen promotes GBP1 targeting to Gram-negative bacteria. The targeting of GBP1 to cytosolic bacteria, via a unique triple-arginine motif present in its C-terminus, promotes the co-recruitment of four additional GBP paralogs (GBP2, GBP3, GBP4 and GBP6). GBP1-decorated Shigella replicate but fail to form actin tails leading to their intracellular aggregation. Consequentially, wildtype but not the triple-arginine GBP1 mutant restricts S. flexneri cell-to-cell spread. Furthermore, human-adapted S. flexneri, through the action of one its secreted effectors, IpaH9.8, is more resistant to GBP1 targeting than the non-human-adapted bacillus B. thailandensis. These studies reveal that human GBP1 uniquely functions as an intracellular  glue trap inhibiting the cytosolic movement of normally actin-propelled Gram-negative bacteria. In response to this powerful human defense program S. flexneri has evolved an effective counter-defense to restrict GBP1 recruitment.nnImportanceSeveral pathogenic bacterial species evolved to invade, reside and replicate inside the cytosol of their host cells. One adaptation common to most cytosolic bacterial pathogens is the ability to co-opt the hosts actin polymerization machinery, in order to generate force for intracellular movement. This actin-based motility enables Gram-negative bacteria such as Shigella to propel themselves into neighboring cells thereby spreading from host cell to host cell without exiting the intracellular environment. Here, we show that the human protein GBP1 acts as a cytosolic  glue trap capturing cytosolic Gram-negative bacteria through a unique protein motif and preventing disseminated infections in cell culture models. To escape from this GBP1-mediated host defense, Shigella employs a virulence factor that prevents or dislodges the association of GBP1 with cytosolic bacteria. Thus, therapeutic strategies to restore GBP1 binding to Shigella may lead to novel treatment options for shigellosis in the future.
]]></description>
<dc:creator>Piro, A.</dc:creator>
<dc:creator>Hernandez, D.</dc:creator>
<dc:creator>Luoma, S.</dc:creator>
<dc:creator>Feeley, E.</dc:creator>
<dc:creator>Finethy, R.</dc:creator>
<dc:creator>Yirga, A.</dc:creator>
<dc:creator>Frickel, E.</dc:creator>
<dc:creator>Lesser, C.</dc:creator>
<dc:creator>Coers, J.</dc:creator>
<dc:date>2017-11-01</dc:date>
<dc:identifier>doi:10.1101/212175</dc:identifier>
<dc:title><![CDATA[Detection of cytosolic Shigella flexneri via a C-terminal triple-arginine motif of GBP1 inhibits actin-based motility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/213827v1?rss=1">
<title>
<![CDATA[
Coherent olfactory bulb gamma oscillations arise from coupling independent columnar oscillators 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/213827v1?rss=1"
</link>
<description><![CDATA[
Spike timing-based representations of sensory information depend on embedded dynamical frameworks within neuronal networks that establish the rules of local computation and interareal communication. Here, we investigated the dynamical properties of olfactory bulb circuitry in mice of both sexes using microelectrode array recordings from slice and in vivo preparations. Neurochemical activation or optogenetic stimulation of sensory afferents evoked persistent gamma oscillations in the local field potential. These oscillations arose from slower, GABA(A) receptor-independent intracolumnar oscillators coupled by GABA(A)-ergic synapses into a faster, broadly coherent network oscillation. Consistent with the theoretical properties of coupled-oscillator networks, the spatial extent of zero-phase coherence was bounded in slices by the reduced density of lateral interactions. The intact in vivo network, however, exhibited long-range lateral interactions that suffice in simulation to enable zero-phase gamma coherence across the olfactory bulb. The timing of action potentials in a subset of principal neurons was phase-constrained with respect to evoked gamma oscillations. Coupled-oscillator dynamics in olfactory bulb thereby enable a common clock, robust to biological heterogeneities, that is capable of supporting gamma-band spike synchronization and phase coding across the ensemble of activated principal neurons.

New & NoteworthyOdor stimulation evokes rhythmic gamma oscillations in the field potential of the olfactory bulb, but the dynamical mechanisms governing these oscillations have remained unclear. Establishing these mechanisms is important, as they determine the biophysical capacities of the bulbar circuit to, for example, maintain zero-phase coherence across a spatially extended network, or coordinate the timing of action potentials in principal neurons. These properties in turn constrain and suggest hypotheses of sensory coding.
]]></description>
<dc:creator>Peace, S. T.</dc:creator>
<dc:creator>Johnson, B. C.</dc:creator>
<dc:creator>Li, G.</dc:creator>
<dc:creator>Kaiser, M. E.</dc:creator>
<dc:creator>Fukunaga, I.</dc:creator>
<dc:creator>Schaefer, A. T.</dc:creator>
<dc:creator>Molnar, A. C.</dc:creator>
<dc:creator>Cleland, T. A.</dc:creator>
<dc:date>2017-11-03</dc:date>
<dc:identifier>doi:10.1101/213827</dc:identifier>
<dc:title><![CDATA[Coherent olfactory bulb gamma oscillations arise from coupling independent columnar oscillators]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/216879v1?rss=1">
<title>
<![CDATA[
A modular transcriptional signature identifies phenotypic heterogeneity of human tuberculosis infection 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/216879v1?rss=1"
</link>
<description><![CDATA[
Whole blood transcriptional signatures distinguishing active tuberculosis patients from asymptomatic latently infected individuals exist. Consensus has not been achieved regarding the optimal reduced gene sets as diagnostic biomarkers that also achieve discrimination from other diseases. Here we show a blood transcriptional signature of active tuberculosis using RNA-Seq, confirming microarray results, that discriminates active tuberculosis from latently infected and healthy individuals, validating this signature in an independent cohort. Using an advanced modular approach, we utilise information from the entire transcriptome, which includes over-abundance of type I interferon-inducible genes and under-abundance of IFNG and TBX21, to develop a signature that discriminates active tuberculosis patients from latently infected individuals, or those with acute viral and bacterial infections. We suggest methods targeting gene selection across multiple discriminant modules can improve development of diagnostic biomarkers with improved performance. Finally, utilising the modular approach we demonstrate dynamic heterogeneity in a longitudinal study of recent tuberculosis contacts.
]]></description>
<dc:creator>Singhania, A.</dc:creator>
<dc:creator>Verma, R.</dc:creator>
<dc:creator>Graham, C. M.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Trang, T.</dc:creator>
<dc:creator>Richarsdon, M.</dc:creator>
<dc:creator>Lecine, P.</dc:creator>
<dc:creator>Leissner, P.</dc:creator>
<dc:creator>Berry, M. P. R.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Kaiser, K.</dc:creator>
<dc:creator>Rodrigue, M.</dc:creator>
<dc:creator>Woltmann, G.</dc:creator>
<dc:creator>Haldar, P.</dc:creator>
<dc:creator>O'Garra, A.</dc:creator>
<dc:date>2017-11-09</dc:date>
<dc:identifier>doi:10.1101/216879</dc:identifier>
<dc:title><![CDATA[A modular transcriptional signature identifies phenotypic heterogeneity of human tuberculosis infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/222588v1?rss=1">
<title>
<![CDATA[
Active sampling state dynamically enhances olfactory bulb odor representation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/222588v1?rss=1"
</link>
<description><![CDATA[
The olfactory bulb (OB) is the very first site of odor information processing, yet a wealth of contextual and learned information has been described in its activity. To investigate the mechanistic basis of contextual modulation, we use whole-cell recordings to measure odor responses across rapid (<30 min) learning episodes in identified mitral/tufted cells (MTCs). Across these learning episodes, we found that diverse response changes occur already during the first sniff cycle. Motivated mice develop active sniffing strategies across learning, and it is this change of active sampling state that dynamically modulates odor responses, resulting in enhanced discriminability and detectability of odor representation with learning. Evoking fast sniffing in different behavioral states demonstrates that response changes during active sampling exceed those predicted from purely feed-forward input. Finally, response changes are highly correlated in tufted cells, but not mitral cells, indicating cell-type specificity in the effect of active sampling, and resulting in increased odor detectability in the tufted and enhanced discriminability in the mitral cell population over the rapid learning episodes. Altogether, we show that active sampling state is a crucial component in modulating and enhancing olfactory bulb responsiveness on rapid timescales.
]]></description>
<dc:creator>Jordan, R.</dc:creator>
<dc:creator>Fukunaga, I.</dc:creator>
<dc:creator>Kollo, M.</dc:creator>
<dc:creator>Schaefer, A. T.</dc:creator>
<dc:date>2017-11-21</dc:date>
<dc:identifier>doi:10.1101/222588</dc:identifier>
<dc:title><![CDATA[Active sampling state dynamically enhances olfactory bulb odor representation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/223776v1?rss=1">
<title>
<![CDATA[
BCL11A interacts with SOX2 to control the expression of epigenetic regulators in lung squamous cell carcinoma 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/223776v1?rss=1"
</link>
<description><![CDATA[
Patients diagnosed with lung squamous cell carcinoma (LUSC) have limited targeted therapeutic options. We report here the identification and characterisation of the transcriptional regulator, BCL11A, as a LUSC oncogene. Analysis of cancer genomics datasets revealed BCL11A to be upregulated in LUSC but not lung adenocarcinoma (LUAD). We demonstrated that knockdown of BCL11A in LUSC cell lines abolished xenograft tumour growth and its overexpression in vivo led to lung airway hyperplasia and the development of reserve cell hyperplastic lesions. In addition, deletion of Bcl11a in the tracheal basal cells abolished the development of tracheosphere organoids while its overexpression led to solid tracheospheres with a squamous phenotype. At the molecular level we found BCL11A to be a target of SOX2 and we show that it is required for the oncogenic role of SOX2 in LUSC. Furthermore, we showed that BCL11A and SOX2 interact at the protein level and that together they co-regulated the expression of several transcription factors. We demonstrate that pharmacological inhibition of SETD8, a gene co-regulated by BCL11A and SOX2, alone or in combination with cisplatin treatment, shows significant selectivity to LUSC in comparison to LUAD cells. Collectively, these results indicate that the disruption of the BCL11A-SOX2 transcriptional program provides a future framework for the development of targeted therapeutic intervention for LUSC patients.
]]></description>
<dc:creator>Lazarus, K. A.</dc:creator>
<dc:creator>Hadi, F.</dc:creator>
<dc:creator>Zambon, E.</dc:creator>
<dc:creator>Bach, K.</dc:creator>
<dc:creator>Santolla, M.-F.</dc:creator>
<dc:creator>Watson, J. K.</dc:creator>
<dc:creator>Correia, L. L.</dc:creator>
<dc:creator>Das, M.</dc:creator>
<dc:creator>Ugur, R.</dc:creator>
<dc:creator>Pensa, S.</dc:creator>
<dc:creator>Becker, L.</dc:creator>
<dc:creator>Campos, L. S.</dc:creator>
<dc:creator>Ladds, G.</dc:creator>
<dc:creator>Liu, P.</dc:creator>
<dc:creator>Evan, G.</dc:creator>
<dc:creator>McCaughan, F.</dc:creator>
<dc:creator>Le Quesne, J.</dc:creator>
<dc:creator>Lee, J.-H.</dc:creator>
<dc:creator>Calado, D.</dc:creator>
<dc:creator>Khaled, W. T.</dc:creator>
<dc:date>2017-11-22</dc:date>
<dc:identifier>doi:10.1101/223776</dc:identifier>
<dc:title><![CDATA[BCL11A interacts with SOX2 to control the expression of epigenetic regulators in lung squamous cell carcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/223875v1?rss=1">
<title>
<![CDATA[
A hidden Markov model approach to characterizing the photo-switching behavior of fluorophores 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/223875v1?rss=1"
</link>
<description><![CDATA[
Fluorescing molecules (fluorophores) that stochastically switch between photon-emitting and dark states underpin some of the most celebrated advancements in super-resolution microscopy. While this stochastic behavior has been heavily exploited, full characterization of the underlying models can potentially drive forward further imaging methodologies. Under the assumption that fluorophores move between fluorescing and dark states as continuous time Markov processes, the goal is to use a sequence of images to select a model and estimate the transition rates. We use a hidden Markov model to relate the observed discrete time signal to the hidden continuous time process. With imaging involving several repeat exposures of the fluorophore, we show the observed signal depends on both the current and past states of the hidden process, producing emission probabilities that depend on the transition rate parameters to be estimated. To tackle this unusual coupling of the transition and emission probabilities, we conceive transmission (transition-emission) matrices that capture all dependencies of the model. We provide a scheme of computing these matrices and adapt the forward-backward algorithm to compute a likelihood which is readily optimized to provide rate estimates. When confronted with several model proposals, combining this procedure with the Bayesian Information Criterion provides accurate model selection.
]]></description>
<dc:creator>Patel, L.</dc:creator>
<dc:creator>Gustafsson, N.</dc:creator>
<dc:creator>Lin, Y.</dc:creator>
<dc:creator>Ober, R.</dc:creator>
<dc:creator>Henriques, R.</dc:creator>
<dc:creator>Cohen, E.</dc:creator>
<dc:date>2017-11-22</dc:date>
<dc:identifier>doi:10.1101/223875</dc:identifier>
<dc:title><![CDATA[A hidden Markov model approach to characterizing the photo-switching behavior of fluorophores]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/224550v1?rss=1">
<title>
<![CDATA[
A single cycle influenza virus coated in H7 hemagglutinin provides heterotypic protection and neutralising antibody responses to both glycoproteins. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/224550v1?rss=1"
</link>
<description><![CDATA[
A non-replicating form of pseudotyped influenza virus, inactivated by suppression of the hemagglutinin signal sequence (S-FLU), can act as a broadly protective vaccine. S-FLU can infect for a single round only, and induces heterotypic protection predominantly through activation of cross-reactive T cells in the lung. Unlike the licensed live attenuated virus, it cannot reassort a pandemic HA into seasonal influenza. Here we present data on four new forms of S-FLU coated with the H7 hemagglutinins from A/Anhui/1/2013 and A/Shanghai/1/2013, H7N9 viruses that emerged recently in China, and A/Netherlands/219/2003 and A/New York/107/2003. We show that vaccination in the lung induced a strong local CD8 T cell response and protected against heterosubtypic X31 (H3N2) virus and highly virulent PR8 (H1N1), but not influenza B virus. Lung vaccination also induced a strong neutralising antibody response to the encoded neuraminidase. If given at higher dose in the periphery, H7 S-FLU induced a specific neutralising antibody response to H7 hemagglutinin coating the particle. Polyvalent vaccination with mixed H7 S-FLU induced a broadly neutralising antibody response to all four H7 strains. S-FLU is a versatile vaccine candidate that could be rapidly mobilized ahead of a new pandemic threat.
]]></description>
<dc:creator>Powell, T. J.</dc:creator>
<dc:creator>Rijal, P.</dc:creator>
<dc:creator>McEwen-Smith, R. M.</dc:creator>
<dc:creator>Byun, H.</dc:creator>
<dc:creator>Hardwick, M.</dc:creator>
<dc:creator>Schimanski, L. M.</dc:creator>
<dc:creator>Huang, K.-Y. A.</dc:creator>
<dc:creator>Daniels, R. S.</dc:creator>
<dc:creator>Townsend, A. R. M.</dc:creator>
<dc:date>2017-11-27</dc:date>
<dc:identifier>doi:10.1101/224550</dc:identifier>
<dc:title><![CDATA[A single cycle influenza virus coated in H7 hemagglutinin provides heterotypic protection and neutralising antibody responses to both glycoproteins.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/164905v1?rss=1">
<title>
<![CDATA[
CYRI (FAM49) proteins are local inhibitors of Scar/WAVE induced lamellipodia that bind directly to active Rac1 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/164905v1?rss=1"
</link>
<description><![CDATA[
Actin-based protrusions driving cell migration are reinforced through positive feedback, but it is unclear how the cell restricts the eventual size of a protrusion or limits positive signals to cause splitting or retraction. We have identified an evolutionarily conserved regulator of the protrusion machinery, which we name CYRI (CYFIP-related Rac interacting) protein. CYRI shows sequence similarity to the Scar/WAVE complex member CYFIP in a Domain of Unknown Function, DUF1394. CYRI binds specifically to activated Rac1 via a common motif shared with CYFIP, establishing DUF1394 as a new Rac1 binding domain. CYRI-depleted cells have broad, Scar/WAVE-enriched lamellipodia and enhanced Rac1 signaling. Conversely, CYRI overexpression suppresses spreading and dramatically sharpens protrusions into unproductive needles. CYRI proteins use dynamic inhibition of Scar/WAVE induced actin to focus positive protrusion signals and regulate pseudopod complexity. CYRI behaves like a "local inhibitor" predicted and described in widely accepted mathematical models, but not previously identified in living cells.
]]></description>
<dc:creator>Fort, L.</dc:creator>
<dc:creator>Batista, J.</dc:creator>
<dc:creator>Thomason, P. A.</dc:creator>
<dc:creator>Spence, H. J.</dc:creator>
<dc:creator>Greaves, J.</dc:creator>
<dc:creator>Martin, K. J.</dc:creator>
<dc:creator>Anderson, K. I.</dc:creator>
<dc:creator>Brown, P.</dc:creator>
<dc:creator>Lilla, S.</dc:creator>
<dc:creator>Neilson, M. P.</dc:creator>
<dc:creator>Tafelmeyer, P.</dc:creator>
<dc:creator>Zanivan, S.</dc:creator>
<dc:creator>Ismail, S.</dc:creator>
<dc:creator>Tomkinson, N. C. O.</dc:creator>
<dc:creator>Chamberlain, L. H.</dc:creator>
<dc:creator>Insall, R. H.</dc:creator>
<dc:creator>Machesky, L. M.</dc:creator>
<dc:date>2017-11-29</dc:date>
<dc:identifier>doi:10.1101/164905</dc:identifier>
<dc:title><![CDATA[CYRI (FAM49) proteins are local inhibitors of Scar/WAVE induced lamellipodia that bind directly to active Rac1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/226969v1?rss=1">
<title>
<![CDATA[
Sniff-invariant intensity perception using olfactory bulb coding of inhalation dynamics 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/226969v1?rss=1"
</link>
<description><![CDATA[
For stable perception of odor intensity, there must exist a neural correlate that is invariant across other parameters, such as the highly variable sniff cycle. Previous hypotheses suggest that variance in inhalation dynamics alters odor concentration profiles in the naris despite a constant environmental concentration. Using whole cell recordings in the olfactory bulb of awake mice, we directly demonstrate that increased sniffing frequency mimics the effect of odor concentration increase at the level of both mitral and tufted cell (MTC) firing rate responses and temporal responses. In contrast, we find that mice are capable of discriminating concentrations within short timescales despite highly variable sniffing behavior. We reason that the only way mice can differentiate a change in sniff parameters and a change in concentration is to encode information about the inhalation parameters in parallel. While conceivably this could be achieved via corollary discharge from respiration control centres, we find that the sniff-driven activity of MTCs without odor input is informative of the kind of inhalation that just occurred, allowing rapid detection of a change in inhalation. Thus, a possible reason for sniff modulation of the early olfactory system may be to inform downstream centres of nasal flow dynamics, so that an inference can be made about environmental concentration independent of sniff variance.
]]></description>
<dc:creator>Jordan, R.</dc:creator>
<dc:creator>Kollo, M.</dc:creator>
<dc:creator>Schaefer, A. T.</dc:creator>
<dc:date>2017-11-29</dc:date>
<dc:identifier>doi:10.1101/226969</dc:identifier>
<dc:title><![CDATA[Sniff-invariant intensity perception using olfactory bulb coding of inhalation dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/229203v1?rss=1">
<title>
<![CDATA[
Regionalization of the nervous system requires axial allocation prior to neural lineage commitment 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/229203v1?rss=1"
</link>
<description><![CDATA[
SummaryNeural induction in vertebrates generates a central nervous system that extends the rostral-caudal length of the body. The prevailing view is that neural cells are initially induced with anterior (forebrain) identity, with caudalising signals then converting a proportion to posterior fates (spinal cord). To test this model, we used chromatin accessibility assays to define how cells adopt region-specific neural fates. Together with genetic and biochemical perturbations this identified a developmental time window in which genome-wide chromatin remodeling events preconfigure epiblast cells for neural induction. Contrary to the established model, this revealed that cells commit to a regional identity before acquiring neural identity. This "primary regionalization" allocates cells to anterior or posterior regions of the nervous system, explaining how cranial and spinal neurons are generated at appropriate axial positions. These findings prompt a revision to models of neural induction and support the proposed dual evolutionary origin of the vertebrate central nervous system.
]]></description>
<dc:creator>Metzis, V.</dc:creator>
<dc:creator>Steinhauser, S.</dc:creator>
<dc:creator>Pakanavicius, E.</dc:creator>
<dc:creator>Gouti, M.</dc:creator>
<dc:creator>Stamataki, D.</dc:creator>
<dc:creator>Lovell-Badge, R.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:date>2017-12-04</dc:date>
<dc:identifier>doi:10.1101/229203</dc:identifier>
<dc:title><![CDATA[Regionalization of the nervous system requires axial allocation prior to neural lineage commitment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-12-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/235101v1?rss=1">
<title>
<![CDATA[
AP-4 mediated ATG9A sorting underlies axonal and autophagosome biogenesis defects in a mouse model of AP-4 deficiency syndrome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/235101v1?rss=1"
</link>
<description><![CDATA[
Adaptor protein (AP) complexes have critical roles in transmembrane protein sorting. AP-4 remains poorly understood in the brain despite its loss of function leading to a hereditary spastic paraplegia termed AP-4 deficiency syndrome. Here we demonstrate that knockout (KO) of AP-4 in a mouse model leads to thinning of the corpus callosum and ventricular enlargement, anatomical defects previously described in patients. At the cellular level, we find that AP-4 KO leads to defects in axonal extension and branching, in addition to aberrant distal swellings. Interestingly, we show that ATG9A, a key protein in autophagosome maturation, is critically dependent on AP-4 for its sorting from the trans-golgi network. Failure of AP-4 mediated ATG9A sorting results in its dramatic retention in the trans-golgi network in vitro and in vivo leading to a specific reduction of the axonal pool of ATG9A. As a result, autophagosome biogenesis is aberrant in the axon of AP-4 deficient neurons. The specific alteration to axonal integrity and axonal autophagosome maturation in AP-4 knockout neurons may underpin the pathology of AP-4 deficiency.
]]></description>
<dc:creator>Ivankovic, D.</dc:creator>
<dc:creator>Lopez-Domenech, G.</dc:creator>
<dc:creator>Drew, J.</dc:creator>
<dc:creator>Tooze, S. A.</dc:creator>
<dc:creator>Kittler, J. T.</dc:creator>
<dc:date>2017-12-16</dc:date>
<dc:identifier>doi:10.1101/235101</dc:identifier>
<dc:title><![CDATA[AP-4 mediated ATG9A sorting underlies axonal and autophagosome biogenesis defects in a mouse model of AP-4 deficiency syndrome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-12-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/237602v1?rss=1">
<title>
<![CDATA[
PolNet Analysis: a software tool for the quantification of network-level endothelial cell polarity and blood flow during vascular remodelling 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/237602v1?rss=1"
</link>
<description><![CDATA[
In this paper, we present PolNet, an open source software tool for the study of blood flow and cell-level biological activity during vessel morphogenesis. We provide an image acquisition, segmentation, and analysis protocol to quantify endothelial cell polarity in entire in vivo vascular networks. In combination, we use computational fluid dynamics to characterise the haemodynamics of the vascular networks under study. The tool enables, for the first time, network-level analysis of polarity and flow for individual endothelial cells. To date, PolNet has proven invaluable for the study of endothelial cell polarisation and migration during vascular patterning, as demonstrated by our recent papers [1, 2]. Additionally, the tool can be easily extended to correlate blood flow with other experimental observations at the cellular/molecular level. We release the source code of our tool under the LGPL licence.
]]></description>
<dc:creator>Bernabeu, M. O.</dc:creator>
<dc:creator>Jones, M. L.</dc:creator>
<dc:creator>Nash, R. W.</dc:creator>
<dc:creator>Pezzarossa, A.</dc:creator>
<dc:creator>Coveney, P. V.</dc:creator>
<dc:creator>Gerhardt, H.</dc:creator>
<dc:creator>Franco, C. A.</dc:creator>
<dc:date>2017-12-22</dc:date>
<dc:identifier>doi:10.1101/237602</dc:identifier>
<dc:title><![CDATA[PolNet Analysis: a software tool for the quantification of network-level endothelial cell polarity and blood flow during vascular remodelling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/242974v1?rss=1">
<title>
<![CDATA[
The Homeostatic Logic of Reward 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/242974v1?rss=1"
</link>
<description><![CDATA[
Energy homeostasis depends on behavior to predictively regulate metabolic states within narrow bounds. Here we review three theories of homeostatic control and ask how they provide insight into the circuitry underlying energy homeostasis. We offer two contributions. First, we detail how control theory and reinforcement learning are applied to homeostatic control. We show how these schemes rest on implausible assumptions; either via circular definitions, unprincipled drive functions, or by ignoring environmental volatility. We argue active inference can elude these shortcomings while retaining important features of each model. Second, we review the neural basis of energetic control. We focus on a subset of arcuate subpopulations that project directly to, and are thus in a privileged position to opponently modulate, dopaminergic cells as a function of energetic predictions over a spectrum of time horizons. We discuss how this can be interpreted under these theories, and how this can resolve paradoxes that have arisen. We propose this circuit constitutes a homeostatic-reward interface that underwrites the conjoint optimisation of physiological and behavioural homeostasis.
]]></description>
<dc:creator>Morville, T.</dc:creator>
<dc:creator>Friston, K.</dc:creator>
<dc:creator>Burdakov, D.</dc:creator>
<dc:creator>Siebner, H. R.</dc:creator>
<dc:creator>Hulme, O. J.</dc:creator>
<dc:date>2018-01-05</dc:date>
<dc:identifier>doi:10.1101/242974</dc:identifier>
<dc:title><![CDATA[The Homeostatic Logic of Reward]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/246124v1?rss=1">
<title>
<![CDATA[
Characterization of P. falciparum dipeptidyl aminopeptidase 3 specificity identifies different amino acid preferences between peptide-based substrates and inhibitors. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/246124v1?rss=1"
</link>
<description><![CDATA[
Malarial dipeptidyl aminopeptidases (DPAPs) are cysteine proteases important for parasite development thus making them attractive drug targets. In order to develop inhibitors specific to the parasite enzymes it is necessary to map the determinants of substrate specificity of the parasite enzymes and its mammalian homologue cathepsin C (CatC). Here, we screened peptide-based libraries of substrates and covalent inhibitors to characterize the differences in specificity between parasite DPAPs and CatC, and used this information to develop highly selective DPAP1 and DPAP3 inhibitors. Interestingly, while the primary amino acid specificity of a protease is often used to develop potent inhibitors, we show that equally potent and highly specific inhibitors can be developed based on the sequences of non-optimal peptide substrates. Importantly, analysis of previously published data about the specificity of other proteases also unveiled significant discrepancies in the amino acid preference between substrates and inhibitors. In this article, we also discuss important structural and theoretical reasons that might account for these discrepancies. Overall, this study illustrates that focusing the development of protease inhibitors solely on substrate specificity might overlook important structural features that can be exploited to develop highly potent and selective compounds.
]]></description>
<dc:creator>Deu, E.</dc:creator>
<dc:creator>de Vries, L. E.</dc:creator>
<dc:creator>Sanchez, M. I.</dc:creator>
<dc:creator>Groborz, K.</dc:creator>
<dc:creator>Kuppens, L.</dc:creator>
<dc:creator>Poreba, M.</dc:creator>
<dc:creator>Lehmann, C.</dc:creator>
<dc:creator>Yuan, F.</dc:creator>
<dc:creator>Arastu-Kapur, S.</dc:creator>
<dc:creator>Bogyo, M.</dc:creator>
<dc:creator>Drag, M.</dc:creator>
<dc:date>2018-01-11</dc:date>
<dc:identifier>doi:10.1101/246124</dc:identifier>
<dc:title><![CDATA[Characterization of P. falciparum dipeptidyl aminopeptidase 3 specificity identifies different amino acid preferences between peptide-based substrates and inhibitors.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/254623v1?rss=1">
<title>
<![CDATA[
A role for a conserved kinase in the transcriptional control of methionine biosynthesis in Escherichia coli experiencing sustained nitrogen starvation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/254623v1?rss=1"
</link>
<description><![CDATA[
The initial adaptive transcriptional response to nitrogen (N) starvation in Escherichia coli involves large-scale alterations to the transcriptome mediated by the transcription activator, NtrC. One of the NtrC-activated genes is yeaG, which encodes a conserved bacterial kinase. Although it is known that YeaG is required for optimal survival under sustained N starvation, the molecular basis by which YeaG benefits N starved E. coli remains elusive. By combining transcriptomics with targeted metabolomics analyses, we demonstrate that the methionine biosynthesis pathway becomes transcriptionally dysregulated in{Delta} yeaG bacteria experiencing sustained N starvation. This results in the aberrant and energetically costly biosynthesis of methionine and associated metabolites under sustained N starvation with detrimental consequences to cell viability. It appears the activity of the master transcriptional repressor of methionine biosynthesis genes, MetJ, is compromised in{Delta} yeaG bacteria under sustained N starvation, resulting in transcriptional derepression of MetJ-regulated genes. The results suggest that YeaG is a novel regulatory factor and functions as a molecular brake in the transcriptional control of both the NtrC-regulon and methionine biosynthesis genes in E. coli experiencing sustained N starvation.
]]></description>
<dc:creator>Switzer, A.</dc:creator>
<dc:creator>Evangelopoulos, D.</dc:creator>
<dc:creator>Figueira, R.</dc:creator>
<dc:creator>de Carvalho, L.</dc:creator>
<dc:creator>Brown, D.</dc:creator>
<dc:creator>Wigneshweraraj, S.</dc:creator>
<dc:date>2018-01-27</dc:date>
<dc:identifier>doi:10.1101/254623</dc:identifier>
<dc:title><![CDATA[A role for a conserved kinase in the transcriptional control of methionine biosynthesis in Escherichia coli experiencing sustained nitrogen starvation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/256420v1?rss=1">
<title>
<![CDATA[
Predicting Human Protein Function with Multi-task Deep Neural Networks 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/256420v1?rss=1"
</link>
<description><![CDATA[
Machine learning methods for protein function prediction are urgently needed, especially now that a substantial fraction of known sequences remains unannotated despite the extensive use of functional assignments based on sequence similarity. One major bottleneck supervised learning faces in protein function prediction is the structured, multi-label nature of the problem, because biological roles are represented by lists of terms from hierarchically organised controlled vocabularies such as the Gene Ontology. In this work, we build on recent developments in the area of deep learning and investigate the usefulness of multi-task deep neural networks (MTDNN), which consist of upstream shared layers upon which are stacked in parallel as many independent modules (additional hidden layers with their own output units) as the number of output GO terms (the tasks). MTDNN learns individual tasks partially using shared representations and partially from task-specific characteristics. When no close homologues with experimentally validated functions can be identified, MTDNN gives more accurate predictions than baseline methods based on annotation frequencies in public databases or homology transfers. More importantly, the results show that MTDNN binary classification accuracy is higher than alternative machine learning-based methods that do not exploit commonalities and differences among prediction tasks. Interestingly, compared with a single-task predictor, the performance improvement is not linearly correlated with the number of tasks in MTDNN, but medium size models provide more improvement in our case. One of advantages of MTDNN is that given a set of features, there is no requirement for MTDNN to have a bootstrap feature selection procedure as what traditional machine learning algorithms do. Overall, the results indicate that the proposed MTDNN algorithm improves the performance of protein function prediction. On the other hand, there is still large room for deep learning techniques to further enhance prediction ability.
]]></description>
<dc:creator>Fa, R.</dc:creator>
<dc:creator>Cozzetto, D.</dc:creator>
<dc:creator>Wan, C.</dc:creator>
<dc:creator>Jones, D. T.</dc:creator>
<dc:date>2018-01-30</dc:date>
<dc:identifier>doi:10.1101/256420</dc:identifier>
<dc:title><![CDATA[Predicting Human Protein Function with Multi-task Deep Neural Networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/256008v1?rss=1">
<title>
<![CDATA[
Machine learning models in electronic health records can outperform conventional survival models for predicting patient mortality in coronary artery disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/256008v1?rss=1"
</link>
<description><![CDATA[
Prognostic modelling is important in clinical practice and epidemiology for patient management and research. Electronic health records (EHR) provide large quantities of data for such models, but conventional epidemiological approaches require significant researcher time to implement. Expert selection of variables, fine-tuning of variable transformations and interactions, and imputing missing values in datasets are time-consuming and could bias subsequent analysis, particularly given that missingness in EHR is both high, and may carry meaning.nnUsing a cohort of over 80,000 patients from the CALIBER programme, we performed a systematic comparison of several machine-learning approaches in EHR. We used Cox models and random survival forests with and without imputation on 27 expert-selected variables to predict all-cause mortality. We also used Cox models, random forests and elastic net regression on an extended dataset with 586 variables to build prognostic models and identify novel prognostic factors without prior expert input.nnWe observed that data-driven models used on an extended dataset can outperform conventional models for prognosis, without data preprocessing or imputing missing values, and with no need to scale or transform continuous data. An elastic net Cox regression based with 586 unimputed variables with continuous values discretised achieved a C-index of 0.801 (bootstrapped 95% CI 0.799 to 0.802), compared to 0.793 (0.791 to 0.794) for a traditional Cox model comprising 27 expert-selected variables with imputation for missing values.nnWe also found that data-driven models allow identification of novel prognostic variables; that the absence of values for particular variables carries meaning, and can have significant implications for prognosis; and that variables often have a nonlinear association with mortality, which discretised Cox models and random forests can elucidate.nnThis demonstrates that machine-learning approaches applied to raw EHR data can be used to build reliable models for use in research and clinical practice, and identify novel predictive variables and their effects to inform future research.
]]></description>
<dc:creator>Steele, A. J.</dc:creator>
<dc:creator>Cakiroglu, S. A.</dc:creator>
<dc:creator>Shah, A. D.</dc:creator>
<dc:creator>Denaxas, S. C.</dc:creator>
<dc:creator>Hemingway, H.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:date>2018-01-30</dc:date>
<dc:identifier>doi:10.1101/256008</dc:identifier>
<dc:title><![CDATA[Machine learning models in electronic health records can outperform conventional survival models for predicting patient mortality in coronary artery disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/257642v1?rss=1">
<title>
<![CDATA[
ANIMA: Association Network Integration for Multiscale Analysis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/257642v1?rss=1"
</link>
<description><![CDATA[
Contextual functional interpretation of -omics data derived from clinical samples is a classical and difficult problem in computational systems biology. The measurement of thousands of datapoints on single samples has become routine but relating  big data datasets to the complexities of human pathobiology is an area of ongoing research. Complicating this is the fact that many publically available datasets use bulk transcriptomics data from complex tissues like blood. The most prevalent analytic approaches derive molecular  signatures of disease states or apply modular analysis frameworks to the data. Here we show, using a network-based data integration method using clinical phenotype and microarray data as inputs, that we can reconstruct multiple features (or endophenotypes) of disease states at various scales of organization, from transcript abundance patterns of individual genes through co-expression patterns of groups of genes to patterns of cellular behavior in whole blood samples, both in single experiments as well as in a meta-analysis of multiple datasets.
]]></description>
<dc:creator>Deffur, A.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Mayosi, B. M.</dc:creator>
<dc:creator>Mulder, N. M.</dc:creator>
<dc:date>2018-01-31</dc:date>
<dc:identifier>doi:10.1101/257642</dc:identifier>
<dc:title><![CDATA[ANIMA: Association Network Integration for Multiscale Analysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/262501v1?rss=1">
<title>
<![CDATA[
Generative adversarial networks uncover epidermal regulators and predict single cell perturbations 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/262501v1?rss=1"
</link>
<description><![CDATA[
Recent advances have enabled gene expression profiling of single cells at lower cost. As more data is produced there is an increasing need to integrate diverse datasets and better analyse underutilised data to gain biological insights. However, analysis of single cell RNA-seq data is challenging due to biological and technical noise which not only varies between laboratories but also between batches. Here for the first time, we apply a new generative deep learning approach called Generative Adversarial Networks (GAN) to biological data. We apply GANs to epidermal, neural and hematopoietic scRNA-seq data spanning different labs and experimental protocols. We show that it is possible to integrate diverse scRNA-seq datasets and in doing so, our generative model is able to simulate realistic scRNA-seq data that covers the full diversity of cell types. In contrast to many machine-learning approaches, we are able to interpret internal parameters in a biologically meaningful manner. Using our generative model we are able to obtain a universal representation of epidermal differentiation and use this to predict the effect of cell state perturbations on gene expression at high time-resolution. We show that our trained neural networks identify biological state-determining genes and through analysis of these networks we can obtain inferred gene regulatory relationships. Finally, we use internal GAN learned features to perform dimensionality reduction. In combination these attributes provide a powerful framework to progress the analysis of scRNA-seq data beyond exploratory analysis of cell clusters and towards integration of multiple datasets regardless of origin.
]]></description>
<dc:creator>Ghahramani, A.</dc:creator>
<dc:creator>Watt, F. M.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:date>2018-02-08</dc:date>
<dc:identifier>doi:10.1101/262501</dc:identifier>
<dc:title><![CDATA[Generative adversarial networks uncover epidermal regulators and predict single cell perturbations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-02-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/263152v1?rss=1">
<title>
<![CDATA[
TCR-MHC Interaction Strength Defines Trafficking and Resident Memory Status of CD8 T cells in the Brain 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/263152v1?rss=1"
</link>
<description><![CDATA[
T cell receptor-Major histocompatibility complex (TCR-MHC) affinities span a wide range in a polyclonal T cell response, yet it is undefined how affinity shapes long-term properties of CD8 T cells during chronic infection with persistent antigen. Here, we investigate how the affinity of the TCR-MHC interaction shapes the phenotype of memory CD8 T cells in the chronically Toxoplasma gondii-infected brain. We employed CD8 T cells from three lines of transnuclear (TN) mice that harbour in their endogenous loci different T cell receptors specific for the same Toxoplasma antigenic epitope ROP7. The three TN CD8 T cell clones span a wide range of affinities to MHCI-ROP7. These three CD8 T cell clones have a distinct and fixed hierarchy in terms of effector function in response to the antigen measured as proliferation capacity, trafficking, T cell maintenance and memory formation. In particular, the T cell clone of lowest affinity does not home to the brain. The two higher affinity T cell clones show differences in establishing resident memory populations (CD103+) in the brain with the higher affinity clone persisting longer in the host during chronic infection. Transcriptional profiling of naive and activated ROP7-specific CD8 T cells revealed that Klf2 encoding a transcription factor that is known to be a negative marker for T cell trafficking is upregulated in the activated lowest affinity ROP7 clone. Our data thus suggest that TCR-MHC affinity dictates memory CD8 T cell fate at the site of infection.
]]></description>
<dc:creator>Sanecka, A.</dc:creator>
<dc:creator>Yoshida, N.</dc:creator>
<dc:creator>Kolawole, E. M.</dc:creator>
<dc:creator>Patel, H.</dc:creator>
<dc:creator>Evavold, B. D.</dc:creator>
<dc:creator>Frickel, E.</dc:creator>
<dc:date>2018-02-09</dc:date>
<dc:identifier>doi:10.1101/263152</dc:identifier>
<dc:title><![CDATA[TCR-MHC Interaction Strength Defines Trafficking and Resident Memory Status of CD8 T cells in the Brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-02-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/262451v1?rss=1">
<title>
<![CDATA[
Spatio-temporal relays control layer specificity of motion-direction sensitive neurons in Drosophila 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/262451v1?rss=1"
</link>
<description><![CDATA[
Visual motion detection in sighted animals is essential to guide behavioral actions ensuring their survival. In Drosophila, motion direction is first detected by T4/T5 neurons. Their axons innervate one of four lobula plate layers. How T4/T5 neurons with layer-specific representation of motion-direction preferences are specified during development is unknown. We show that diffusible Wingless (Wg) between adjacent neuroepithelia induces its own expression to form secondary signaling centers. These activate Decapentaplegic (Dpp) signaling in adjacent lateral tertiary neuroepithelial domains dedicated to producing layer 3/4-specific T4/T5 neurons. T4/T5 neurons derived from the core domain devoid of Dpp signaling adopt the default layer 1/2 fate. Dpp signaling induces the expression of the T-box transcription factor Optomotor-blind (Omb), serving as a relay to postmitotic neurons. Omb-mediated repression of Dachshund transforms layer 1/2-into layer 3/4-specific neurons. Hence, spatio-temporal relay mechanisms, bridging the distances between neuroepithelial domains and their postmitotic progeny, implement T4/T5 neuron-subtype identity.
]]></description>
<dc:creator>Apitz, H.</dc:creator>
<dc:creator>Salecker, I.</dc:creator>
<dc:date>2018-02-10</dc:date>
<dc:identifier>doi:10.1101/262451</dc:identifier>
<dc:title><![CDATA[Spatio-temporal relays control layer specificity of motion-direction sensitive neurons in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/268037v1?rss=1">
<title>
<![CDATA[
Population Turnover in Remote Oceania Shortly After Initial Settlement 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/268037v1?rss=1"
</link>
<description><![CDATA[
Ancient DNA analysis of three individuals dated to ~3000 years before present (BP) from Vanuatu and one ~2600 BP individual from Tonga has revealed that the first inhabitants of Remote Oceania ("First Remote Oceanians") were almost entirely of East Asian ancestry, and thus their ancestors passed New Guinea, the Bismarck Archipelago, and the Solomon Islands with minimal admixture with the Papuan groups they encountered [1]. However, all present-day populations in Near and Remote Oceania harbor 25-100% Papuan ancestry, implying that there must have been at least one later stream of migration eastward from Near Oceania. We generated genome-wide data for 14 ancient individuals from Efate and Epi Islands in Vanuatu ranging from 3,000-150 BP, along with 185 present-day Vanuatu individuals from 18 islands. We show that people of almost entirely Papuan ancestry had arrived in Vanuatu by 2400 BP, an event that coincided with the end of the Lapita cultural period, changes in skeletal morphology, and the cessation of long-distance trade between Near and Remote Oceania [2]. First Remote Oceanian ancestry subsequently increased via admixture but remains at 10-20% in most islands. Through a fine-grained comparison of ancestry profiles in Vanuatu and Polynesia with diverse groups in Near Oceania, we find that Papuan ancestry in Vanuatu is consistent with deriving from the Bismarck Archipelago instead of the geographically closer Solomon Islands. Papuan ancestry in Polynesia also shows connections to the ancestry profiles present in the Bismarck Archipelago but is more similar to Tolai from New Britain and Tutuba from Vanuatu than to the ancient Vanuatu individuals and the great majority of present-day Vanuatu populations. This suggests a third eastward stream of migration from Near to Remote Oceania bringing a different type of Papuan ancestry.
]]></description>
<dc:creator>Lipson, M.</dc:creator>
<dc:creator>Skoglund, P.</dc:creator>
<dc:creator>Spriggs, M.</dc:creator>
<dc:creator>Valentin, F.</dc:creator>
<dc:creator>Bedford, S.</dc:creator>
<dc:creator>Shing, R.</dc:creator>
<dc:creator>Buckley, H.</dc:creator>
<dc:creator>Phillip, I.</dc:creator>
<dc:creator>Ward, G. K.</dc:creator>
<dc:creator>Mallick, S.</dc:creator>
<dc:creator>Rohland, N.</dc:creator>
<dc:creator>Broomandkhoshbacht, N.</dc:creator>
<dc:creator>Cheronet, O.</dc:creator>
<dc:creator>Ferry, M.</dc:creator>
<dc:creator>Harper, T. K.</dc:creator>
<dc:creator>Michel, M.</dc:creator>
<dc:creator>Oppenheimer, J.</dc:creator>
<dc:creator>Sirak, K.</dc:creator>
<dc:creator>Stewardson, K.</dc:creator>
<dc:creator>Auckland, K.</dc:creator>
<dc:creator>Hill, A. V. S.</dc:creator>
<dc:creator>Maitland, K.</dc:creator>
<dc:creator>Oppenheimer, S. J.</dc:creator>
<dc:creator>Parks, T.</dc:creator>
<dc:creator>Robson, K.</dc:creator>
<dc:creator>Williams, T. N.</dc:creator>
<dc:creator>Kennett, D. J.</dc:creator>
<dc:creator>Mentzer, A. J.</dc:creator>
<dc:creator>Pinhasi, R.</dc:creator>
<dc:creator>Reich, D.</dc:creator>
<dc:date>2018-02-19</dc:date>
<dc:identifier>doi:10.1101/268037</dc:identifier>
<dc:title><![CDATA[Population Turnover in Remote Oceania Shortly After Initial Settlement]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-02-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/271627v1?rss=1">
<title>
<![CDATA[
Differential expression of an alternative splice variant of IL-12Rβ1 impacts early dissemination in the mouse and associates with disease outcome in both mouse and humans exposed to tuberculosis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/271627v1?rss=1"
</link>
<description><![CDATA[
Experimental mouse models of TB suggest that early events in the lung impact immunity. Early events in the human lung in response to TB are difficult to probe and their impact on disease outcome is unknown. We have shown in mouse that a secreted alternatively-spliced variant of IL-12R{beta}1, lacking the transmembrane domain and termed {Delta}TM-IL-12R{beta}1, promotes dendritic cell migration to the draining lymph node, augments T cell activation and limits dissemination of M. tuberculosis (Mtb). We show here that CBA/J and C3H/HeJ mice (both highly susceptible to Mtb) express higher levels of {Delta}TM-IL-12R{beta}1 than resistant C57BL6 mice and limit early dissemination of Mtb from the lungs. Both CD11c+ cells and T cells express {Delta}TM-IL-12R{beta}1 in humans, and mice unable to make {Delta}TM-IL-12R{beta}1 in either CD4 or CD11c expressing cells permit early dissemination from the lung. Analysis of publically available blood transcriptomes indicates that pulmonary TB is associated with high {Delta}TM-IL-12R{beta}1 expression and that of all IL-12 related signals, the {Delta}TM-IL-12R{beta}1 signal best predicts active disease. {Delta}TM-IL-12R{beta}1 expression reflects the heterogeneity of latent TB infection and has the capacity to discriminate between latent and active disease. In a new Chinese TB patient cohort, {Delta}TM-IL-12R{beta}1 effectively differentiates TB from latent TB, healthy controls and pneumonia patients. Finally, {Delta}TM-IL-12R{beta}1 expression drops in drug-treated individuals in the UK and China where infection pressure is low. We propose that {Delta}TM-IL-12R{beta}1 regulates early dissemination from the lung and that it has diagnostic potential and provides mechanistic insights into human TB.
]]></description>
<dc:creator>Das, M. K.</dc:creator>
<dc:creator>Ray, A. A.</dc:creator>
<dc:creator>Cai, Y.</dc:creator>
<dc:creator>Singhania, A.</dc:creator>
<dc:creator>Graham, C.</dc:creator>
<dc:creator>Liao, M.</dc:creator>
<dc:creator>Fountain, J. J.</dc:creator>
<dc:creator>Pearl, J. E.</dc:creator>
<dc:creator>Pareek, M.</dc:creator>
<dc:creator>Haldar, P.</dc:creator>
<dc:creator>O'Garra, A.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Cooper, A. M.</dc:creator>
<dc:date>2018-02-26</dc:date>
<dc:identifier>doi:10.1101/271627</dc:identifier>
<dc:title><![CDATA[Differential expression of an alternative splice variant of IL-12Rβ1 impacts early dissemination in the mouse and associates with disease outcome in both mouse and humans exposed to tuberculosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-02-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/272591v1?rss=1">
<title>
<![CDATA[
Human axial progenitors generate trunk neural crest cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/272591v1?rss=1"
</link>
<description><![CDATA[
The neural crest (NC) is a multipotent embryonic cell population generating distinct cell types in an axial position-dependent manner. The production of NC cells from human pluripotent stem cells (hPSCs) is a valuable approach to study human NC biology. However, the origin of human trunk NC remains undefined and therefore current in vitro differentiation strategies induce only a modest yield of trunk NC cells. Here we show that hPSC-derived axial progenitors, the posteriorly-located drivers of embryonic axis elongation, give rise to trunk NC cells and their derivatives. Moreover, we define the molecular signatures associated with the emergence of human NC cells of distinct axial identities in vitro. Collectively, our findings indicate that there are two routes toward a human post-cranial NC state: the birth of cardiac and vagal NC is facilitated by retinoic acid-induced posteriorisation of an anterior precursor whereas trunk NC arises within a pool of posterior axial progenitors.
]]></description>
<dc:creator>Frith, T. J. R.</dc:creator>
<dc:creator>Granata, I.</dc:creator>
<dc:creator>Stout, E.</dc:creator>
<dc:creator>Wind, M.</dc:creator>
<dc:creator>Thompson, O.</dc:creator>
<dc:creator>Neumann, K.</dc:creator>
<dc:creator>Stavish, D.</dc:creator>
<dc:creator>Heath, P. R.</dc:creator>
<dc:creator>Hackland, J. O. S.</dc:creator>
<dc:creator>Anastassiadis, K.</dc:creator>
<dc:creator>Gouti, M.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Wilson, V.</dc:creator>
<dc:creator>Guarracino, M. R.</dc:creator>
<dc:creator>Andrews, P. W.</dc:creator>
<dc:creator>Tsakiridis, A.</dc:creator>
<dc:date>2018-02-27</dc:date>
<dc:identifier>doi:10.1101/272591</dc:identifier>
<dc:title><![CDATA[Human axial progenitors generate trunk neural crest cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-02-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/275008v1?rss=1">
<title>
<![CDATA[
Predicting improved protein conformations with a temporal deep recurrent neural network 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/275008v1?rss=1"
</link>
<description><![CDATA[
Accurate protein structure prediction from amino acid sequence is still an unsolved problem. The most reliable methods centre on template based modelling. However, the accuracy of these models entirely depends on the availability of experimentally resolved homologous template structures. In order to generate more accurate models, extensive physics based molecular dynamics (MD) refinement simulations are performed to sample many different conformations to find improved conformational states. In this study, we propose a deep recurrent network model, called DeepTrajectory, that is able to identify these improved conformational states, with high precision, from a variety of different MD based sampling protocols. The proposed model learns the temporal patterns of features computed from the MD trajectory data in order to classify whether each recorded simulation snapshot is an improved conformational state, decreased conformational state or a none perceivable change in state with respect to the starting conformation. The model is trained and tested on 904 trajectories from 42 different protein systems with a cumulative number of more than 1.7 million snapshots. We show that our model outperforms other state of the art machine-learning algorithms that do not consider temporal dependencies. To our knowledge, DeepTrajectory is the first implementation of a time-dependent deep-learning protocol that is re-trainable and able to adapt to any new MD based sampling procedure, thereby demonstrating how a neural network can be used to learn the latter part of the protein folding funnel.
]]></description>
<dc:creator>Pfeiffenberger, E.</dc:creator>
<dc:creator>Bates, P. A.</dc:creator>
<dc:date>2018-03-03</dc:date>
<dc:identifier>doi:10.1101/275008</dc:identifier>
<dc:title><![CDATA[Predicting improved protein conformations with a temporal deep recurrent neural network]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/275131v1?rss=1">
<title>
<![CDATA[
A strategy to study intrinsically mixed folded proteins: The structure in solution of ataxin-3 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/275131v1?rss=1"
</link>
<description><![CDATA[
It has increasingly become clear over the last two decades that proteins can contain both globular domains and intrinsically unfolded regions which both can contribute to function. While equally interesting, the disordered regions are difficult to study because they usually do not crystallize unless bound to partners and are not easily amenable to cryo-electron microscopy studies. Nuclear magnetic resonance spectroscopy remains the best technique to capture the structural features of intrinsically mixed folded proteins and describe their dynamics. These studies rely on the successful assignment of the spectrum, task not easy per se given the limited spread of the resonances of the disordered residues. Here, we describe assignment of the spectrum of ataxin-3, the protein responsible for the neurodegenerative Machado-Joseph disease. We used a 42 kDa construct containing a globular N-terminal josephin domain and a C-terminal tail which comprises thirteen polyglutamine repeats within a low-complexity region. We developed a strategy which allowed us to achieve 87% assignment of the spectrum. We show that the C-terminal tail is flexible with extended helical regions and interacts only marginally with the rest of the protein. We could also, for the first time, deduce the structure of the polyglutamine repeats within the context of the full-length protein and show that it has a strong helical propensity stabilized by the preceding region.
]]></description>
<dc:creator>Sicorello, A.</dc:creator>
<dc:creator>Kelly, G.</dc:creator>
<dc:creator>Oregioni, A.</dc:creator>
<dc:creator>Novacek, J.</dc:creator>
<dc:creator>sklenar, v.</dc:creator>
<dc:creator>Pastore, A.</dc:creator>
<dc:date>2018-03-09</dc:date>
<dc:identifier>doi:10.1101/275131</dc:identifier>
<dc:title><![CDATA[A strategy to study intrinsically mixed folded proteins: The structure in solution of ataxin-3]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/281139v1?rss=1">
<title>
<![CDATA[
Neurofilament light as a blood biomarker for neurodegeneration in Down syndrome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/281139v1?rss=1"
</link>
<description><![CDATA[
INTRODUCTIONDown syndrome (DS) may be considered a genetic form of Alzheimers disease (AD) due to universal development of AD neuropathology, but diagnosis and treatment trials are hampered by a lack of reliable blood biomarkers. A potential biomarker is neurofilament light (NF-L), due to its association with axonal damage in neurodegenerative conditions.nnMETHODSWe measured blood NF-L concentration in 100 adults with DS using Simoa NF-light(R) assays, and examined relationships with age, and cross-sectional and longitudinal dementia diagnosis.nnRESULTSNF-L levels increased with age (Spearmans rho = 0.789, p<0.001), with a steep increase after age 40, and were predictive of dementia status (p=0.022 adjusting for age, sex, and APOE4) but showed no relationship with longstanding epilepsy or premorbid ability. Baseline NF-L levels were associated with longitudinal dementia status.nnDISCUSSIONNF-L is a biomarker for neurodegeneration in DS, with potential for use in future clinical trials to prevent or delay dementia.nnResearch in contextO_ST_ABSSystematic reviewC_ST_ABSThe authors reviewed the literature using PubMed searches supplemented with our knowledge of pending papers in this research area. While blood NF-L has been associated with clinical features of progression in a number of neurodegenerative conditions, we have not identified any reports of NF-L associated with cognitive decline in DS, a genetic form of AD.nnInterpretationOur findings demonstrate the potential utility of NF-L as a blood biomarker of neurodegeneration in DS, a population that may not be able to tolerate more invasive procedures such as neuroimaging and lumbar punctures to track progression.nnFuture directionsThe association between NF-L and other markers of longitudinal AD progression should be explored further in future work.
]]></description>
<dc:creator>Strydom, A.</dc:creator>
<dc:creator>Heslegrave, A.</dc:creator>
<dc:creator>Startin, C. M.</dc:creator>
<dc:creator>Mok, K. Y.</dc:creator>
<dc:creator>Hardy, J.</dc:creator>
<dc:creator>Groet, J.</dc:creator>
<dc:creator>Nizetic, D.</dc:creator>
<dc:creator>The LonDownS consortium,</dc:creator>
<dc:creator>Zetterberg, H.</dc:creator>
<dc:date>2018-03-13</dc:date>
<dc:identifier>doi:10.1101/281139</dc:identifier>
<dc:title><![CDATA[Neurofilament light as a blood biomarker for neurodegeneration in Down syndrome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/281519v1?rss=1">
<title>
<![CDATA[
Distinct and stage-specific contributions of TET1 and TET2 to stepwise cytosine oxidation in the transition from naive to primed pluripotency 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/281519v1?rss=1"
</link>
<description><![CDATA[
The TET-oxidized cytosine derivatives, 5-hydroxymethylcytosine (5hmC) and 5-formylcytosine (5fC), are considered DNA demethylation intermediates as well as stable epigenetic marks in mammals. We compared modified cytosine and enzyme levels in TET-knockout cells during naive pluripotency exit and found distinct and differentiation-dependent contributions of TET1 and TET2 to 5hmC and 5fC formation. The divergent modified cytosine levels argue for independent consecutive oxidation steps in vivo with broad implications for epigenetic regulation.
]]></description>
<dc:creator>Mulholland, C. B.</dc:creator>
<dc:creator>Traube, F. R.</dc:creator>
<dc:creator>Parsa, E.</dc:creator>
<dc:creator>Eckl, E.-M.</dc:creator>
<dc:creator>Schoenung, M.</dc:creator>
<dc:creator>Modic, M.</dc:creator>
<dc:creator>Bartoschek, M. D.</dc:creator>
<dc:creator>Stolz, P.</dc:creator>
<dc:creator>Ryan, J.</dc:creator>
<dc:creator>Carell, T.</dc:creator>
<dc:creator>Leonhardt, H.</dc:creator>
<dc:creator>Bultmann, S.</dc:creator>
<dc:date>2018-03-13</dc:date>
<dc:identifier>doi:10.1101/281519</dc:identifier>
<dc:title><![CDATA[Distinct and stage-specific contributions of TET1 and TET2 to stepwise cytosine oxidation in the transition from naive to primed pluripotency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/282699v1?rss=1">
<title>
<![CDATA[
DIA-NN: Deep neural networks substantially improve the identification performance of Data-independent acquisition (DIA) in proteomics 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/282699v1?rss=1"
</link>
<description><![CDATA[
Data-independent acquisition (DIA-MS) strategies, like SWATH-MS, have been developed to increase consistency, quantification precision and proteomic depth in label-free proteomic experiments. They aim to overcome stochasticity in the selection of precursor ions by utilising (mass-) windowed acquisition that is followed by computational reconstruction of the chromatograms. While DIA methods increasingly outperform typical data-dependent methods in identification consistency and precision specifically on large sample series, possibilities remain for further improvements. At present, only a fraction of the information recorded in the complex DIA spectra is extracted by the software analysis pipelines. Here we present a software tool (DIA-NN) that introduces artificial neural nets and a new quantification strategy to enhance signal processing in DIA-data. DIA-NN greatly improves identification of precursor ions and, as a consequence, protein quantification accuracy. The performance of DIA-NN demonstrates that deep learning provides opportunities to boost the analysis of data-independent acquisition workflows in proteomics.
]]></description>
<dc:creator>Demichev, V.</dc:creator>
<dc:creator>Messner, C. B.</dc:creator>
<dc:creator>Lilley, K. S.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:date>2018-03-15</dc:date>
<dc:identifier>doi:10.1101/282699</dc:identifier>
<dc:title><![CDATA[DIA-NN: Deep neural networks substantially improve the identification performance of Data-independent acquisition (DIA) in proteomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/285627v1?rss=1">
<title>
<![CDATA[
Identifying the genetic basis of variation in cell behaviour in human iPS cell lines from healthy donors 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/285627v1?rss=1"
</link>
<description><![CDATA[
Large cohorts of human iPSCs from healthy donors are potentially a powerful tool for investigating the relationship between genetic variants and cellular phenotypes. Here we integrate high content imaging, gene expression and DNA sequence datasets for over 100 human iPSC lines to identify the genetic basis of inter-individual variability in cell behaviour. By applying a dimensionality reduction approach, Probabilistic Estimation of Expression Residuals (PEER), we identified genes that correlated in expression with intrinsic (genetic) and extrinsic (ECM) factors. However, variation in mRNA levels could not account for outlier cell behaviour. Instead, we identified rare, deleterious SNVs in the coding sequence of genes involved in ECM adhesion that occurred in cell lines that were outliers for one or more phenotypes such as cell spreading. These also correlated with altered germ layer differentiation on micropatterned surfaces. Our study thus establishes a strategy for integrating genetic and cell biological measurements for high-throughput analysis.
]]></description>
<dc:creator>Vigilante, A.</dc:creator>
<dc:creator>Laddach, A.</dc:creator>
<dc:creator>Moens, N.</dc:creator>
<dc:creator>Meleckyte, R.</dc:creator>
<dc:creator>Leha, A.</dc:creator>
<dc:creator>Ghahramani, A.</dc:creator>
<dc:creator>Culley, O. J.</dc:creator>
<dc:creator>Kathuria, A.</dc:creator>
<dc:creator>Hurling, C.</dc:creator>
<dc:creator>Vickers, A.</dc:creator>
<dc:creator>Tewary, M.</dc:creator>
<dc:creator>Zandstra, P.</dc:creator>
<dc:creator>HipSci Consortium,</dc:creator>
<dc:creator>Durbin, R.</dc:creator>
<dc:creator>Fraternali, F.</dc:creator>
<dc:creator>Stegle, O.</dc:creator>
<dc:creator>Birney, E.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:creator>Danovi, D.</dc:creator>
<dc:creator>Watt, F. M.</dc:creator>
<dc:date>2018-03-20</dc:date>
<dc:identifier>doi:10.1101/285627</dc:identifier>
<dc:title><![CDATA[Identifying the genetic basis of variation in cell behaviour in human iPS cell lines from healthy donors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/287920v1?rss=1">
<title>
<![CDATA[
Glycosylation-dependent modulation of the lL-2 signaling axis determines Th17 differentiation and IL-10 production 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/287920v1?rss=1"
</link>
<description><![CDATA[
Metabolism plays an essential role in shaping T helper (Th) cell responses including the production of pro-inflammatory cytokines, however the effects on IL-10 have not been investigated. We show that the glucose analogue 2-deoxyglucose (2DG) specifically inhibits Th1 and Th2 cell differentiation and accompanying IL-10 production. In contrast, 2DG promotes IL-17A production by Th17 cells, even in the presence of IL-2 known to limit Th17 differentiation, whilst totally abrogating the production of IL-10. Notably, rather than inhibiting glycolysis, 2DG acts through the inhibition of glycosylation, which is critical for IL-2R surface expression and downstream signaling in both mouse and man. Strikingly, IL-2 is essential for IL-10 production by Th17 cells, in contrast to its inhibitory effect on the production of IL-17A. Our study reveals a previously unappreciated, anti-inflammatory role for IL-2 in Th17 cell production of IL-10 and thus identifies a novel mechanism to limit Th17 pathogenicity.
]]></description>
<dc:creator>Gabrysova, L.</dc:creator>
<dc:creator>Mann, E. H.</dc:creator>
<dc:creator>Bradley, L.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Whicher, C.</dc:creator>
<dc:creator>Hawrylowicz, C. M.</dc:creator>
<dc:creator>Anastasiou, D.</dc:creator>
<dc:creator>O'Garra, A.</dc:creator>
<dc:date>2018-03-23</dc:date>
<dc:identifier>doi:10.1101/287920</dc:identifier>
<dc:title><![CDATA[Glycosylation-dependent modulation of the lL-2 signaling axis determines Th17 differentiation and IL-10 production]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/291815v1?rss=1">
<title>
<![CDATA[
AutonoMouse: High throughput automated operant conditioning shows progressive behavioural impairment with graded olfactory bulb lesions. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/291815v1?rss=1"
</link>
<description><![CDATA[
Operant conditioning is a crucial tool in neuroscience research for probing brain function. While molecular, anatomical and even physiological techniques have seen radical increases in throughput, efficiency, and reproducibility in recent years, behavioural tools have seen much less of an improvement. Here we present a fully automated, high-throughput system for self-initiated conditioning of up to 25 group-housed, radio-frequency identification (RFID) tagged mice over periods of several months and >10^6 trials. We validate this "AutonoMouse" system in a series of olfactory behavioural tasks and show that acquired data is comparable to previous semi-manual approaches. Furthermore, we use AutonoMouse to systematically probe the impact of graded olfactory bulb lesions on olfactory behaviour and resolve the long-standing conundrum about the apparent lack of impact of lesions on olfactory abilities. The modular nature and open-source design of AutonoMouse should allow for similar robust and systematic assessments across neuroscience research areas.
]]></description>
<dc:creator>Erskine, A.</dc:creator>
<dc:creator>Bus, T.</dc:creator>
<dc:creator>Herb, J. T.</dc:creator>
<dc:creator>Schaefer, A. T.</dc:creator>
<dc:date>2018-03-29</dc:date>
<dc:identifier>doi:10.1101/291815</dc:identifier>
<dc:title><![CDATA[AutonoMouse: High throughput automated operant conditioning shows progressive behavioural impairment with graded olfactory bulb lesions.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/287516v1?rss=1">
<title>
<![CDATA[
Comparable affinity of RabGDIα for GTP- and GDP-bound forms of Rab7 supports a four-state transition model for Rab7 subcellular localization 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/287516v1?rss=1"
</link>
<description><![CDATA[
Endolysosomal system is linked to almost all aspects of cell life and diseases, and Rab7 occupies a critical node in this crucial pathway. However, there have been conflicting views about the exact role of Rab7 in membrane trafficking, since some studies have reported that Rab7 regulates the trafficking from early to late endosomes, while others highlighted its role in late endosomes to lysosomes progression. In the present study, we have revisited this issue from a new viewpoint. In COS-7 cells, a GDP-bound Rab7 mutant, T22N, was located to vesicular membranes as well as in cytoplasm. Similarly, the GTPase-deficient Q67L mutant of Rab7 resided in cytoplasm as well as on membranes. Additionally, we found that RabGDI interacted with both GTP- and GDP-bound forms of Rab7 in vitro. These results have prompted us to propose a four-state transition model for Rab7. This four-state model matches with our recent findings that Rab7 was initially recruited to macropinosomes in a GDP-bound inactive form and subsequently became activated during endocytic maturation in EGF-stimulated COS-7 cells.
]]></description>
<dc:creator>Kanemitsu, A. F.</dc:creator>
<dc:creator>Morishita, S.</dc:creator>
<dc:creator>Kjaer, S.</dc:creator>
<dc:creator>Fukuda, M.</dc:creator>
<dc:creator>Schiavo, G.</dc:creator>
<dc:creator>Nakamura, T.</dc:creator>
<dc:date>2018-03-27</dc:date>
<dc:identifier>doi:10.1101/287516</dc:identifier>
<dc:title><![CDATA[Comparable affinity of RabGDIα for GTP- and GDP-bound forms of Rab7 supports a four-state transition model for Rab7 subcellular localization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/293951v1?rss=1">
<title>
<![CDATA[
A non-cell autonomous actin redistribution enables isotropic retinal growth 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/293951v1?rss=1"
</link>
<description><![CDATA[
Tissue shape is often established early in development and needs to be scaled isotropically during growth. However, the cellular contributors and ways in which cells interact inside tissues to enable coordinated isotropic tissue scaling are not yet understood. Here, we follow cell and tissue shape changes in the zebrafish retinal neuroepithelium, which forms a cup with a smooth surface early in development and maintains this architecture as it grows. By combining 3D analysis and theory, we show that a global increase in cell height is necessary to maintain this tissue shape during growth. Timely cell height increase is governed by non-cell autonomous actin redistribution. Blocking actin redistribution and cell height increase perturbs isotropic scaling and leads to disturbed, folded tissue shape. Taken together, our data show how global changes in cell shape enable isotropic growth of the developing retinal neuroepithelium, a concept that could also apply to other systems.
]]></description>
<dc:creator>Matejcic, M.</dc:creator>
<dc:creator>Salbreux, G.</dc:creator>
<dc:creator>Norden, C.</dc:creator>
<dc:date>2018-04-03</dc:date>
<dc:identifier>doi:10.1101/293951</dc:identifier>
<dc:title><![CDATA[A non-cell autonomous actin redistribution enables isotropic retinal growth]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/297853v1?rss=1">
<title>
<![CDATA[
Heteromeric RNP assembly at LINEs controls lineage-specific RNA processing 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/297853v1?rss=1"
</link>
<description><![CDATA[
It is challenging for RNA processing machineries to select exons within long intronic regions. We find that intronic LINE repeat sequences (LINEs) contribute to this selection by recruiting dozens of RNA-binding proteins (RBPs). This includes MATR3, which promotes binding of PTBP1 to multivalent binding sites in LINEs. Both RBPs repress splicing and 3 end processing within and around LINEs, as demonstrated in cultured human cells and mouse brain. Notably, repressive RBPs preferentially bind to evolutionarily young LINEs, which are confined to deep intronic regions. These RBPs insulate both LINEs and surrounding regions from RNA processing. Upon evolutionary divergence, gradual loss of insulation diversifies the roles of LINEs. Older LINEs are located closer to exons, are a common source of tissue-specific exons, and increasingly bind to RBPs that enhance RNA processing. Thus, LINEs are hubs for assembly of repressive RBPs, and contribute to evolution of new, lineage-specific transcripts in mammals.
]]></description>
<dc:creator>Attig, J.</dc:creator>
<dc:creator>Agostini, F.</dc:creator>
<dc:creator>Gooding, C.</dc:creator>
<dc:creator>Singh, A.</dc:creator>
<dc:creator>Chakrabarti, A. M.</dc:creator>
<dc:creator>Haberman, N.</dc:creator>
<dc:creator>Emmett, W.</dc:creator>
<dc:creator>Smith, C. W.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:date>2018-04-09</dc:date>
<dc:identifier>doi:10.1101/297853</dc:identifier>
<dc:title><![CDATA[Heteromeric RNP assembly at LINEs controls lineage-specific RNA processing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/306803v1?rss=1">
<title>
<![CDATA[
Maternal pluripotency factors prime the zygotic genome to respond to intercellular signals 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/306803v1?rss=1"
</link>
<description><![CDATA[
Embryonic development yields many different cell types in response to just a few families of inductive signals. The property of a signal-receiving cell that determines how it responds to such signals, including the activation of cell type-specific genes, is known as its competence. Here, we show how maternal factors modify chromatin to specify initial competence in the frog Xenopus tropicalis. We identified the earliest engaged regulatory DNA sequences, and inferred from them critical activators of the zygotic genome. Of these, we showed that the pioneering activity of the maternal pluripotency factors Pou5f3 and Sox3 predefines competence for germ layer formation by extensively remodeling compacted chromatin before the onset of signaling. The remodeling includes the opening and marking of thousands of regulatory elements, extensive chromatin looping, and the co-recruitment of signal-mediating transcription factors. Our work identifies significant developmental principles that inform our understanding of how pluripotent stem cells interpret inductive signals.
]]></description>
<dc:creator>Gentsch, G. E.</dc:creator>
<dc:creator>Spruce, T.</dc:creator>
<dc:creator>Owens, N. D. L.</dc:creator>
<dc:creator>Smith, J. C.</dc:creator>
<dc:date>2018-04-23</dc:date>
<dc:identifier>doi:10.1101/306803</dc:identifier>
<dc:title><![CDATA[Maternal pluripotency factors prime the zygotic genome to respond to intercellular signals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/310425v1?rss=1">
<title>
<![CDATA[
Creating Standards for Evaluating Tumour Subclonal Reconstruction 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/310425v1?rss=1"
</link>
<description><![CDATA[
Tumours evolve through time and space. Computational techniques have been developed to infer their evolutionary dynamics from DNA sequencing data. A growing number of studies have used these approaches to link molecular cancer evolution to clinical progression and response to therapy. There has not yet been a systematic evaluation of methods for reconstructing tumour subclonality, in part due to the underlying mathematical and biological complexity and to difficulties in creating gold-standards. To fill this gap, we systematically elucidated the key algorithmic problems in subclonal reconstruction and developed mathematically valid quantitative metrics for evaluating them. We then created approaches to simulate realistic tumour genomes, harbouring all known mutation types and processes both clonally and subclonally. We then simulated 580 tumour genomes for reconstruction, varying tumour read-depth and benchmarking somatic variant detection and subclonal reconstruction strategies. The inference of tumour phylogenies is rapidly becoming standard practice in cancer genome analysis; this study creates a baseline for its evaluation.
]]></description>
<dc:creator>Boutros, P. C.</dc:creator>
<dc:creator>Salcedo, A.</dc:creator>
<dc:creator>Tarabichi, M.</dc:creator>
<dc:creator>Espiritu, S. M. G.</dc:creator>
<dc:creator>Deshwar, A. G.</dc:creator>
<dc:creator>David, M.</dc:creator>
<dc:creator>Wilson, N. M.</dc:creator>
<dc:creator>Dentro, S.</dc:creator>
<dc:creator>Wintersinger, J. A.</dc:creator>
<dc:creator>Liu, L. Y.</dc:creator>
<dc:creator>Ko, M.</dc:creator>
<dc:creator>Sivanandan, S.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Zhu, K.</dc:creator>
<dc:creator>Yang, T.-H.</dc:creator>
<dc:creator>Chilton, J. M.</dc:creator>
<dc:creator>Buchanan, A.</dc:creator>
<dc:creator>Lalansingh, C. M.</dc:creator>
<dc:creator>P'ng, C.</dc:creator>
<dc:creator>Anghel, C. V.</dc:creator>
<dc:creator>Umar, I.</dc:creator>
<dc:creator>Lo, B.</dc:creator>
<dc:creator>Simpson, J. T.</dc:creator>
<dc:creator>Stuart, J. M.</dc:creator>
<dc:creator>Anastassiou, D.</dc:creator>
<dc:creator>Guan, Y.</dc:creator>
<dc:creator>Ewing, A.</dc:creator>
<dc:creator>Ellrott, K.</dc:creator>
<dc:creator>Wedge, D. C.</dc:creator>
<dc:creator>Morris, Q. D.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>DREAM SMC-Het Participants,</dc:creator>
<dc:date>2018-04-28</dc:date>
<dc:identifier>doi:10.1101/310425</dc:identifier>
<dc:title><![CDATA[Creating Standards for Evaluating Tumour Subclonal Reconstruction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/253203v1?rss=1">
<title>
<![CDATA[
Modelling mitosis with multiple phenotypes: relation to Haeckels biogenetic recapitulation law 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/253203v1?rss=1"
</link>
<description><![CDATA[
The article presents a novel stochastic mathematical model of mitosis in heterogeneous (multiple-phenotype), age-dependent cell populations. The developed computational techniques involve flexible use of differentiation tree diagrams. The applicability of the model is discussed in the context of the Haeckelian (biogenetic) paradigm. In particular, the article puts forward the conjecture of generality of Haeckels recapitulation law. The conjecture is briefly collated against relevant scientific evidence and elaborated for the specific case of evolving/mutable cell phenotypes as considered by the model. The feasibility, basic regimes and the convenience of the model are tested on examples and experimental data, and the corresponding open source simulation software is described and demonstrated.
]]></description>
<dc:creator>Alexandrov, Y.</dc:creator>
<dc:date>2018-01-24</dc:date>
<dc:identifier>doi:10.1101/253203</dc:identifier>
<dc:title><![CDATA[Modelling mitosis with multiple phenotypes: relation to Haeckels biogenetic recapitulation law]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/314559v1?rss=1">
<title>
<![CDATA[
Genomic determinants of sympatric speciation of the Mycobacterium tuberculosis complex across evolutionary timescales. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/314559v1?rss=1"
</link>
<description><![CDATA[
BACKGROUNDModels on how bacterial lineages differentiate increase our understanding on early bacterial speciation events and about the genetic loci involved. Here, we analyze the population genomics events leading to the emergence of the tuberculosis pathogen.nnRESULTSThe emergence is characterized by a combination of recombination events involving core pathogenesis functions and purifying selection on early diverging loci. We identify the phoR gene, the sensor kinase of a two-component system involved in virulence, as a key functional player subject to pervasive positive selection after the divergence of the MTBC from its ancestor. Previous evidence showed that phoR mutations played a central role in the adaptation of the pathogen to different host species. Now we show that phoR have been under selection during the early spread of human tuberculosis, during later expansions and in on-going transmission events.nnCONCLUSIONSOur results show that linking pathogen evolution across evolutionary and epidemiological timescales point to past and present virulence determinants.
]]></description>
<dc:creator>Chiner-Oms, A.</dc:creator>
<dc:creator>Sanchez-Buso, L.</dc:creator>
<dc:creator>Corander, J.</dc:creator>
<dc:creator>Gagneux, S.</dc:creator>
<dc:creator>Harris, S.</dc:creator>
<dc:creator>Young, D.</dc:creator>
<dc:creator>Gonzalez-Candelas, F.</dc:creator>
<dc:creator>Comas, I.</dc:creator>
<dc:date>2018-05-04</dc:date>
<dc:identifier>doi:10.1101/314559</dc:identifier>
<dc:title><![CDATA[Genomic determinants of sympatric speciation of the Mycobacterium tuberculosis complex across evolutionary timescales.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/312041v1?rss=1">
<title>
<![CDATA[
Portraits of genetic intra-tumour heterogeneity and subclonal selection across cancer types 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/312041v1?rss=1"
</link>
<description><![CDATA[
Intra-tumor heterogeneity (ITH) is a mechanism of therapeutic resistance and therefore an important clinical challenge. However, the extent, origin and drivers of ITH across cancer types are poorly understood. To address this question, we extensively characterize ITH across whole-genome sequences of 2,658 cancer samples, spanning 38 cancer types. Nearly all informative samples (95.1%) contain evidence of distinct subclonal expansions, with frequent branching relationships between subclones. We observe positive selection of subclonal driver mutations across most cancer types, and identify cancer type specific subclonal patterns of driver gene mutations, fusions, structural variants and copy-number alterations, as well as dynamic changes in mutational processes between subclonal expansions. Our results underline the importance of ITH and its drivers in tumor evolution, and provide an unprecedented pan-cancer resource of comprehensively annotated subclonal events from whole-genome sequencing data.
]]></description>
<dc:creator>Dentro, S. C.</dc:creator>
<dc:creator>Leshchiner, I.</dc:creator>
<dc:creator>Haase, K.</dc:creator>
<dc:creator>Tarabichi, M.</dc:creator>
<dc:creator>Wintersinger, J.</dc:creator>
<dc:creator>Deshwar, A. G.</dc:creator>
<dc:creator>Yu, K.</dc:creator>
<dc:creator>Rubanova, Y.</dc:creator>
<dc:creator>Mcintyre, G.</dc:creator>
<dc:creator>Vazquez-Garcia, I.</dc:creator>
<dc:creator>Kleinheinz, K.</dc:creator>
<dc:creator>Livitz, D. G.</dc:creator>
<dc:creator>Malikic, S.</dc:creator>
<dc:creator>Donmez, N.</dc:creator>
<dc:creator>Sengupta, S.</dc:creator>
<dc:creator>Demeulemeester, J.</dc:creator>
<dc:creator>Anur, P.</dc:creator>
<dc:creator>Jolly, C.</dc:creator>
<dc:creator>Cmero, M.</dc:creator>
<dc:creator>Rosebrock, D.</dc:creator>
<dc:creator>Schumacher, S.</dc:creator>
<dc:creator>Fan, Y.</dc:creator>
<dc:creator>Fittall, M.</dc:creator>
<dc:creator>Drews, R. M.</dc:creator>
<dc:creator>Yao, X.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Schlesner, M.</dc:creator>
<dc:creator>Adams, D. J.</dc:creator>
<dc:creator>Getz, G.</dc:creator>
<dc:creator>Boutros, P. C.</dc:creator>
<dc:creator>Imielinski, M.</dc:creator>
<dc:creator>Beroukhim, R.</dc:creator>
<dc:creator>Sahinalp, S. C.</dc:creator>
<dc:creator>Ji, Y.</dc:creator>
<dc:creator>Peifer, M.</dc:creator>
<dc:creator>Martincorena, I.</dc:creator>
<dc:creator>Markowetz, F.</dc:creator>
<dc:creator>Mustonen, V.</dc:creator>
<dc:creator>Yuan, K.</dc:creator>
<dc:creator>Gerstung, M.</dc:creator>
<dc:creator>Spellman, P. T.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Morris, Q. D.</dc:creator>
<dc:creator>Wedge, D.</dc:creator>
<dc:date>2018-05-05</dc:date>
<dc:identifier>doi:10.1101/312041</dc:identifier>
<dc:title><![CDATA[Portraits of genetic intra-tumour heterogeneity and subclonal selection across cancer types]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/320416v1?rss=1">
<title>
<![CDATA[
Automating multimodal microscopy with NanoJ-Fluidics 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/320416v1?rss=1"
</link>
<description><![CDATA[
Fluorescence microscopy can reveal all aspects of cellular mechanisms, from molecular details to dynamics, thanks to approaches such as super-resolution and live-cell imaging. Each of its modalities requires specific sample preparation and imaging conditions to obtain high-quality, artefact-free images, ultimately providing complementary information. Combining and multiplexing microscopy approaches is crucial to understand cellular events, but requires elaborate workflows involving multiple sample preparation steps. We present a robust fluidics approach to automate complex sequences of treatment, labelling and imaging of live and fixed cells. Our open-source NanoJ-Fluidics system is based on low-cost LEGO hardware controlled by ImageJ-based software and can be directly adapted to any microscope, providing easy-to-implement high-content, multimodal imaging with high reproducibility. We demonstrate its capacity to carry out complex sequences of experiments such as super-resolved live-to-fixed imaging to study actin dynamics; highly-multiplexed STORM and DNA-PAINT acquisitions of multiple targets; and event-driven fixation microscopy to study the role of adhesion contacts in mitosis.
]]></description>
<dc:creator>Almada, P.</dc:creator>
<dc:creator>Pereira, P.</dc:creator>
<dc:creator>Culley, S.</dc:creator>
<dc:creator>Caillol, G.</dc:creator>
<dc:creator>Boroni-Rueda, F.</dc:creator>
<dc:creator>Dix, C. L.</dc:creator>
<dc:creator>Laine, R. F.</dc:creator>
<dc:creator>Charras, G.</dc:creator>
<dc:creator>Baum, B.</dc:creator>
<dc:creator>Leterrier, C.</dc:creator>
<dc:creator>Henriques, R.</dc:creator>
<dc:date>2018-05-14</dc:date>
<dc:identifier>doi:10.1101/320416</dc:identifier>
<dc:title><![CDATA[Automating multimodal microscopy with NanoJ-Fluidics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/321612v1?rss=1">
<title>
<![CDATA[
Patient-specific detection of cancer genes reveals recurrently perturbed processes in esophageal adenocarcinoma 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/321612v1?rss=1"
</link>
<description><![CDATA[
The identification of somatic alterations with a cancer promoting role is challenging in highly unstable and heterogeneous cancers, such as esophageal adenocarcinoma (EAC). Here we developed a machine learning algorithm to identify cancer genes in individual patients considering all types of damaging alterations simultaneously (mutations, copy number alterations and structural rearrangements). Analysing 261 EACs from the OCCAMS Consortium, we discovered a large number of novel cancer genes that, together with well-known drivers, help promote cancer. Validation using 107 additional EACs confirmed the robustness of the approach. Unlike known drivers whose alterations recur across patients, the large majority of the newly discovered cancer genes are rare or patient-specific. Despite this, they converge towards perturbing cancer-related processes, including intracellular signalling, cell cycle regulation, proteasome activity and Toll-like receptor signalling. Recurrence of process perturbation, rather than individual genes, divides EACs into six clusters that differ in their molecular and clinical features and suggest patient stratifications for personalised treatments. By experimentally mimicking or reverting alterations of predicted cancer genes, we validated their contribution to cancer progression and revealed EAC acquired dependencies, thus demonstrating their potential as therapeutic targets.
]]></description>
<dc:creator>Mourikis, T.</dc:creator>
<dc:creator>Benedetti, L.</dc:creator>
<dc:creator>Foxall, E.</dc:creator>
<dc:creator>Perner, J.</dc:creator>
<dc:creator>Cereda, M.</dc:creator>
<dc:creator>Lagergren, J.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>Yau, C.</dc:creator>
<dc:creator>Fitzgerald, R.</dc:creator>
<dc:creator>Scaffidi, P.</dc:creator>
<dc:creator>Ciccarelli, F. D.</dc:creator>
<dc:date>2018-05-14</dc:date>
<dc:identifier>doi:10.1101/321612</dc:identifier>
<dc:title><![CDATA[Patient-specific detection of cancer genes reveals recurrently perturbed processes in esophageal adenocarcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/308353v1?rss=1">
<title>
<![CDATA[
Mechanics of CaMKII-actin networks 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/308353v1?rss=1"
</link>
<description><![CDATA[
Calcium calmodulin dependent kinase (CaMKII) has an important role in dendritic spine remodelling upon synaptic stimulation. Using fluorescence video microscopy and image analysis, we investigated the architectural dynamics of rhodamine-phalloidin stabilized F-actin networks cross-linked by CaMKII. We used automated image analysis to identify F-actin bundles and cross-over junctions and developed a dimensionless metric to characterize network architecture. Similar networks were formed by three different CaMKII species with ten-fold length difference in the linker region between the kinase domain and holoenzyme hub; implying linker length is not a primary determinant of F-actin binding. Electron micrographs showed that, at physiological molar ratios, single CaMKII holoenzymes cross-linked multiple F-actin filaments in random networks, whereas at higher CaMKII / F-actin ratios filaments bundled. Light microscopy established that random networks resisted macromolecular crowding, with polyethylene glycol mimicking cytoplasmic osmolarity, and blocked ATP-powered compaction by myosin-2 mini-filaments. Importantly, the networks disassembled following addition of calcium calmodulin and were then rapidly spaced into compacted foci by myosin motors or, more slowly, aggregated by crowding. Single molecule TIRF microscopy showed CaMKII dissociation from surface-immobilized G-actin exhibited a mono-exponential dwell-time distribution, whereas CaMKII bound to F-actin networks had a long-lived fraction, trapped at cross-over junctions. Release of CaMKII from F-actin, triggered by calcium calmodulin did not require ATP (hence phosphorylation) and was too rapid to measure with video-rate imaging. The residual bound-fraction was reduced substantially upon addition of an NMDA receptor peptide analogue. These results provide mechanistic insights to CaMKII-actin interactions at the collective network and single molecule level. Our findings argue that CaMKII-actin networks in dendritic spines are stable enough to protect the basal network architecture against physical stress but once CaMKII is disengaged by calcium calmodulin and sequestered by receptors at the synapse; F-actin compaction by myosin motors stabilizes the expanded spine compatible with the recorded times.
]]></description>
<dc:creator>Khan, S. M.</dc:creator>
<dc:creator>Molloy, J. E.</dc:creator>
<dc:creator>Downing, K. H.</dc:creator>
<dc:date>2018-05-14</dc:date>
<dc:identifier>doi:10.1101/308353</dc:identifier>
<dc:title><![CDATA[Mechanics of CaMKII-actin networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/322404v1?rss=1">
<title>
<![CDATA[
Discovery of a novel stereospecific β-hydroxyacyl-CoA lyase/thioesterase shared by three metabolic pathways in Mycobacterium tuberculosis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/322404v1?rss=1"
</link>
<description><![CDATA[
The vast number of poorly characterised enzymes in Mycobacterium tuberculosis (Mtb) is one of the key barriers precluding a better understanding of the biology that underpins pathogenesis. Here, we investigated the Mtb orphan enzyme Rv2498c to delineate its physiological role. Our results from in vitro enzymatic assays, phylogenetic analysis, X-ray crystallography and in vivo Mtb experiments, de-orphan Rv2498c as a multi-functional {beta}-hydroxyacyl-CoA lyase/thioesterase ({beta}-HAClyase/thioesterase) that participates in three different metabolic pathways: L-leucine catabolism, itaconate dissimilation, and glyoxylate shunt. Moreover, the deletion of the rv2498c gene from the Mtb genome resulted in attenuation in the mouse model compared to infection with the parent strain. To the best of our knowledge, this is the first report of an (R)-3-hydroxyl-3-methylglutaryl-CoA for leucine catabolism and an itaconate-specific resistance mechanism in Mtb.
]]></description>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Fedorov, A. A.</dc:creator>
<dc:creator>Fedorov, E. V.</dc:creator>
<dc:creator>Hunt, D. M.</dc:creator>
<dc:creator>Rodgers, A.</dc:creator>
<dc:creator>Garza-Garcia, A.</dc:creator>
<dc:creator>Bonanno, J. B.</dc:creator>
<dc:creator>Almo, S. C.</dc:creator>
<dc:creator>Carvalho, L. P.</dc:creator>
<dc:date>2018-05-15</dc:date>
<dc:identifier>doi:10.1101/322404</dc:identifier>
<dc:title><![CDATA[Discovery of a novel stereospecific β-hydroxyacyl-CoA lyase/thioesterase shared by three metabolic pathways in Mycobacterium tuberculosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/321604v1?rss=1">
<title>
<![CDATA[
TET1 drives global DNA demethylation via DPPA3-mediated inhibition of maintenance methylation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/321604v1?rss=1"
</link>
<description><![CDATA[
Genome-wide DNA demethylation is a unique feature of mammalian development and naive pluripotent stem cells. So far, it was unclear how mammals specifically achieve global DNA hypomethylation, given the high conservation of the DNA (de-)methylation machinery among vertebrates. We found that DNA demethylation requires TET activity but mostly occurs at sites where TET proteins are not bound suggesting a rather indirect mechanism. Among the few specific genes bound and activated by TET proteins was the naive pluripotency and germline marker Dppa3 (Pgc7, Stella), which undergoes TDG dependent demethylation. The requirement of TET proteins for genome-wide DNA demethylation could be bypassed by ectopic expression of Dppa3. We show that DPPA3 binds and displaces UHRF1 from chromatin and thereby prevents the recruitment and activation of the maintenance DNA methyltransferase DNMT1. We demonstrate that DPPA3 alone can drive global DNA demethylation when transferred to amphibians (Xenopus) and fish (medaka), both species that naturally do not have a Dppa3 gene and exhibit no post-fertilization DNA demethylation. Our results show that TET proteins are responsible for active and - indirectly also for - passive DNA demethylation; while TET proteins initiate local and gene-specific demethylation in vertebrates, the recent emergence of DPPA3 introduced a unique means of genome-wide passive demethylation in mammals and contributed to the evolution of epigenetic regulation during early mammalian development.
]]></description>
<dc:creator>Mulholland, C. B.</dc:creator>
<dc:creator>Ryan, J.</dc:creator>
<dc:creator>Qin, W.</dc:creator>
<dc:creator>Bartoschek, M. D.</dc:creator>
<dc:creator>Traube, F. R.</dc:creator>
<dc:creator>Parsa, E.</dc:creator>
<dc:creator>Modic, M.</dc:creator>
<dc:creator>Nixdorf, D.</dc:creator>
<dc:creator>Ziegenhain, C.</dc:creator>
<dc:creator>Carell, T.</dc:creator>
<dc:creator>Enard, W.</dc:creator>
<dc:creator>Bultmann, S.</dc:creator>
<dc:creator>Leonhardt, H.</dc:creator>
<dc:date>2018-05-15</dc:date>
<dc:identifier>doi:10.1101/321604</dc:identifier>
<dc:title><![CDATA[TET1 drives global DNA demethylation via DPPA3-mediated inhibition of maintenance methylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/314310v1?rss=1">
<title>
<![CDATA[
Repression of divergent noncoding transcription by a sequence-specific transcription factor 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/314310v1?rss=1"
</link>
<description><![CDATA[
Many active eukaryotic gene promoters exhibit divergent noncoding transcription, but the mechanisms restricting expression of these transcripts are not well understood. Here we demonstrate how a sequence-specific transcription factor represses divergent noncoding transcription at highly expressed genes in yeast. We find that depletion of the transcription factor Rap1 induces noncoding transcription in a large fraction of Rap1 regulated gene promoters. Specifically, Rap1 prevents transcription initiation at cryptic promoters near its binding sites, which is uncoupled from transcription regulation in the protein-coding direction. We further provide evidence that Rap1 acts independently of chromatin-based mechanisms to repress cryptic or divergent transcription. Finally, we show that divergent transcription in the absence of Rap1 is elicited by the RSC chromatin remodeller. We propose that a sequence-specific transcription factor limits access of basal transcription machinery to regulatory elements and adjacent sequences that act as divergent cryptic promoters, thereby providing directionality towards productive transcription.
]]></description>
<dc:creator>Wu, A. C.</dc:creator>
<dc:creator>Patel, H.</dc:creator>
<dc:creator>Chia, M.</dc:creator>
<dc:creator>Moretto, F.</dc:creator>
<dc:creator>Frith, D.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>van Werven, F. J.</dc:creator>
<dc:date>2018-05-17</dc:date>
<dc:identifier>doi:10.1101/314310</dc:identifier>
<dc:title><![CDATA[Repression of divergent noncoding transcription by a sequence-specific transcription factor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/339671v1?rss=1">
<title>
<![CDATA[
Phosphorylation of a Toxoplasma gondii tyrosine transporter by calcium-dependent kinase 3 is important for parasite fitness 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/339671v1?rss=1"
</link>
<description><![CDATA[
Toxoplasma gondii parasites rapidly exit their host cell when exposed to calcium ionophores. The calcium-dependent protein kinase 3 (TgCDPK3) was previously identified as a key mediator in this process, as TgCDPK3 knockout ({Delta}cdpk3) parasites fail to egress in a timely manner. Phosphoproteomic analysis comparing WT with {Delta}cdpk3 parasites revealed changes in the TgCDPK3-dependent phosphoproteome that included proteins important for regulating motility, but also metabolic enzymes, indicating that TgCDPK3 controls processes beyond egress. Here we have investigated a predicted direct target of TgCDPK3, a putative transporter of the major facilitator superfamily (MFS) and show that it is rapidly phosphorylated after induction of calcium signalling. Conditional knockout (KO) of the transporter reveals an essential role in the lytic cycle during intracellular growth with a transcriptome signature of amino acid-starved parasites. Using a combination of metabolomics and heterologous expression, we confirmed a primary role in tyrosine import. Complementation with phosphorylation site mutants shows that phosphorylation of serine 56 (S56) by TgCDPK3 gives the parasites a growth benefit in competition assays. Collectively, these findings validate an important, albeit non-essential role for TgCDPK3 in the regulation of metabolic processes, in addition to motility.nnAuthor summaryToxoplasma gondii is an obligate intracellular parasite. To survive and spread throughout the host it must repeatedly infect, replicate within and exit, host cells. These recurring cycles of infection and egress rely on signalling pathways that allow the parasites to sense and respond rapidly to their environment. While some key kinases and secondary messengers within these pathways have been identified, functional analysis of non-kinases has been very limited. This is especially true for candidates that are not predicted to play a role in active motility or are not known to function in established signalling pathways. Here we have followed up on an unexpected target of the T. gondii calcium-dependent kinase 3 (TgCDPK3), a plant-like calcium dependent kinase, that was previously shown to play an important role in calcium-mediated exit from the host cell. We show that, in addition to controlling motility of the parasite (as previously shown), TgCDPK3 phosphorylates an essential tyrosine transporter in the plasma membrane. Mutational analysis of the phosphorylation sites demonstrates an important role in maintaining parasite fitness, thus demonstrating that TgCDPK3 plays a pleiotropic role in controlling both egress and metabolism.
]]></description>
<dc:creator>Wallbank, B. A.</dc:creator>
<dc:creator>Dominicus, C. S.</dc:creator>
<dc:creator>Broncel, M.</dc:creator>
<dc:creator>Legrave, N.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Staines, H. M.</dc:creator>
<dc:creator>Treeck, M.</dc:creator>
<dc:date>2018-06-05</dc:date>
<dc:identifier>doi:10.1101/339671</dc:identifier>
<dc:title><![CDATA[Phosphorylation of a Toxoplasma gondii tyrosine transporter by calcium-dependent kinase 3 is important for parasite fitness]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/344986v1?rss=1">
<title>
<![CDATA[
Modelling the nuclear envelope of HeLa cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/344986v1?rss=1"
</link>
<description><![CDATA[
This paper describes a framework for the automatic segmentation of the nuclear envelope of cancerous HeLa cells and the modelling of the volumetric shape against an ellipsoid. The framework is automatic and unsupervised and reported a Jaccard Similarity Index of 0.968 against a manual segmentation. The modelling of the surface provides a visual display of the variations, both smooth and rugged over the surface, and measurements can be extracted with the expectation that they can correlate with the biological characteristics of the cells.
]]></description>
<dc:creator>Karabag, C.</dc:creator>
<dc:creator>Jones, M. L.</dc:creator>
<dc:creator>Peddie, C. J.</dc:creator>
<dc:creator>Weston, A. E.</dc:creator>
<dc:creator>Collinson, L. M.</dc:creator>
<dc:creator>Reyes-Aldasoro, C. C.</dc:creator>
<dc:date>2018-06-12</dc:date>
<dc:identifier>doi:10.1101/344986</dc:identifier>
<dc:title><![CDATA[Modelling the nuclear envelope of HeLa cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/350819v1?rss=1">
<title>
<![CDATA[
Plasmodium-specific atypical memory B cells are not part of the long-lived memory response 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/350819v1?rss=1"
</link>
<description><![CDATA[
A subset of atypical memory B cells accumulates in malaria and several infections, autoimmune disorders and aging in both humans and mice. It has been suggested these cells are exhausted long-lived memory B cells, and their accumulation may contribute to poor acquisition of long-lasting immunity to certain chronic infections, such as malaria and HIV. Here, we generated an immunoglobulin heavy chain knock-in mouse with a BCR that recognizes MSP1 of the rodent malaria parasite, Plasmodium chabaudi. In combination with a mosquito-initiated P. chabaudi infection, we show that Plasmodium-specific atypical memory B cells are short-lived and disappear upon natural resolution of chronic infection. These cells show features of activation, proliferation, DNA replication, and plasmablasts. Our data demonstrate that Plasmodium-specific atypical memory B cells are not a subset of long-lived memory B cells, but rather short-lived activated cells, and part of a physiologic ongoing B-cell response.
]]></description>
<dc:creator>Perez-Mazliah, D.</dc:creator>
<dc:creator>Gardner, P. J.</dc:creator>
<dc:creator>Schweighoffer, E.</dc:creator>
<dc:creator>McLaughlin, S.</dc:creator>
<dc:creator>Hosking, C.</dc:creator>
<dc:creator>Tumwine, I.</dc:creator>
<dc:creator>Davis, R. S.</dc:creator>
<dc:creator>Potocnik, A. J.</dc:creator>
<dc:creator>Tybulewicz, V.</dc:creator>
<dc:creator>langhorne, j.</dc:creator>
<dc:date>2018-06-22</dc:date>
<dc:identifier>doi:10.1101/350819</dc:identifier>
<dc:title><![CDATA[Plasmodium-specific atypical memory B cells are not part of the long-lived memory response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/354407v1?rss=1">
<title>
<![CDATA[
Dual RNAseq of human leprosy lesions identifies bacterial determinants linked to host immune response 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/354407v1?rss=1"
</link>
<description><![CDATA[
To understand how the interaction between an intracellular bacterium and the host immune system contributes to outcome at the site of infection, we studied leprosy, a disease that forms a clinical spectrum, in which progressive infection by the intracellular bacterium Mycobacterium leprae is characterized by the production of type | IFNs and antibody production. We performed dual RNAseq on patient lesions, identifying a continuum of distinct bacterial states that are linked to the host immune response. The bacterial burden, represented by the fraction of bacterial transcripts, correlates with a host type | IFN gene signature, known to inhibit antimicrobial responses. Second, the bacterial transcriptional activity, defined by the bacterial mRNA/rRNA ratio, links bacterial heat shock proteins with the BAFF-BCMA host antibody response pathway. Our findings provide a platform for interrogation of host and pathogen transcriptomes at the site of infection, allowing insight into mechanisms of inflammation in human disease.
]]></description>
<dc:creator>Montoya, D.</dc:creator>
<dc:creator>Andrade, P. R.</dc:creator>
<dc:creator>Silva, B. J. A.</dc:creator>
<dc:creator>Teles, R. M. B.</dc:creator>
<dc:creator>Bryson, B.</dc:creator>
<dc:creator>Sadanand, S.</dc:creator>
<dc:creator>Noel, T.</dc:creator>
<dc:creator>Lu, J.</dc:creator>
<dc:creator>Sarno, E.</dc:creator>
<dc:creator>Arnvig, K. B.</dc:creator>
<dc:creator>Young, D.</dc:creator>
<dc:creator>Lahiri, R.</dc:creator>
<dc:creator>Williams, D. L.</dc:creator>
<dc:creator>Fortune, S.</dc:creator>
<dc:creator>Bloom, B. R.</dc:creator>
<dc:creator>Pellegrini, M.</dc:creator>
<dc:creator>Modlin, R. L.</dc:creator>
<dc:date>2018-06-22</dc:date>
<dc:identifier>doi:10.1101/354407</dc:identifier>
<dc:title><![CDATA[Dual RNAseq of human leprosy lesions identifies bacterial determinants linked to host immune response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/353599v1?rss=1">
<title>
<![CDATA[
Transcriptome-wide profiling of mammalian spliceosome and branchpoints with iCLIP 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/353599v1?rss=1"
</link>
<description><![CDATA[
Studies of spliceosomal interactions are challenging due to their dynamic nature. Here we employed spliceosome iCLIP, which immunoprecipitates SmB along with snRNPs and auxiliary RNA binding proteins (RBPs), to map human spliceosome engagement with snRNAs and pre-mRNAs. This identified over 50,000 branchpoints (BPs) that have canonical sequence and structural features. Moreover, it revealed 7 binding peaks around BPs and splice sites, each precisely overlapping with binding profiles of specific splicing factors. We show how the binding patterns of these RBPs are affected by the position and strength of BPs. For example, strong or proximally located BPs preferentially bind SF3 rather than U2AF complex. Notably, these effects are partly neutralized during spliceosomal assembly in a way that depends on the core spliceosomal protein PRPF8. These insights exemplify spliceosome iCLIP as a broadly applicable method for transcriptomic studies of splicing mechanisms.
]]></description>
<dc:creator>Briese, M.</dc:creator>
<dc:creator>Haberman, N.</dc:creator>
<dc:creator>Sibley, C.</dc:creator>
<dc:creator>Chakrabarti, A.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Konig, J.</dc:creator>
<dc:creator>Perera, D.</dc:creator>
<dc:creator>Wickramasinghe, V.</dc:creator>
<dc:creator>Venkitaraman, A.</dc:creator>
<dc:creator>Luscombe, N.</dc:creator>
<dc:creator>Smith, C.</dc:creator>
<dc:creator>Curk, T.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:date>2018-06-22</dc:date>
<dc:identifier>doi:10.1101/353599</dc:identifier>
<dc:title><![CDATA[Transcriptome-wide profiling of mammalian spliceosome and branchpoints with iCLIP]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/360073v1?rss=1">
<title>
<![CDATA[
Nanoscale Polarization of the Vaccinia Virus Entry Fusion Complex Drives Efficient Fusion 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/360073v1?rss=1"
</link>
<description><![CDATA[
The spatial distribution of binding and fusion proteins on most viruses and the functional relevance of this organization remains largely unexplored. Employing super-resolution microscopy we define the nanoscale membrane architecture of the prototypic poxvirus, vaccinia. We show that binding and entry fusion complex (EFC) proteins are organized into distinct functional domains with fusion proteins polarized to the tips of virions. Repression of individual EFC components disrupted fusion protein polarization, correlating with a loss of fusion activity. Repression of vaccinia A27, a non-EFC protein implicated in fusion, revealed that disruption of EFC localization impacts virus fusion pore formation. We propose that the polarized distribution of EFCs is essential for poxvirus fusion efficiency.
]]></description>
<dc:creator>Gray, R.</dc:creator>
<dc:creator>Albrecht, D.</dc:creator>
<dc:creator>Beerli, C.</dc:creator>
<dc:creator>Cohen, G.</dc:creator>
<dc:creator>Henriques, R.</dc:creator>
<dc:creator>Mercer, J.</dc:creator>
<dc:date>2018-07-01</dc:date>
<dc:identifier>doi:10.1101/360073</dc:identifier>
<dc:title><![CDATA[Nanoscale Polarization of the Vaccinia Virus Entry Fusion Complex Drives Efficient Fusion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/367458v1?rss=1">
<title>
<![CDATA[
γ-proteobacteria eject their polar flagella under nutrient depletion, retaining flagellar motor relic structures 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/367458v1?rss=1"
</link>
<description><![CDATA[
Bacteria switch only intermittently to motile planktonic lifestyles under favourable conditions. Under chronic nutrient deprivation, however, bacteria orchestrate a switch to stationary phase, conserving energy by altering metabolism and stopping motility. About two-thirds of bacteria use flagella to swim, but how bacteria deactivate this large-molecular machine remains poorly studied. Here we describe the previously unreported ejection of polar sodium-driven motors by {gamma}-proteobacteria. We show that these bacteria eject their flagella at the base of the flagellar hook when nutrients are depleted, leaving a relic of a former flagellar motor in the outer membrane. Subtomogram averages of the full motor and relic reveal that this is an active process, as a plug protein appears in the relic, likely to prevent leakage across their outer membrane. We show that this is a widespread phenomenon demonstrated by the appearance of relic structures in varied {gamma}-proteobacteria including Plesiomonas shigelloides, Vibrio cholerae, Vibrio fischeri, Shewanella putrefaciens and Pseudomonas aeruginosa.
]]></description>
<dc:creator>Ferreira, J. L.</dc:creator>
<dc:creator>Gao, F. Z.</dc:creator>
<dc:creator>Rossmann, F. M.</dc:creator>
<dc:creator>Nans, A.</dc:creator>
<dc:creator>Brenzinger, S.</dc:creator>
<dc:creator>Hosseini, R.</dc:creator>
<dc:creator>Briegel, A.</dc:creator>
<dc:creator>Thormann, K. M.</dc:creator>
<dc:creator>Rosenthal, P. B.</dc:creator>
<dc:creator>Beeby, M.</dc:creator>
<dc:date>2018-07-18</dc:date>
<dc:identifier>doi:10.1101/367458</dc:identifier>
<dc:title><![CDATA[γ-proteobacteria eject their polar flagella under nutrient depletion, retaining flagellar motor relic structures]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/370999v1?rss=1">
<title>
<![CDATA[
Bcl6 promotes neurogenic conversion through transcriptional repression of multiple self-renewal-promoting extrinsic pathways. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/370999v1?rss=1"
</link>
<description><![CDATA[
During neurogenesis, progenitors switch from self-renewal to differentiation through the interplay of intrinsic and extrinsic cues, but how these are integrated remains poorly understood. Here we combine whole genome transcriptional and epigenetic analyses with in vivo functional studies and show that Bcl6, a transcriptional repressor known to promote neurogenesis, acts as a key driver of the neurogenic transition through direct silencing of a selective repertoire of genes belonging to multiple extrinsic pathways promoting self-renewal, most strikingly the Wnt pathway. At the molecular level, Bcl6 acts through both generic and pathway-specific mechanisms. Our data identify a molecular logic by which a single cell-intrinsic factor ensures robustness of neural cell fate transition by decreasing responsiveness to the extrinsic pathways that favor self-renewal.
]]></description>
<dc:creator>Bonnefont, J.</dc:creator>
<dc:creator>Tiberi, L.</dc:creator>
<dc:creator>van den Ameele, J.</dc:creator>
<dc:creator>Potier, D.</dc:creator>
<dc:creator>Gaber, Z.</dc:creator>
<dc:creator>Lin, X.</dc:creator>
<dc:creator>Bilheu, A.</dc:creator>
<dc:creator>Herpoel, A.</dc:creator>
<dc:creator>Velez Bravo, F.</dc:creator>
<dc:creator>Guillemot, F.</dc:creator>
<dc:creator>Aerts, S.</dc:creator>
<dc:creator>Vanderhaeghen, P.</dc:creator>
<dc:date>2018-07-18</dc:date>
<dc:identifier>doi:10.1101/370999</dc:identifier>
<dc:title><![CDATA[Bcl6 promotes neurogenic conversion through transcriptional repression of multiple self-renewal-promoting extrinsic pathways.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/370056v1?rss=1">
<title>
<![CDATA[
Drug susceptibility testing and mortality in patients treated for tuberculosis in high-burden countries 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/370056v1?rss=1"
</link>
<description><![CDATA[
BackgroundDrug resistance and HIV co-infection are challenges for the global control of tuberculosis.nnMethodsWe collected Mycobacterium tuberculosis isolates from adult patients in Cote dIvoire, Democratic Republic of the Congo, Kenya, Nigeria, South Africa, Peru, and Thailand, stratified by HIV status and tuberculosis drug resistance. Molecular or phenotypic drug susceptibility testing (DST) was done locally and at the Swiss tuberculosis reference laboratory. We examined mortality during treatment according to DST results and treatment adequacy in logistic regression models adjusting for sex, age, sputum microscopy and HIV status.nnFindings634 tuberculosis patients were included; median age was 33.2 years, 239 (37.7%) were female, 272 (42.9%) HIV-positive and 69 (10.9%) patients died. Based on the reference laboratory DST, 394 (62.2%) strains were pan-susceptible, 45 (7.1%) mono-resistant, 163 (25.7%) multidrug-resistant (MDR-TB), and 30 (4.7%) had pre-extensive or extensive drug resistance (pre-XDR/XDR-TB). Results of reference and local laboratories were discordant in 121 (19.1%) cases, corresponding to a sensitivity of 84.3% and a specificity of 90.8%. In patients with drug-resistant tuberculosis, discordant results were associated with increased mortality (risk ratio 1.81; 95% CI 1.07-3.07). In logistic regression, compared to adequately treated patients with pan-susceptible strains, the adjusted odds ratio for death was 4.23 (95% CI 2.16-8.29) for adequately treated patients with drug-resistant strains and 21.54 (95% CI 3.36-138.1) for inadequately treated patients with drug-resistant strains. HIV status was not associated with mortality.nnInterpretationUsing a reference laboratory standard, inaccurate DST leading to inappropriate treatment of drug-resistant tuberculosis, but not HIV infection, contributed to mortality.
]]></description>
<dc:creator>Zuercher, K.</dc:creator>
<dc:creator>Ballif, M.</dc:creator>
<dc:creator>Fenner, L.</dc:creator>
<dc:creator>Borrell, S.</dc:creator>
<dc:creator>Keller, P. M.</dc:creator>
<dc:creator>Gnokoro, J.</dc:creator>
<dc:creator>Marcy, O.</dc:creator>
<dc:creator>Yotebieng, M.</dc:creator>
<dc:creator>Diero, L.</dc:creator>
<dc:creator>Carter, E. J.</dc:creator>
<dc:creator>Rockwood, N.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Cox, H.</dc:creator>
<dc:creator>Ezati, N.</dc:creator>
<dc:creator>Abimiku, A. l. G.</dc:creator>
<dc:creator>Collantes, J.</dc:creator>
<dc:creator>Avihingsanon, A.</dc:creator>
<dc:creator>Kawkitinarong, K.</dc:creator>
<dc:creator>Reinhard, M.</dc:creator>
<dc:creator>Hoemke, R.</dc:creator>
<dc:creator>Huebner, R.</dc:creator>
<dc:creator>Gagneux, S.</dc:creator>
<dc:creator>Boettger, E. C.</dc:creator>
<dc:creator>Egger, M.</dc:creator>
<dc:creator>on behalf of the International Epidemiology Databases to Evaluate AIDS (IeDEA),</dc:creator>
<dc:date>2018-07-18</dc:date>
<dc:identifier>doi:10.1101/370056</dc:identifier>
<dc:title><![CDATA[Drug susceptibility testing and mortality in patients treated for tuberculosis in high-burden countries]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/374215v1?rss=1">
<title>
<![CDATA[
Antibiotic resistance evasion is explained by rare mutation frequency and not by lack of compensatory mechanisms 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/374215v1?rss=1"
</link>
<description><![CDATA[
Drug resistant infections represent one of the most challenging medical problems of our time. D-cycloserine is an antibiotic used for decades without appearance and dissemination of antibiotic resistant strains, making it an ideal model compound to understand what drives resistance evasion. We investigated why Mycobacterium tuberculosis fails to become resistant to D-cycloserine. To address this question we employed a combination of bacterial genetics, genomics, biochemistry and fitness analysis in vitro, in macrophages and in mice. Altogether, our results suggest that the ultra-low mutation frequency associated with D-cycloserine resistance is the dominant factor delaying the appearance of clinical resistance to this antibiotic in bacteria infecting humans, and not lack of potential compensatory mechanisms.nnOne Sentence SummaryWe show that the lack of D-cycloserine resistance in Mycobacterium tuberculosis is due its ultra-low mutation frequency rather than lack of compensatory mechanisms.
]]></description>
<dc:creator>Carvalho, L. P.</dc:creator>
<dc:creator>Evangelopoulos, D.</dc:creator>
<dc:creator>Prosser, G.</dc:creator>
<dc:creator>Rodgers, A.</dc:creator>
<dc:creator>Dagg, B.</dc:creator>
<dc:creator>Khatri, B.</dc:creator>
<dc:creator>Ho, M. M.</dc:creator>
<dc:creator>Gutierrez, M.</dc:creator>
<dc:creator>Cortes, T.</dc:creator>
<dc:date>2018-07-23</dc:date>
<dc:identifier>doi:10.1101/374215</dc:identifier>
<dc:title><![CDATA[Antibiotic resistance evasion is explained by rare mutation frequency and not by lack of compensatory mechanisms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/375147v1?rss=1">
<title>
<![CDATA[
Survival of the frequent at finite population size and mutation rate: filing the gap between quasispecies and monomorphic regimes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/375147v1?rss=1"
</link>
<description><![CDATA[
In recent years, there has been increased attention on the non-trivial role that genotype-phenotype maps play in the course of evolution, where natural selection acts on phenotypes, but variation arises at the level of mutations. Understanding such mappings is arguably the next missing piece in a fully predictive theory of evolution. Although there are theoretical descriptions of such mappings for the monomorphic (N << 1) and deterministic or very strong mutation (N [&gt;&gt;&gt;] 1) limit, given by developments of Iwasas free fitness and quasispecies theories, respectively, there is no general description for the intermediate regime where N ~ 1. In this paper, we address this by transforming Wrights well-known stationary distribution of genotypes under selection and mutation to give the probability distribution of phenotypes, assuming a general genotype-phenotype map. The resultant distribution shows that the degeneracies of each phenotype appear by weighting the mutation term; this gives rise to a bias towards phenotypes of larger degeneracy analogous to quasispecies theory, but at finite population size. On the other hand we show that as population size is decreased, again phenotypes of higher degeneracy are favoured, which is a finite mutation description of the effect of sequence entropy in the monomorphic limit. We also for the first time (to the authors knowledge) provide an explicit derivation of Wrights stationary distribution of the frequencies of multiple alleles.
]]></description>
<dc:creator>Khatri, B. S.</dc:creator>
<dc:date>2018-07-23</dc:date>
<dc:identifier>doi:10.1101/375147</dc:identifier>
<dc:title><![CDATA[Survival of the frequent at finite population size and mutation rate: filing the gap between quasispecies and monomorphic regimes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/375832v1?rss=1">
<title>
<![CDATA[
A Ubiquitin-Binding Domain that Does Not Bind Ubiquitin 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/375832v1?rss=1"
</link>
<description><![CDATA[
Ubiquitylation, the post-translational linkage of ubiquitin moieties to lysines in target proteins, helps regulate a myriad of biological processes. Ubiquitin, and sometimes ubiquitin-homology domains, are recognized by ubiquitin-binding domains, including CUE domains. CUE domains are thus generally thought to function exclusively by mediating interactions with ubiquitylated proteins. The chromatin remodeler, SMARCAD1, interacts with KAP1, a transcriptional corepressor. We show that the SMARCAD1-KAP1 interaction is direct and involves the first SMARCAD1 CUE domain (CUE1) and the RBCC domain of KAP1. A structural model of the minimal KAP1 RBCC-SMARCAD1 CUE1 complex based on X-ray crystallography analysis is presented. Remarkably, the CUE1 domain, which resembles a canonical CUE domain, recognizes 2 clusters of exposed hydrophobic residues on KAP1, but these are presented in the context of a coiled-coil domain, not in a structure resembling ubiquitin. Together, these data challenge the well-established dogma that CUE domains exclusively recognize the ubiquitin-fold.
]]></description>
<dc:creator>Lim, M.</dc:creator>
<dc:creator>Newman, J. A.</dc:creator>
<dc:creator>Williams, H. L.</dc:creator>
<dc:creator>Aitkenhead, H.</dc:creator>
<dc:creator>Gileadi, O.</dc:creator>
<dc:creator>Svejstrup, J.</dc:creator>
<dc:date>2018-07-24</dc:date>
<dc:identifier>doi:10.1101/375832</dc:identifier>
<dc:title><![CDATA[A Ubiquitin-Binding Domain that Does Not Bind Ubiquitin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/378133v1?rss=1">
<title>
<![CDATA[
Functional cross-talk between allosteric effects of activating and inhibiting ligands underlies PKM2 regulation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/378133v1?rss=1"
</link>
<description><![CDATA[
Allosteric regulation is central to the role of the glycolytic enzyme pyruvate kinase M2 (PKM2) in cellular metabolism. Multiple activating and inhibitory allosteric ligands regulate PKM2 activity by controlling the equilibrium between high activity tetramers and low activity dimers and monomers. However, it remains elusive how allosteric inputs upon simultaneous binding of different ligands are integrated to regulate PKM2 activity. Here, we show that, in the presence of the allosteric inhibitor L-phenylalanine (Phe), the activator fructose 1,6-bisphosphate (FBP) can induce PKM2 tetramerisation, but fails to maximally increase enzymatic activity. Guided by a new computational framework we developed to identify residues that mediate FBP-induced allostery, we generated two PKM2 mutants, A327S and C358A, in which activation by FBP remains intact but cannot be attenuated by Phe. Our findings demonstrate a role for residues involved in FBP-induced allostery in enabling the integration of allosteric input from Phe and reveal a mechanism that underlies the co-ordinate regulation of PKM2 activity by multiple allosteric ligands.
]]></description>
<dc:creator>Macpherson, J. A.</dc:creator>
<dc:creator>Theisen, A.</dc:creator>
<dc:creator>Masino, L.</dc:creator>
<dc:creator>Fets, L.</dc:creator>
<dc:creator>Driscoll, P. C.</dc:creator>
<dc:creator>Encheva, V.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Martin, S. R.</dc:creator>
<dc:creator>Kleinjung, J.</dc:creator>
<dc:creator>Barran, P. E.</dc:creator>
<dc:creator>Fraternali, F.</dc:creator>
<dc:creator>Anastasiou, D.</dc:creator>
<dc:date>2018-07-26</dc:date>
<dc:identifier>doi:10.1101/378133</dc:identifier>
<dc:title><![CDATA[Functional cross-talk between allosteric effects of activating and inhibiting ligands underlies PKM2 regulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/385278v1?rss=1">
<title>
<![CDATA[
Nitrogen utilisation by the metabolic generalist pathogen Mycobacterium tuberculosis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/385278v1?rss=1"
</link>
<description><![CDATA[
Bacterial metabolism is fundamental to pathogenesis and has a dominant effect on bacterial killing by antibiotics. Here we explore how Mycobacterium tuberculosis utilises amino acids as nitrogen sources, using a combination of bacterial physiology and stable isotope tracing coupled to liquid chromatography mass spectrometry metabolomics methods. Our results define core properties of the nitrogen metabolic network from M. tuberculosis, such as: (i) the lack of homeostatic control of amino acid pool sizes; (ii) similar rates of utilisation of different amino acids as sole nitrogen sources; (iii) improved nitrogen utilisation from amino acids compared to ammonium; and (iv) co-metabolism of nitrogen sources. Finally, we discover that alanine dehydrogenase, is involved in ammonium assimilation in M. tuberculosis, in addition to its essential role in alanine utilisation. This study represents the first in-depth analysis of nitrogen source utilisation by metabolic generatlist M. tuberculosis and reveals a flexible metabolic network with characteristics that are likely product of evolution in the human host.
]]></description>
<dc:creator>Agapova, A.</dc:creator>
<dc:creator>Hunt, D. M.</dc:creator>
<dc:creator>Petridis, M.</dc:creator>
<dc:creator>Garza-Garcia, A.</dc:creator>
<dc:creator>Sohaskey, C. D.</dc:creator>
<dc:creator>Carvalho, L. P.</dc:creator>
<dc:date>2018-08-05</dc:date>
<dc:identifier>doi:10.1101/385278</dc:identifier>
<dc:title><![CDATA[Nitrogen utilisation by the metabolic generalist pathogen Mycobacterium tuberculosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/385344v1?rss=1">
<title>
<![CDATA[
Mapping the human kinome in response to DNA damage 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/385344v1?rss=1"
</link>
<description><![CDATA[
We provide a catalog for the effects of the human kinome on cell survival in response to DNA damaging agents, selected to cover all major DNA repair pathways. By treating 313 kinase-deficient cell lines with ten diverse DNA damaging agents, including seven commonly used chemotherapeutics, we were able to identify kinase specific vulnerabilities and resistances. In order to identify novel synthetic lethal interactions, we investigate the cellular response to carmustine for 25 cell lines, by establishing a phenotypic FACS assay designed to mechanistically investigate and validate gene-drug interactions. We show apoptosis, cell cycle, DNA damage and proliferation after alkylation or crosslink-induced damage for selected cell lines and rescue the cellular sensitivity of DYRK4, EPHB6, MARK3, PNCK as a proof of principle for our study. Our data suggest that some cancers with inactivated DYRK4, EPHB6, MARK3 or PNCK gene could be particularly vulnerable to treatment by alkylating chemotherapeutic agents carmustine or temozolomide.
]]></description>
<dc:creator>Owusu, M.</dc:creator>
<dc:creator>Bannauer, P.</dc:creator>
<dc:creator>Mourikis, A.</dc:creator>
<dc:creator>Jones, A.</dc:creator>
<dc:creator>Ferreira da Silva, J.</dc:creator>
<dc:creator>Caldera, M.</dc:creator>
<dc:creator>Wiedner, M.</dc:creator>
<dc:creator>Lardeau, C.-H.</dc:creator>
<dc:creator>Menche, J.</dc:creator>
<dc:creator>Kubicek, S.</dc:creator>
<dc:creator>Ciccarelli, F.</dc:creator>
<dc:creator>Loizou, J.</dc:creator>
<dc:date>2018-08-06</dc:date>
<dc:identifier>doi:10.1101/385344</dc:identifier>
<dc:title><![CDATA[Mapping the human kinome in response to DNA damage]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/387027v1?rss=1">
<title>
<![CDATA[
Target-specific precision of CRISPR-mediated genome editing 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/387027v1?rss=1"
</link>
<description><![CDATA[
The CRISPR-Cas9 system has successfully been adapted to edit the genome of various organisms. However, our ability to predict editing accuracy, efficacy and outcome at specific sites is limited by an incomplete understanding of how the bacterial system interacts with eukaryotic genomes and DNA repair machineries. Here, we performed the largest comparison of indel profiles to date, examining over one thousand sites in the genome of human cells, and uncovered general principles guiding CRISPR-mediated DNA editing. We find that precision of DNA editing varies considerably among sites, with some targets showing one highly-preferred indel and others displaying a wide range of infrequent indels. Editing precision correlates with editing efficiency, homology-associated end-joining for both insertions and deletions, and a preference for single-nucleotide insertions. Precise targets and the identity of their preferred indel can be predicted based on simple rules that mainly depend on the fourth nucleotide upstream of the PAM sequence. Regardless of precision, site-specific indel profiles are highly robust and depend on both DNA sequence and chromatin features. Our findings have important implications for clinical applications of CRISPR technology and reveal general patterns of broken end-joining that can inform us on DNA repair mechanisms in human cells.
]]></description>
<dc:creator>Chakrabarti, A. M.</dc:creator>
<dc:creator>Henser-Brownhill, T.</dc:creator>
<dc:creator>Monserrat, J.</dc:creator>
<dc:creator>Poetsch, A. R.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:creator>Scaffidi, P.</dc:creator>
<dc:date>2018-08-09</dc:date>
<dc:identifier>doi:10.1101/387027</dc:identifier>
<dc:title><![CDATA[Target-specific precision of CRISPR-mediated genome editing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/389858v1?rss=1">
<title>
<![CDATA[
The Network of Cancer Genes (NCG): a comprehensive catalogue of known and candidate cancer genes from cancer sequencing screens. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/389858v1?rss=1"
</link>
<description><![CDATA[
The Network of Cancer Genes (NCG) is a manually curated repository of 2,372 genes whose somatic modifications have a known or predicted cancer driver role. These genes were collected from 275 publications, including two sources of known cancer genes and 273 cancer sequencing screens of 119 cancer types in 31 primary sites from 34,905 cancer donors. This represents a more than 1.5-fold increase in content as compared to the previous version. NCG also annotates properties of cancer genes, such as duplicability, evolutionary origin, RNA and protein expression, miRNA and protein interactions, protein function and essentiality. NCG is accessible at http://ncg.kcl.ac.uk/.
]]></description>
<dc:creator>Ciccarelli, F. D.</dc:creator>
<dc:creator>Kuppili Venkata, S.</dc:creator>
<dc:creator>Repana, D.</dc:creator>
<dc:creator>Nulsen, J.</dc:creator>
<dc:creator>Dressler, L.</dc:creator>
<dc:creator>Bortolomeazzi, M.</dc:creator>
<dc:creator>Tourna, A.</dc:creator>
<dc:creator>Yakovleva, A.</dc:creator>
<dc:creator>Palmieri, T.</dc:creator>
<dc:date>2018-08-11</dc:date>
<dc:identifier>doi:10.1101/389858</dc:identifier>
<dc:title><![CDATA[The Network of Cancer Genes (NCG): a comprehensive catalogue of known and candidate cancer genes from cancer sequencing screens.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/394239v1?rss=1">
<title>
<![CDATA[
P-TEFb activation by RBM7 shapes a pro-survival transcriptional response to genotoxic stress 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/394239v1?rss=1"
</link>
<description><![CDATA[
Cellular DNA damage response (DDR) involves dramatic transcriptional alterations, the mechanisms of which remain ill-defined. Given the centrality of RNA polymerase II (Pol II) promoter-proximal pause release in transcriptional control, we evaluated its importance in DDR. Here we show that following genotoxic stress, the RNA-binding motif protein 7 (RBM7) stimulates Pol II elongation and promotes cell viability by activating the positive transcription elongation factor b (P-TEFb). This is mediated by genotoxic stress-enhanced binding of RBM7 to 7SK snRNA (7SK), the scaffold of the 7SK small nuclear ribonucleoprotein (7SK snRNP) which inhibits P-TEFb. In turn, P-TEFb relocates from 7SK snRNP to chromatin to induce transcription of short units including key DDR genes and multiple classes of non-coding RNAs. Critically, interfering with RBM7 or P-TEFb provokes cellular hypersensitivity to DNA damage-inducing agents through activation of apoptotic program. By alleviating the inhibition of P-TEFb, RBM7 thus facilitates Pol II elongation to enable a pro-survival transcriptional response that is crucial for cell fate upon genotoxic insult. Our work uncovers a new paradigm in stress-dependent control of Pol II pause release, and offers the promise for designing novel anti-cancer interventions using RBM7 and P-TEFb antagonists in combination with DNA-damaging chemotherapeutics.
]]></description>
<dc:creator>Bugai, A.</dc:creator>
<dc:creator>Quaresma, A. J.</dc:creator>
<dc:creator>Friedel, C. C.</dc:creator>
<dc:creator>Lenasi, V.</dc:creator>
<dc:creator>Sibley, C. R.</dc:creator>
<dc:creator>Kukanja, P.</dc:creator>
<dc:creator>Fujinaga, K.</dc:creator>
<dc:creator>Blasius, M.</dc:creator>
<dc:creator>Hennig, T.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Doelken, L.</dc:creator>
<dc:creator>Barboric, M.</dc:creator>
<dc:date>2018-08-17</dc:date>
<dc:identifier>doi:10.1101/394239</dc:identifier>
<dc:title><![CDATA[P-TEFb activation by RBM7 shapes a pro-survival transcriptional response to genotoxic stress]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/394692v1?rss=1">
<title>
<![CDATA[
Genome-Wide Reconstitution of Chromatin Transactions: RSC Preferentially disrupts H2A.Z-Containing Nucleosomes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/394692v1?rss=1"
</link>
<description><![CDATA[
Chromatin transactions are typically studied in vivo, or in vitro using artificial chromatin lacking the epigenetic complexity of the natural material. Attempting to bridge the gap between these approaches, we established a system for isolating the yeast genome as a library of mono-nucleosomes harboring the natural epigenetic signature, suitable for biochemical manipulation. Combined with deep sequencing, this library was used to investigate the intrinsic stability of individual nucleosomes, and - as proof of principle - the nucleosome preference of the chromatin remodeling complex, RSC. Our data indicate that the natural stability of nucleosomes differs greatly, with nucleosomes on tRNA genes and on promoters of protein-coding genes standing out as intrinsically unstable. Interestingly, RSC shows a distinct preference for nucleosomes derived from regions with a high density of histone variant H2A.Z, and this preference is indeed markedly diminished using nucleosomes from cells lacking H2A.Z. Importantly, the preference for H2A.Z remodeling/nucleosome ejection can also be reconstituted with recombinant nucleosome arrays. Together, our data indicate that, despite being separated from their genomic context, individual nucleosomes can retain their original identity as promoter- or TSS-nucleosomes. Besides shedding new light on nucleosome stability and the chromatin remodeler RSC, the simple experimental system outlined here should be generally applicable to the study of chromatin transactions.
]]></description>
<dc:creator>Cakiroglu, A.</dc:creator>
<dc:creator>Clapier, C.</dc:creator>
<dc:creator>Ehrensberger, A.</dc:creator>
<dc:creator>Darbo, E.</dc:creator>
<dc:creator>Cairns, B.</dc:creator>
<dc:creator>Luscombe, N.</dc:creator>
<dc:creator>Svejstrup, J.</dc:creator>
<dc:date>2018-08-17</dc:date>
<dc:identifier>doi:10.1101/394692</dc:identifier>
<dc:title><![CDATA[Genome-Wide Reconstitution of Chromatin Transactions: RSC Preferentially disrupts H2A.Z-Containing Nucleosomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/397000v1?rss=1">
<title>
<![CDATA[
CryoEM structures of open dimers of Gyrase A in complex with DNA illuminate mechanism of strand passage 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/397000v1?rss=1"
</link>
<description><![CDATA[
Gyrase is a unique type IIA topoisomerase that uses ATP hydrolysis to maintain the negatively supercoiled state of bacterial DNA. In order to perform its function, gyrase undergoes a sequence of conformational changes that consist of concerted gate openings, DNA cleavage, and DNA strand passage events. Structures where the transported DNA molecule (T-segment) is trapped by the A subunit have not been observed. Here we present the cryoEM structures of two oligomeric complexes of open gyrase A dimers and DNA. The protein subunits in these complexes were solved to 4 [A] and 5.16 [A] resolution. One of the complexes traps a linear DNA molecule, a putative T-segment, which interacts with the open gyrase A dimers in two states, representing steps either prior to or after passage through the DNA-gate. The structures locate the T-segment in important intermediate conformations of the catalytic cycle and provide insights into gyrase-DNA interactions and mechanism.
]]></description>
<dc:creator>Soczek, K. M.</dc:creator>
<dc:creator>Grant, T.</dc:creator>
<dc:creator>Rosenthal, P. B.</dc:creator>
<dc:creator>Mondragon, A.</dc:creator>
<dc:date>2018-08-21</dc:date>
<dc:identifier>doi:10.1101/397000</dc:identifier>
<dc:title><![CDATA[CryoEM structures of open dimers of Gyrase A in complex with DNA illuminate mechanism of strand passage]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/397489v1?rss=1">
<title>
<![CDATA[
A divergent kinase lacking the glycine-rich loop regulates membrane ultrastructure of the Toxoplasma parasitophorous vacuole 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/397489v1?rss=1"
</link>
<description><![CDATA[
Apicomplexan parasites replicate within a protective organelle called the parasitophorous vacuole (PV). The Toxoplasma gondii PV is filled with a network of tubulated membranes, which are thought to facilitate trafficking of effectors and nutrients. Despite being critical to parasite virulence, there is scant mechanistic understanding of the networks functions. Here, we identify the parasite secreted kinase WNG1 as a critical regulator of tubular membrane biogenesis. WNG1 family members adopt an atypical protein kinase fold lacking the glycine rich ATP-binding loop that is required for catalysis in canonical kinases. Unexpectedly, we find that WNG1 is an active protein kinase that localizes to the PV lumen and phosphorylates PV-resident proteins, several of which are essential for the formation of a functional intravacuolar network. Moreover, we show that WNG1-dependent phosphorylation of these proteins is required for their membrane association, and thus their ability to tubulate membranes. Consequently, WNG1 knockout parasites have an aberrant PV membrane ultrastructure. Collectively, our results describe a unique family of Toxoplasma kinases and implicate phosphorylation of secreted proteins as a mechanism of regulating PV formation during parasite infection.
]]></description>
<dc:creator>Beraki, T.</dc:creator>
<dc:creator>Hu, X.</dc:creator>
<dc:creator>Broncel, M.</dc:creator>
<dc:creator>Young, J. C.</dc:creator>
<dc:creator>O'Shaughnessy, W. J.</dc:creator>
<dc:creator>Borek, D. M.</dc:creator>
<dc:creator>Treeck, M.</dc:creator>
<dc:creator>Reese, M. L.</dc:creator>
<dc:date>2018-08-22</dc:date>
<dc:identifier>doi:10.1101/397489</dc:identifier>
<dc:title><![CDATA[A divergent kinase lacking the glycine-rich loop regulates membrane ultrastructure of the Toxoplasma parasitophorous vacuole]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/399865v1?rss=1">
<title>
<![CDATA[
EGFR SIGNALING COORDINATES PATTERNING WITH CELL SURVIVAL DURING DROSOPHILA EPIDERMAL DEVELOPMENT 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/399865v1?rss=1"
</link>
<description><![CDATA[
Extensive apoptosis is often seen in patterning mutants, suggesting that tissues can detect and eliminate potentially harmful mis-specified cells. Here we show that the pattern of apoptosis in the embryonic epidermis of Drosophila is not a response to fate mis-specification but can instead be explained by the limiting availability of pro-survival signalling molecules released from locations determined by patterning information. In wild type embryos, the segmentation cascade elicits the segmental production of several EGFR ligands, including the TGF-alpha, Spitz and the Neuregulin, Vein. This leads to an undulating pattern of signalling activity, which prevents expression of the pro-apoptotic gene hid throughout the epidermis. In segmentation mutants, where specific peaks of EGFR ligands fail to form, gaps in signalling activity appear, leading to coincident hid upregulation and subsequent cell death. These data provide a mechanistic understanding of how cell survival, and thus appropriate tissue size, is made contingent on correct patterning.
]]></description>
<dc:creator>Crossman, S. H.</dc:creator>
<dc:creator>Streichan, S.</dc:creator>
<dc:creator>vincent, J.-P.</dc:creator>
<dc:date>2018-08-24</dc:date>
<dc:identifier>doi:10.1101/399865</dc:identifier>
<dc:title><![CDATA[EGFR SIGNALING COORDINATES PATTERNING WITH CELL SURVIVAL DURING DROSOPHILA EPIDERMAL DEVELOPMENT]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/408450v1?rss=1">
<title>
<![CDATA[
An Artificial Intelligence Workflow for Defining Host-Pathogen Interactions 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/408450v1?rss=1"
</link>
<description><![CDATA[
For image-based infection biology, accurate unbiased quantification of host-pathogen interactions is essential, yet often performed manually or using limited enumeration employing simple image analysis algorithms based on image segmentation. Host protein recruitment to pathogens is often refractory to accurate automated assessment due to its heterogeneous nature. An intuitive intelligent image analysis program to assess host protein recruitment within general cellular pathogen defense is lacking. We present HRMAn (Host Response to Microbe Analysis), an open-source image analysis platform based on machine learning algorithms and deep learning. We show that HRMAn has the capability to learn phenotypes from the data, without relying on researcher-based assumptions. Using Toxoplasma gondii and Salmonella typhimurium we demonstrate HRMAns capacity to recognize, classify and quantify pathogen killing, replication and cellular defense responses.
]]></description>
<dc:creator>Fisch, D. H.</dc:creator>
<dc:creator>Yakimovich, A.</dc:creator>
<dc:creator>Clough, B.</dc:creator>
<dc:creator>Wright, J.</dc:creator>
<dc:creator>Bunyan, M.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>Mercer, J.</dc:creator>
<dc:creator>Frickel, E.-M.</dc:creator>
<dc:date>2018-09-05</dc:date>
<dc:identifier>doi:10.1101/408450</dc:identifier>
<dc:title><![CDATA[An Artificial Intelligence Workflow for Defining Host-Pathogen Interactions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/404798v1?rss=1">
<title>
<![CDATA[
Function and essentiality of Plasmodium falciparum Plasmepsin V 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/404798v1?rss=1"
</link>
<description><![CDATA[
The malaria parasite replicates within erythrocytes. The pathogenesis of clinical malaria is in large part due to the capacity of the parasite to remodel its host cell. To do this, intraerythrocytic stages of Plasmodium falciparum export more than 300 proteins that dramatically alter the morphology of the infected erythrocyte as well as its mechanical and adhesive properties. P. falciparum plasmepsin V (PfPMV) is an aspartic protease that processes proteins for export into the host erythrocyte and is thought to play a key role in parasite virulence and survival. However, although standard techniques for gene disruption as well as conditional protein knockdown have been previously attempted with the pfpmv gene, complete gene removal or knockdown was not achieved so direct genetic proof that PMV is an essential protein has not yet been established. Here we have used a conditional gene excision approach combining CRISPR-Cas9 gene editing and DiCre-mediated recombination to functionally inactivate the pfpmv gene. The resulting mutant parasites displayed a severe growth defect. Detailed phenotypic analysis showed that development of the mutant parasites was arrested at the ring-to-trophozoite transition in the erythrocytic cycle following gene excision, likely due to a defect in protein export. Our findings are the first to elucidate the effects of PMV gene disruption, showing that it is essential for parasite viability in asexual blood stages. The mutant parasites can now be used as a platform to further dissect the Plasmodium protein export pathway.
]]></description>
<dc:creator>Boonyalai, N.</dc:creator>
<dc:creator>Collins, C. R.</dc:creator>
<dc:creator>Hackett, F.</dc:creator>
<dc:creator>Withers-Martinez, C.</dc:creator>
<dc:creator>Blackman, M. J.</dc:creator>
<dc:date>2018-09-06</dc:date>
<dc:identifier>doi:10.1101/404798</dc:identifier>
<dc:title><![CDATA[Function and essentiality of Plasmodium falciparum Plasmepsin V]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/419630v1?rss=1">
<title>
<![CDATA[
Apical caspase reporters uncover unknown stages of apoptosis and enable ready visualization of undead cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/419630v1?rss=1"
</link>
<description><![CDATA[
The caspase-mediated regulation of many cellular processes, including apoptosis, justifies the substantial interest in understanding all of the biological features of these enzymes. To complement functional assays, it is critical to identify caspase-activating cells in live tissues. Our work describes new caspase-reporters that, for the first time, provide direct information concerning the initial steps of the caspase activation cascade in Drosophila tissues. One of our caspase-sensors has capitalized on the rapid subcellular localization change of a fluorescent marker to uncover novel cellular apoptotic events. These refer to the actin-mediated positioning of the nucleus before cell delamination. The other construct has benefited from a caspase-induced nuclear translocation of a QF transcription factor. This feature enables the genetic manipulation of caspase-activating cells, whilst showing the spatio-temporal patterns of apical caspase activation. Collectively, our sensors offer new experimental opportunities that are already illuminating unknown aspects of caspase-dependent processes in apoptotic and non-apoptotic cellular scenarios.nnSummary statementWe describe a novel set of caspase sensors that directly detect early caspase activation. The exclusive features of our reporters uncovered unknown stages of apoptosis and properties of caspase-activating cells.
]]></description>
<dc:creator>Baena-Lopez, L. A.</dc:creator>
<dc:creator>Arthurton, L.</dc:creator>
<dc:creator>Bischoff, M.</dc:creator>
<dc:creator>Vincent, J.-P.</dc:creator>
<dc:creator>Alexandre, C.</dc:creator>
<dc:creator>McGregor, R.</dc:creator>
<dc:date>2018-09-17</dc:date>
<dc:identifier>doi:10.1101/419630</dc:identifier>
<dc:title><![CDATA[Apical caspase reporters uncover unknown stages of apoptosis and enable ready visualization of undead cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/426015v1?rss=1">
<title>
<![CDATA[
Id4 eliminates the pro-activation factor Ascl1 to maintain quiescence of adult hippocampal stem cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/426015v1?rss=1"
</link>
<description><![CDATA[
Quiescence is essential for the long-term maintenance of adult stem cells and tissue homeostasis. However, how stem cells maintain quiescence is still poorly understood. Here we show that stem cells in the dentate gyrus of the adult hippocampus actively transcribe the pro-activation factor Ascl1 regardless of their activation state. We found that the inhibitor of DNA binding protein Id4 suppresses Ascl1 activity in neural stem cell cultures. Id4 sequesters Ascl1 heterodimerisation partner E47, promoting Ascl1 protein degradation and neural stem cell quiescence. Accordingly, elimination of Id4 from stem cells in the adult hippocampus results in abnormal accumulation of Ascl1 protein and premature stem cell activation. We also found that multiple signalling pathways converge on the regulation of Id4 to reduce the activity of hippocampal stem cells. Id4 therefore maintains quiescence of adult neural stem cells, in sharp contrast with its role of promoting the proliferation of embryonic neural progenitors.
]]></description>
<dc:creator>Blomfield, I. M.</dc:creator>
<dc:creator>Rocamonde, B.</dc:creator>
<dc:creator>Masdeu, M. D. M.</dc:creator>
<dc:creator>Mulugeta, E.</dc:creator>
<dc:creator>Vaga, S.</dc:creator>
<dc:creator>van den Berg, D. L. C.</dc:creator>
<dc:creator>Huillard, E.</dc:creator>
<dc:creator>Guillemot, F.</dc:creator>
<dc:creator>Urban, N.</dc:creator>
<dc:date>2018-09-25</dc:date>
<dc:identifier>doi:10.1101/426015</dc:identifier>
<dc:title><![CDATA[Id4 eliminates the pro-activation factor Ascl1 to maintain quiescence of adult hippocampal stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/431684v1?rss=1">
<title>
<![CDATA[
Temporal control of cortico-thalamic neuron specification by regulation of Neurogenin activity and Polycomb repressive complexes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/431684v1?rss=1"
</link>
<description><![CDATA[
Neural progenitor cells (NPCs) in the embryonic mammalian neocortex generate different neuronal subtypes sequentially. A long-standing hypothesis to account for this temporal fate specification process is that NPCs change their differentiation potential over time. However, the molecular mechanisms underlying these temporal changes in NPC properties are poorly understood. Here we show that Neurogenin1 and Neurogenin2 (Neurog1/2), two proneural transcription factors expressed in NPCs throughout cortical neurogenesis, specify the identity of one of the first cortical neuron subtypes generated, layer 6 cortico-thalamic neurons (CTNs). We found that Neurog1/2 specify the CTN fate through regulation of the cortical fate determinants Fezf2 and Foxp2 and that this Neurog-induced programme becomes inactive after the period of CTN production. Two independent mechanisms contribute to the arrest of CTN neuron generation at the end of layer 6 neurogenesis, including a reduction in the transcriptional activity of Neurog1/2 and the deposition of epigenetic repressive modifications mediated by Polycomb repressive complexes at the Foxp2 gene. Therefore, the duration of production of a cortical neuron subtype is controlled by multiple locking mechanisms involving both transcriptional and epigenetic processes.
]]></description>
<dc:creator>Oishi, K.</dc:creator>
<dc:creator>van den Berg, D. L. C.</dc:creator>
<dc:creator>Guillemot, F.</dc:creator>
<dc:date>2018-10-01</dc:date>
<dc:identifier>doi:10.1101/431684</dc:identifier>
<dc:title><![CDATA[Temporal control of cortico-thalamic neuron specification by regulation of Neurogenin activity and Polycomb repressive complexes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/432674v1?rss=1">
<title>
<![CDATA[
NanoJ: a high-performance open-source super-resolution microscopy toolbox 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/432674v1?rss=1"
</link>
<description><![CDATA[
Super-resolution microscopy has become essential for the study of nanoscale biological processes. This type of imaging often requires the use of specialised image analysis tools to process a large volume of recorded data and extract quantitative information. In recent years, our team has built an open-source image analysis framework for super-resolution microscopy designed to combine high performance and ease of use. We named it NanoJ - a reference to the popular ImageJ software it was de-veloped for. In this paper, we highlight the current capabilities of NanoJ for several essential processing steps: spatio-temporal alignment of raw data (NanoJ-Core), super-resolution image re-construction (NanoJ-SRRF), image quality assessment (NanoJ-SQUIRREL), structural modelling (NanoJ-VirusMapper) and control of the sample environment (NanoJ-Fluidics). We expect to expand NanoJ in the future through the development of new tools designed to improve quantitative data analysis and measure the reliability of fluorescent microscopy studies.
]]></description>
<dc:creator>Laine, R.</dc:creator>
<dc:creator>Tosheva, K.</dc:creator>
<dc:creator>Gustafsson, N.</dc:creator>
<dc:creator>Gray, R. D. M.</dc:creator>
<dc:creator>Almada, P.</dc:creator>
<dc:creator>Albrecht, D.</dc:creator>
<dc:creator>Risa, G. T.</dc:creator>
<dc:creator>Hurtig, F.</dc:creator>
<dc:creator>Lindas, A.-C.</dc:creator>
<dc:creator>Baum, B.</dc:creator>
<dc:creator>Mercer, J.</dc:creator>
<dc:creator>Leterrier, C.</dc:creator>
<dc:creator>Pereira, P. M.</dc:creator>
<dc:creator>Culley, S.</dc:creator>
<dc:creator>Henriques, R.</dc:creator>
<dc:date>2018-10-01</dc:date>
<dc:identifier>doi:10.1101/432674</dc:identifier>
<dc:title><![CDATA[NanoJ: a high-performance open-source super-resolution microscopy toolbox]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/445411v1?rss=1">
<title>
<![CDATA[
Mitochondria form cholesterol-rich contact sites with the nucleus during retrograde response 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/445411v1?rss=1"
</link>
<description><![CDATA[
Mitochondria drive cellular adaptation to stress by retro-communicating with the nucleus. This process is known as Mitochondrial Retrograde Response (MRR) and is induced by mitochondrial dysfunctions which perturb cell signalling. MRR results in the nuclear stabilization and activation of pro-survival transcription factors such as the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB). Here we demonstrate that MRR is facilitated by the formation of contact sites between mitochondria and the nucleus which establish microdomains of communication between the two organelles. The 18kD Translocator Protein (TSPO), which de-ubiquitylates and stabilizes the mitochondrial network preventing its mitophagy-mediated segregation, is required for this interaction. The tethering TSPO enacts is mediated by the complex formed with the Protein Kinase A via the A-kinase anchoring protein Acyl-CoA Binding Domain Containing 3 (ACBD3) and allows the redistribution of cholesterol which sustains the pro-survival response by blocking NF-kB de-acetylation. This work proposes a new paradigm in the mitochondrial retro-communication by revealing the existence of contact sites between mitochondrial and the nucleus and a signalling role for cholesterol.
]]></description>
<dc:creator>Desai, R.</dc:creator>
<dc:creator>East, D. A.</dc:creator>
<dc:creator>Hardy, L.</dc:creator>
<dc:creator>Crosby, J.</dc:creator>
<dc:creator>Faccenda, D.</dc:creator>
<dc:creator>Alvarex, M. S.</dc:creator>
<dc:creator>Mainenti, M.</dc:creator>
<dc:creator>Hussey, L. K.</dc:creator>
<dc:creator>Bentham, R.</dc:creator>
<dc:creator>Szabadkai, G.</dc:creator>
<dc:creator>Zappulli, V.</dc:creator>
<dc:creator>Dhoot, G.</dc:creator>
<dc:creator>Fleck, R. A.</dc:creator>
<dc:creator>Vizcay-Barrena, G.</dc:creator>
<dc:creator>Smith, K.</dc:creator>
<dc:creator>Campanella, M.</dc:creator>
<dc:date>2018-10-17</dc:date>
<dc:identifier>doi:10.1101/445411</dc:identifier>
<dc:title><![CDATA[Mitochondria form cholesterol-rich contact sites with the nucleus during retrograde response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/450635v1?rss=1">
<title>
<![CDATA[
Fix your membrane receptor imaging: Actin cytoskeleton and CD4 membrane organization disruption by chemical fixation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/450635v1?rss=1"
</link>
<description><![CDATA[
Single-molecule localization microscopy (SMLM) techniques allow near molecular scale resolution (~ 20nm) as well as precise and robust analysis of protein organization at different scales. SMLM hardware, analytics and probes have been the focus of a variety of studies and are now commonly used in laboratories across the world. Protocol reliability and artefact identification are increasingly seen as important aspects of super-resolution microscopy. The reliability of these approaches thus requires in-depth evaluation so that biological findings are based on solid foundations. Here we explore how different fixation approaches that disrupt or preserve the actin cytoskeleton affect membrane protein organization. Using CD4 as a model, we show that fixation-mediated disruption of the actin cytoskeleton correlates with changes in CD4 membrane organization. We highlight how these artefacts are easy to overlook and how careful sample preparation is essential for extracting meaningful results from super-resolution microscopy.
]]></description>
<dc:creator>Pereira, P. M.</dc:creator>
<dc:creator>Albrecht, D.</dc:creator>
<dc:creator>Jacobs, C.</dc:creator>
<dc:creator>Marsh, M.</dc:creator>
<dc:creator>Mercer, J.</dc:creator>
<dc:creator>Henriques, R.</dc:creator>
<dc:date>2018-10-23</dc:date>
<dc:identifier>doi:10.1101/450635</dc:identifier>
<dc:title><![CDATA[Fix your membrane receptor imaging: Actin cytoskeleton and CD4 membrane organization disruption by chemical fixation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/472415v1?rss=1">
<title>
<![CDATA[
Single cell transcriptomics reveals spatial and temporal dynamics of gene expression in the developing mouse spinal cord 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/472415v1?rss=1"
</link>
<description><![CDATA[
The coordinated spatial and temporal regulation of gene expression in the vertebrate neural tube determines the identity of neural progenitors and the function and physiology of the neurons they generate. Progress has been made deciphering the gene regulatory programmes responsible for this process, however, the complexity of the tissue has hampered the systematic analysis of the network and the underlying mechanisms. To address this, we used single cell mRNA sequencing to profile cervical and thoracic regions of the developing mouse neural tube between embryonic days (e)9.5-e13.5. We confirmed the data accurately recapitulates neural tube development, allowing us to identify new markers for specific progenitor and neuronal populations. In addition, the analysis highlighted a previously underappreciated temporal component to the mechanisms generating neuronal diversity and revealed common features in the sequence of transcriptional events that lead to the differentiation of specific neuronal subtypes. Together the data provide a compendium of gene expression for classifying spinal cord cell types that will support future studies of neural tube development, function, and disease.
]]></description>
<dc:creator>Delile, J.</dc:creator>
<dc:creator>Rayon, T.</dc:creator>
<dc:creator>Melchionda, M.</dc:creator>
<dc:creator>Edwards, A.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Sagner, A.</dc:creator>
<dc:date>2018-11-16</dc:date>
<dc:identifier>doi:10.1101/472415</dc:identifier>
<dc:title><![CDATA[Single cell transcriptomics reveals spatial and temporal dynamics of gene expression in the developing mouse spinal cord]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/472266v1?rss=1">
<title>
<![CDATA[
Robust estimation of recent effective population size from number of independent origins in soft sweeps 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/472266v1?rss=1"
</link>
<description><![CDATA[
Estimating recent effective population size is of great importance in characterising and predicting the evolution of natural populations. Methods based on nucleotide diversity may underestimate current day effective population sizes due to historical bottlenecks, whilst methods that reconstruct demographic history typically only detect long-term variations. However, soft selective sweeps, which leave a fingerprint of mutational history by recurrent mutations on independent haplotype backgrounds, holds promise of an estimate more representative of recent population history. Here we present a simple and robust method of estimation based only on knowledge of the number of independent recurrent origins and the current frequency of the beneficial allele in a population sample, independent of the strength of selection and age of the mutation. Using a forward time theoretical framework, we show the mean number of origins is a function of{theta} = 2N and current allele frequency, through a simple equation, and the distribution is approximately Poisson. This estimate is robust to whether mutants pre-existed before selection arose, and is equally accurate for diploid populations with incomplete dominance. For fast (e.g., seasonal) demographic changes compared to time scale for fixation of the mutant allele, and for moderate peak-to-trough ratios, we show our constant population size estimate can be used to bound the maximum and minimum population size. Applied to the Vgsc gene of Anopheles gambiae, we estimate an effective population size of roughly 6 x 107, and including seasonal demographic oscillations, a minimum effective population size greater than 6 x 106 and a maximum less than 3 x 109.
]]></description>
<dc:creator>Khatri, B. S.</dc:creator>
<dc:creator>Burt, A.</dc:creator>
<dc:date>2018-11-19</dc:date>
<dc:identifier>doi:10.1101/472266</dc:identifier>
<dc:title><![CDATA[Robust estimation of recent effective population size from number of independent origins in soft sweeps]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/488056v1?rss=1">
<title>
<![CDATA[
The Spatio-Temporal Control of Zygotic Genome Activation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/488056v1?rss=1"
</link>
<description><![CDATA[
One of the earliest and most significant events in embryonic development is zygotic genome activation (ZGA). In several species, bulk transcription begins at the mid-blastula transition (MBT) when, after a certain number of cleavages, the embryo attains a particular nuclear-to-cytoplasmic (N/C) ratio, maternal repressors become sufficiently diluted, and the cell cycle slows down. Here we resolve the frog ZGA in time and space by profiling RNA polymerase II (RNAPII) engagement and its transcriptional readout. We detect a gradual increase in both the quantity and the length of RNAPII elongation before the MBT, revealing that >1,000 zygotic genes disregard the N/C timer for their activation, and that the sizes of newly transcribed genes are not necessarily constrained by cell cycle duration. We also find that Wnt, Nodal and BMP signaling together generate most of the spatio-temporal dynamics of regional ZGA, directing the formation of orthogonal body axes and proportionate germ layers.
]]></description>
<dc:creator>Gentsch, G.</dc:creator>
<dc:creator>Owens, N. D. L.</dc:creator>
<dc:creator>Smith, J. C.</dc:creator>
<dc:date>2018-12-06</dc:date>
<dc:identifier>doi:10.1101/488056</dc:identifier>
<dc:title><![CDATA[The Spatio-Temporal Control of Zygotic Genome Activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/494914v1?rss=1">
<title>
<![CDATA[
Nucleosome positioning stability is a significant modulator of germline mutation rate variation across the human genome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/494914v1?rss=1"
</link>
<description><![CDATA[
Understanding the patterns and genesis of germline de novo mutations is important for studying genome evolution and human diseases. Nucleosome organization is suggested to be a contributing factor to mutation rate variation across the genome. However, the small number of published de novo mutations and the low resolution of earlier nucleosome maps limited our understanding of how nucleosome organization affects germline mutation rates in the human genome. Here, we systematically investigated the relationship between nucleosome organization and fine-scale mutation rate variation by analyzing >300,000 de novo mutations from whole-genome trio sequencing and high-resolution nucleosome maps in human. We found that de novo mutation rates are elevated around strong, translationally stable nucleosomes, a previously under-appreciated aspect. We confirmed this observation having controlled for local sequence context and other potential confounding factors. Analysis of the underlying mutational processes suggests that the increased mutation rates around strong nucleosomes are shaped by a combination of low-fidelity replication, frequent DNA damage and insufficient/error-prone repair in these regions. Interestingly, strong nucleosomes are preferentially located in young SINE/LINE elements, implying frequent nucleosome re-positioning (i.e. shifting of dyad position) and their contribution to hypermutation at new retrotransposons during evolution. These findings provide novel insights into how chromatin organization affects germline mutation rates and have important implications in human genetics and genome evolution.
]]></description>
<dc:creator>Li, C.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:date>2018-12-13</dc:date>
<dc:identifier>doi:10.1101/494914</dc:identifier>
<dc:title><![CDATA[Nucleosome positioning stability is a significant modulator of germline mutation rate variation across the human genome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/496851v1?rss=1">
<title>
<![CDATA[
Interleukin-36 promotes systemic Type-I IFN responses in severe psoriasis. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/496851v1?rss=1"
</link>
<description><![CDATA[
Psoriasis is an immune-mediated skin disorder associated with severe systemic co-morbidities. Both chronic and acute forms of the disease are characterised by abnormal interleukin (IL)-36 signalling. While the mechanisms whereby IL-36 promotes cutaneous inflammation are well established, its systemic effects have not been investigated. To address this issue, we initially measured leukocyte gene expression in generalised pustular psoriasis, an acute disease variant caused by mutations of the IL-36 receptor antagonist. By undertaking whole-blood and neutrophil RNA-sequencing in affected individuals, we identified a Type-I IFN signature, which correlated with IL-36 signalling up-regulation. We then validated these observations in patients with chronic plaque psoriasis. Finally, we demonstrated that IL-36 acts directly on plasmacytoid dendritic cells, where it potentiates Toll-like Receptor (TLR)-9 activation and IFN production. This effect was mediated by the induction of PLSCR1, an endosomal TLR-9 transporter. These results define an IL-36/TLR-9/Type-I IFN axis that could be targeted for the treatment of psoriasis co-morbidities.
]]></description>
<dc:creator>Catapano, M.</dc:creator>
<dc:creator>Vergnano, M.</dc:creator>
<dc:creator>Romano, M.</dc:creator>
<dc:creator>Mahil, S. K.</dc:creator>
<dc:creator>Choon, S.-E.</dc:creator>
<dc:creator>Burden, D. A.</dc:creator>
<dc:creator>Young, H. S.</dc:creator>
<dc:creator>Carr, I. M.</dc:creator>
<dc:creator>Lachmann, H. J.</dc:creator>
<dc:creator>Lombardi, G.</dc:creator>
<dc:creator>Smith, C.</dc:creator>
<dc:creator>Ciccarelli, F. D.</dc:creator>
<dc:creator>Barker, J. N.</dc:creator>
<dc:creator>Capon, F.</dc:creator>
<dc:date>2018-12-14</dc:date>
<dc:identifier>doi:10.1101/496851</dc:identifier>
<dc:title><![CDATA[Interleukin-36 promotes systemic Type-I IFN responses in severe psoriasis.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/499244v1?rss=1">
<title>
<![CDATA[
Using deep maxout neural networks to improve the accuracy of function prediction from protein interaction networks 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/499244v1?rss=1"
</link>
<description><![CDATA[
Protein-protein interaction network data provides valuable information that infers direct links between genes and their biological roles. This information brings a fundamental hypothesis for protein function prediction that interacting proteins tend to have similar functions. With the help of recently-developed network embedding feature generation methods and deep maxout neural networks, it is possible to extract functional representations that encode direct links between protein-protein interactions information and protein function. Our novel method, STRING2GO, successfully adopts deep maxout neural networks to learn functional representations simultaneously encoding both protein-protein interactions and functional predictive information. The experimental results show that STRING2GO outperforms other network embedding-based prediction methods and one benchmark method adopted in a recent large scale protein function prediction competition.
]]></description>
<dc:creator>Wan, C.</dc:creator>
<dc:creator>Cozzetto, D.</dc:creator>
<dc:creator>Fa, R.</dc:creator>
<dc:creator>Jones, D. T.</dc:creator>
<dc:date>2018-12-17</dc:date>
<dc:identifier>doi:10.1101/499244</dc:identifier>
<dc:title><![CDATA[Using deep maxout neural networks to improve the accuracy of function prediction from protein interaction networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/498865v1?rss=1">
<title>
<![CDATA[
TorC2 dependent phosphorylation modulates calcium regulation of fission yeast myosin. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/498865v1?rss=1"
</link>
<description><![CDATA[
All cells have the ability to respond to changes in their environment. Signalling networks modulate cytoskeleton and membrane organisation to impact cell cycle progression, polarised cell growth and multicellular development according to the environmental setting. Using diverse in vitro, in vivo and single molecule techniques we have explored the role of myosin-1 signalling in regulating endocytosis during both mitotic and meiotic cell cycles. We have established that a conserved serine within the neck region of the sole fission yeast myosin-1 is phosphorylated in a TORC2 dependent manner to modulate myosin function. Myo1 neck phosphorylation brings about a change in the conformation of the neck region and modifies its interaction with calmodulins, Myo1 dynamics at endocytic foci, and promotes calcium dependent switching between different calmodulin light chains. These data provide insight into a novel mechanism by which myosin neck phosphorylation modulates acto-myosin dynamics to control polarised cell growth in response to mitotic and meiotic cell-cycle progression and the cellular environment.
]]></description>
<dc:creator>Baker, K. B.</dc:creator>
<dc:creator>Gyamfi, I. A.</dc:creator>
<dc:creator>Mashanov, G. I.</dc:creator>
<dc:creator>Molloy, J. E.</dc:creator>
<dc:creator>Geeves, M. A.</dc:creator>
<dc:creator>Mulvihill, D. P.</dc:creator>
<dc:date>2018-12-18</dc:date>
<dc:identifier>doi:10.1101/498865</dc:identifier>
<dc:title><![CDATA[TorC2 dependent phosphorylation modulates calcium regulation of fission yeast myosin.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/499806v1?rss=1">
<title>
<![CDATA[
CSynth: A Dynamic Modelling and Visualisation Tool for 3D Chromatin Structure 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/499806v1?rss=1"
</link>
<description><![CDATA[
The 3D structure of chromatin in the nucleus is important for gene expression and regulation. Chromosomal conformation capture techniques, such as Hi-C, generate large amounts of data showing interaction points on the genome but these are hard to interpret using standard tools. We have developed CSynth, a high performance 3D genome browser and real time chromatin restraint-based modeller to visualise dynamic and interactive models of chromatin capture data. CSynth does its calculations in the GPU hence is much faster than existing modelling software to infer and visualise the chromatin structure which also allow real-time interaction with the modelling parameters. It also allows straightforward comparison of interaction data and the results of third party 3D modelling outputs. In addition we include an option to view and manipulate these complicated structures using Virtual Reality (VR) allowing scientists to immerse themselves in the models for further understanding. This VR component has also proven to be a valuable teaching and public engagement tool. CSynth is web based and available to use at http://csynth.org.
]]></description>
<dc:creator>Todd, S.</dc:creator>
<dc:creator>Todd, P.</dc:creator>
<dc:creator>McGowan, S. J.</dc:creator>
<dc:creator>Hughes, J. R.</dc:creator>
<dc:creator>Kakui, Y.</dc:creator>
<dc:creator>Leymarie, F. F.</dc:creator>
<dc:creator>Latham, W.</dc:creator>
<dc:creator>Taylor, S.</dc:creator>
<dc:date>2019-01-03</dc:date>
<dc:identifier>doi:10.1101/499806</dc:identifier>
<dc:title><![CDATA[CSynth: A Dynamic Modelling and Visualisation Tool for 3D Chromatin Structure]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/513457v1?rss=1">
<title>
<![CDATA[
Whole Genome Doubling mitigates Muller’s Ratchet in Cancer Evolution 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/513457v1?rss=1"
</link>
<description><![CDATA[
Whole genome doubling (WGD) is a prevalent macro-evolutionary event in cancer, involving a doubling of the entire chromosome complement. However, despite its prevalence and clinical prognostic relevance, the evolutionary selection pressures for WGD have not been investigated. Here, we explored whether WGD may act to mitigate the irreversible, inexorable ratchet-like, accumulation of deleterious mutations in essential genes. Utilizing 1050 tumor regions from 816 non-small cell lung cancers (NSCLC), we temporally dissect mutations to determine their temporal acquisition in relation to WGD. We find evidence for strong negative selection against homozygous loss of essential cancer genes prior to WGD. However, mutations in essential genes occurring after duplication were not subject to significant negative selection, consistent with WGD providing a buffering effect, decreasing the likelihood of homozygous loss. Finally, we demonstrate that loss of heterozygosity and temporal dissection of mutations can be exploited to identify signals of positive selection in lung, breast, colorectal cancer and other cancer types, enabling the elucidation of novel tumour suppressor genes and a deeper characterization of known cancer genes.
]]></description>
<dc:creator>Lopez, S.</dc:creator>
<dc:creator>Lim, E.</dc:creator>
<dc:creator>Huebner, A.</dc:creator>
<dc:creator>Dietzen, M.</dc:creator>
<dc:creator>Mourikis, T.</dc:creator>
<dc:creator>Watkins, T. B.</dc:creator>
<dc:creator>Rowan, A.</dc:creator>
<dc:creator>Dewhurst, S. M.</dc:creator>
<dc:creator>Birkbak, N. J.</dc:creator>
<dc:creator>Wilson, G. A.</dc:creator>
<dc:creator>Jamal-Hanjani, M.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>TRACERx Consortium,</dc:creator>
<dc:creator>McGranahan, N.</dc:creator>
<dc:date>2019-01-07</dc:date>
<dc:identifier>doi:10.1101/513457</dc:identifier>
<dc:title><![CDATA[Whole Genome Doubling mitigates Muller’s Ratchet in Cancer Evolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/514166v1?rss=1">
<title>
<![CDATA[
Comparative analysis of Synthetic Physical Interactions with the yeast centrosome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/514166v1?rss=1"
</link>
<description><![CDATA[
1The yeast centrosome or Spindle Pole Body (SPB) is situated in the nuclear membrane, where it nucleates spindle microtubules and acts as a signalling hub. Previously, we used Synthetic Physical Interactions to map the regions of the cell that are sensitive to forced relocalization of proteins across the proteome [Berry et al., 2016]. Here, we expand on this work to show that the SPB, in particular, is sensitive to the relocalization of many proteins. This work inspired a new data analysis approach that indicates that relocalization screens may produce more growth defects than previously reported. A set of associations with the SPB result in elevated SPB number and since hyper-proliferation of centrosomes is a hallmark of cancer cells, these associations point the way for the use of yeast models in the study of spindle formation and chromosome segregation in cancer.
]]></description>
<dc:creator>Howell, R. S.</dc:creator>
<dc:creator>Csikasz-Nagy, A.</dc:creator>
<dc:creator>Thorpe, P. H.</dc:creator>
<dc:date>2019-01-07</dc:date>
<dc:identifier>doi:10.1101/514166</dc:identifier>
<dc:title><![CDATA[Comparative analysis of Synthetic Physical Interactions with the yeast centrosome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/512921v1?rss=1">
<title>
<![CDATA[
Genomic approaches reveal an endemic sub-population of gray wolves in Southern China 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/512921v1?rss=1"
</link>
<description><![CDATA[
Despite being one of the most widely distributed terrestrial mammals, the history of gray wolves (Canis lupus) in China is not well understood as their habitats have been destroyed with growing economic development. Using six specimens from wolf skins in Chinese Natural History museums, we sequenced their genome using a modified ancient DNA procedure. Using whole genome sequence analysis, we showed that gray wolves from Southern China (SC) derive from a single lineage, distinct from gray wolves from the Tibetan Plateau (Canis lupus chanco) and Northern China, suggesting that SC gray wolves may form a distinct sub-population. Of SC gray wolves, one wolf from Zhejiang carries a genetic component from a canid that must have diverged earlier from other wolves than jackals did, perhaps through gene flow from a population related to or further diverged from wolves than the dhole, a species distributed in Southern China and Southeast Asia. This may indicate that interspecific gene flow likely played an important role in shaping the speciation patterns and population structure in the genus Canis. Our study is the first to survey museum genomes of gray wolves from Southern China, revealing the presence of an endemic population with ancient interspecific gene flow from a population related to the dhole, and highlighting how sequencing the paleogenome from museum specimens can help us to study extinct species.
]]></description>
<dc:creator>Wang, G.-D.</dc:creator>
<dc:creator>Zhang, M.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Yang, M. A.</dc:creator>
<dc:creator>Cao, P.</dc:creator>
<dc:creator>Liu, F.</dc:creator>
<dc:creator>Lu, H.</dc:creator>
<dc:creator>Feng, X.</dc:creator>
<dc:creator>Skoglund, P.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Fu, Q.</dc:creator>
<dc:creator>Zhang, Y.-P.</dc:creator>
<dc:date>2019-01-08</dc:date>
<dc:identifier>doi:10.1101/512921</dc:identifier>
<dc:title><![CDATA[Genomic approaches reveal an endemic sub-population of gray wolves in Southern China]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/512764v1?rss=1">
<title>
<![CDATA[
Missense variants in health and disease target distinct functional pathways and proteomics features 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/512764v1?rss=1"
</link>
<description><![CDATA[
1Missense variants are present amongst the healthy population, but some of them are causative of human diseases. Therefore, a classification of variants associated with "healthy" or "diseased" states is not always straightforward. A deeper understanding of the nature of missense variants in health and disease, the cellular processes they may affect, and the general molecular principles which underlie these differences, is essential to better distinguish pathogenic from population variants. Here we quantify variant enrichment across full-length proteins, their domains and 3D-structure defined regions. We integrate this with available transcriptomic and proteomic (protein half-life, thermal stability, abundance) data. Using this approach we have mined a rich set of molecular features which enable us to understand the differences underlying pathogenic and population variants: pathogenic variants mainly affect proteins involved in cell proliferation and nucleotide processing, localise to protein cores and interaction interfaces, and are enriched in more abundant proteins. In terms of their molecular properties, we find that common population variants and pathogenic variants show the greatest contrast. Additionally, in contrary to other studies, we find that rare population variants display features closer to common than pathogenic variants. This study provides molecular details into how different proteins exhibit resilience and/or sensitivity towards missense variants. Such details could be harnessed to predict variant deleteriousness, and prioritise variant-enriched proteins and protein domains for therapeutic targeting and development. The ZoomVar database, which we created for this study, is available at http://fraternalilab.kcl.ac.uk/ZoomVar. It allows users to programmatically annotate a large number of missense variants with protein structural information, and to calculate variant enrichment in different protein structural regions.nnSignificance StatementOne of the greatest challenges in understanding the genetic basis of diseases is to discriminate between likely harmless and potentially disease-causing sequence variants. To better evaluate the pathogenic potential of missense variants, we developed a strategy to quantitatively measure the enrichment of both disease and non disease-related variants within a protein based on its structural and domain organisation. By integrating available transcriptomics and proteomics data, our approach distinguishes pathogenic from population variants far more clearly than previously possible, and reveals hitherto unknown details of how different proteins exhibit resilience and/or sensitivity towards genetic variants. Our results will help to prioritise variant-enriched proteins for therapeutic targeting; we have created the ZoomVar database, accessible at http://fraternalilab.kcl.ac.uk/ZoomVar, for programmatic mapping of user-defined variants to protein structural and domain information.
]]></description>
<dc:creator>Laddach, A.</dc:creator>
<dc:creator>Ng, J. C.-F.</dc:creator>
<dc:creator>Fraternali, F.</dc:creator>
<dc:date>2019-01-09</dc:date>
<dc:identifier>doi:10.1101/512764</dc:identifier>
<dc:title><![CDATA[Missense variants in health and disease target distinct functional pathways and proteomics features]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/525709v1?rss=1">
<title>
<![CDATA[
A Novel Repertoire of Blood Transcriptome Modules Based on Co-expression Patterns Across Sixteen Disease and Physiological States 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/525709v1?rss=1"
</link>
<description><![CDATA[
As the capacity for generating large scale data continues to grow the ability to extract meaningful biological knowledge from it remains a limitation. Here we describe the development of a new fixed repertoire of transcriptional modules. It is meant to serve as a stable reusable framework for the analysis and interpretation of blood transcriptome profiling data. It is supported by customized resources, which include analysis workflows, fingerprint grid plots data visualizations, interactive web applications providing access to a vast number of module-specific functional profiling reports, reference transcriptional profiles and give users the ability to visualize of changes in transcript abundance across the modular repertoire at different granularity levels. A use case focusing on a set of six modules comprising interferon-inducible genes is also provided. Altogether we hope that this resource will also serve as a framework for improving over time our collective understanding of the immunobiology underlying blood transcriptome profiling data.
]]></description>
<dc:creator>Altman, M. C.</dc:creator>
<dc:creator>Rinchai, D.</dc:creator>
<dc:creator>Baldwin, N.</dc:creator>
<dc:creator>Whalen, E.</dc:creator>
<dc:creator>Garand, M.</dc:creator>
<dc:creator>Kabeer, B. A.</dc:creator>
<dc:creator>Toufiq, M.</dc:creator>
<dc:creator>Presnell, S.</dc:creator>
<dc:creator>Chiche, L.</dc:creator>
<dc:creator>Jourde-Chiche, N.</dc:creator>
<dc:creator>Phillips, T.</dc:creator>
<dc:creator>Klintmalm, G.</dc:creator>
<dc:creator>O'Garra, A.</dc:creator>
<dc:creator>Berry, M.</dc:creator>
<dc:creator>Bloom, C.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Graham, C. M.</dc:creator>
<dc:creator>Lipman, M.</dc:creator>
<dc:creator>Lertmemongkolchai, G.</dc:creator>
<dc:creator>Bedognetti, D.</dc:creator>
<dc:creator>Kheradmand, F.</dc:creator>
<dc:creator>Mejias, A.</dc:creator>
<dc:creator>Ramilo, O.</dc:creator>
<dc:creator>Palucka, K.</dc:creator>
<dc:creator>Pascual, V.</dc:creator>
<dc:creator>Banchereau, J.</dc:creator>
<dc:creator>Chaussabel, D.</dc:creator>
<dc:date>2019-01-23</dc:date>
<dc:identifier>doi:10.1101/525709</dc:identifier>
<dc:title><![CDATA[A Novel Repertoire of Blood Transcriptome Modules Based on Co-expression Patterns Across Sixteen Disease and Physiological States]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/527564v1?rss=1">
<title>
<![CDATA[
The visual system of the genetically tractable crustacean Parhyale hawaiensis: diversification of eyes and visual circuits associated with low-resolution vision 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/527564v1?rss=1"
</link>
<description><![CDATA[
BackgroundArthropod eyes have diversified during evolution to serve multiple needs, such as finding mates, hunting prey, and navigating in complex surroundings under varying light conditions. This diversity is reflected in the optical apparatus, photoreceptors and neural circuits that underpin vision. While this diversity has been extensively documented, our ability to genetically manipulate the visual system to investigate its function is largely limited to a single species, the fruitfly Drosophila melanogaster. Here, we describe the visual system of Parhyale hawaiensis, an amphipod crustacean for which we have established tailored genetic tools.

ResultsAdult Parhyale have apposition-type compound eyes made up of [~]50 ommatidia. Each ommatidium contains four photoreceptor cells with large rhabdomeres (R1-4), expected to be sensitive to the polarisation of light, and one photoreceptor cell with a smaller rhabdomere (R5). The two types of photoreceptors express different opsins, belonging to families with distinct wavelength sensitivities. Using the cis.-regulatory regions of opsin genes, we established transgenic reporters expressed in each photoreceptor cell type. Based on these reporters, we show that R1-4 and R5 photoreceptors extend axons to the first optic lobe neuropil, revealing striking differences compared with the photoreceptor projections found in related crustaceans and insects. Investigating visual function, we show that Parhyale has a positive phototactic response and is capable of adapting its eyes to different levels of light intensity.

ConclusionsWe propose that the visual system of Parhyale serves low-resolution visual tasks, such as orientation and navigation, based on broad gradients of light intensity and polarisation. Optic lobe structure and photoreceptor projections point to significant divergence from the conserved visual circuits found in other malacostracan crustaceans and insects, which could be associated with a shift to low-resolution vision. Our study provides the foundation for research in the visual system of this genetically tractable species.
]]></description>
<dc:creator>Ramos, A. P.</dc:creator>
<dc:creator>Gustafsson, O.</dc:creator>
<dc:creator>Labert, N.</dc:creator>
<dc:creator>Salecker, I.</dc:creator>
<dc:creator>Nilsson, D. E.</dc:creator>
<dc:creator>Averof, M.</dc:creator>
<dc:date>2019-01-23</dc:date>
<dc:identifier>doi:10.1101/527564</dc:identifier>
<dc:title><![CDATA[The visual system of the genetically tractable crustacean Parhyale hawaiensis: diversification of eyes and visual circuits associated with low-resolution vision]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/536342v1?rss=1">
<title>
<![CDATA[
Neuronal differentiation affects tissue mechanics and progenitor arrangement in the vertebrate neuroepithelium 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/536342v1?rss=1"
</link>
<description><![CDATA[
Cell division, movement and differentiation contribute to pattern formation in developing tissues. This is the case in the vertebrate neural tube where neurons differentiate in a characteristic pattern from a highly dynamic proliferating pseudostratified epithelium. To investigate how progenitor proliferation and differentiation affect cell arrangement and growth of the neural tube, we use experimental measurements to develop a mechanical model of the apical surface of the neuroepithelium that incorporates inter-kinetic nuclear movement and spatially varying rates of neuronal differentiation. Simulations predict that tissue growth and the shape of lineage-related clones of cells differ with the rate of differentiation. Growth is isotropic in regions of high differentiation, but dorsoventrally biased in regions of low differentiation. This is consistent with experimental observations. The absence of directional signalling in the simulations indicates that global mechanical constraints are sufficient to explain the observed differences in anisotropy. This provides insight into how the tissue growth rate affects cell dynamics and growth anisotropy and opens up possibilities to study the coupling between mechanics, pattern formation and growth in the neural tube.
]]></description>
<dc:creator>Guerrero, P.</dc:creator>
<dc:creator>Perez-Carrasco, R.</dc:creator>
<dc:creator>Zagorski, M.</dc:creator>
<dc:creator>Page, D.</dc:creator>
<dc:creator>Kicheva, A.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Page, K.</dc:creator>
<dc:date>2019-01-31</dc:date>
<dc:identifier>doi:10.1101/536342</dc:identifier>
<dc:title><![CDATA[Neuronal differentiation affects tissue mechanics and progenitor arrangement in the vertebrate neuroepithelium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/541615v1?rss=1">
<title>
<![CDATA[
Simultaneous tracking of cell motility in liquid and at the solid-liquid interface 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/541615v1?rss=1"
</link>
<description><![CDATA[
1.To better understand key behaviors of living cells, such as bacterial biofilm formation, they must be observed above surfaces and at the interface between the surface and liquid medium. We have established a methodology for label-free imaging and tracking of individual cells simultaneously at both the solid-liquid interface and within the bulk, utilizing imaging modes of digital holographic microscopy (DHM) in 3D, differential interference contrast (DIC) and total internal reflectance microscopy (TIRM) in 2D as well as analysis protocols using a bespoke software package. We illustrate the power of this method by making detailed single cell measurements of Pseudomonas aeruginosa in the first minutes of their interaction with a glass surface, focusing on the role of the flagella stators, motAB and motCD. Using this new method we have determined their relative contributions to bulk and near surface motion for populations of cells at the single cell level.
]]></description>
<dc:creator>Hook, A. L.</dc:creator>
<dc:creator>Flewellen, J. L.</dc:creator>
<dc:creator>Zaid, I. M.</dc:creator>
<dc:creator>Berry, R. M.</dc:creator>
<dc:creator>Dubern, J.-F.</dc:creator>
<dc:creator>Carabelli, A.</dc:creator>
<dc:creator>Wildman, R.</dc:creator>
<dc:creator>Russell, N.</dc:creator>
<dc:creator>Williams, P.</dc:creator>
<dc:creator>Alexander, M. R.</dc:creator>
<dc:date>2019-02-05</dc:date>
<dc:identifier>doi:10.1101/541615</dc:identifier>
<dc:title><![CDATA[Simultaneous tracking of cell motility in liquid and at the solid-liquid interface]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/541235v1?rss=1">
<title>
<![CDATA[
Segment 2 from influenza A(H1N1)pdm09 viruses confers temperature sensitive HA yield on candidate vaccine virus growth in eggs that is complemented by PB2 701D 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/541235v1?rss=1"
</link>
<description><![CDATA[
Candidate vaccine viruses (CVVs) for seasonal influenza A virus are made by reassortment of the antigenic virus with a high-yielding egg-adapted strain, typically A/Puerto Rico/8/34 (PR8). Many 2009 H1N1 pandemic (pdm09) high-growth reassortants (HGRs) selected by this process contain pdm09 segment 2 in addition to the antigenic genes. To investigate this, we made CVV mimics by reverse genetics (RG) that were either 6:2 or 5:3 reassortants between PR8 and two pdm09 strains, A/California/7/2009 (Cal7) and A/England/195/2009, differing in the source of segment 2. The 5:3 viruses replicated better in MDCK-SIAT1 cells than the 6:2 viruses, but the 6:2 CVVs gave higher HA antigen yields from eggs. This unexpected phenomenon reflected temperature sensitivity conferred by pdm09 segment 2, as HA yields from eggs for the 5:3 viruses improved substantially when viruses were grown at 35{degrees}C compared with 37.5{degrees}C, whereas 6:2 virus yield did not. Authentic 5:3 pdm09 HGRs, X-179A and X-181, were not markedly temperature-sensitive however, despite their PB1 sequences being identical to that of Cal7, suggestive of compensatory mutations elsewhere in the genome. Sequence comparisons of the PR8-derived backbone genes identified single changes in PB2 and NP, 5 in NS1, and 1 in NS2. PB2 N701D but not NP T130A affected the temperature dependency of viral transcription. Furthermore, introducing the PB2 701D change into a 5:3 CVV mimic improved and drastically reduced the temperature sensitivity of HA yield. We conclude that RG PR8 backbones used for vaccine manufacture in eggs should contain PB2 701D to maximise virus yield.
]]></description>
<dc:creator>Hussain, S.</dc:creator>
<dc:creator>Turnbull, M. L.</dc:creator>
<dc:creator>Pinto, R. M.</dc:creator>
<dc:creator>McCauley, J. W.</dc:creator>
<dc:creator>Engelhardt, O. G.</dc:creator>
<dc:creator>Digard, P.</dc:creator>
<dc:date>2019-02-08</dc:date>
<dc:identifier>doi:10.1101/541235</dc:identifier>
<dc:title><![CDATA[Segment 2 from influenza A(H1N1)pdm09 viruses confers temperature sensitive HA yield on candidate vaccine virus growth in eggs that is complemented by PB2 701D]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/553602v1?rss=1">
<title>
<![CDATA[
Perturbation of the gut microbiota by antibiotics results in accelerated breast tumour growth and metabolic dysregulation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/553602v1?rss=1"
</link>
<description><![CDATA[
BackgroundBreast cancer is the second most prevalent cancer worldwide with around 1.7 million new cases diagnosed every year. Whilst prognosis is generally favourable in early stages, this worsens significantly in advanced disease. Therefore, it is pertinent to focus on mitigating factors that may slow growth or progression. Recently, the gut microbiome has been implicated in a wide-range of roles in tumour biology. Through modulation of immunity, the gut microbiota can improve the efficacy of several immunotherapies. However, despite the prevalence of breast cancer, there is still a lack of microbiota studies in this field, including exploring the influence of external microbiome-modulating factors such as antibiotics. We describe herein how disruption of the gut microbiota via antibiotics may be detrimental to patient outcomes through acceleration of tumour growth.

ResultsSupplementing animals with a cocktail of antibiotics leads to gut microbiota alterations and is accompanied by significant acceleration of tumour growth. Surprisingly, and distinct from previous microbiome-tumour studies, the mechanism driving these effects do not appear to be due to gross immunological changes. Analysis of intratumoural immune cell populations and cytokine production are not affected by antibiotic administration. Through global tumour transcriptomics, we have uncovered dysregulated gene expression networks relating to protein and lipid metabolism that are correlated with accelerated tumour growth. Fecal metabolomics revealed a reduction of the microbial-derived short-chain fatty acid butyrate that may contribute to accelerated tumour growth. Finally, through use of a routinely administered antibiotic in breast cancer patients, Cephalexin, we have shown that tumour growth is also significantly affected. Metataxanomic sequencing and analysis highlighted significant antibiotic-associated reductions in the butyrate producing genera Odoribacter and Anaeotruncus, and increased abundance of Bacteroides.

ConclusionsOur data indicate that perturbation of the microbiota by antibiotics may have negative impacts on breast cancer patient outcomes. This is of importance as antibiotics are regularly prescribed to breast cancer patients undergoing mastectomy or breast reconstruction. We have also shown that the metabolic impact of disruption to the microbiome should be considered alongside the potent immunological effects. We believe our work lays the foundation for improving the use of antibiotics in patients, and with further investigation could potentially inform clinical practice.
]]></description>
<dc:creator>Kirkup, B.</dc:creator>
<dc:creator>McKee, A.</dc:creator>
<dc:creator>Makin, K.</dc:creator>
<dc:creator>Paveley, J.</dc:creator>
<dc:creator>Caim, S.</dc:creator>
<dc:creator>Alcon-Giner, C.</dc:creator>
<dc:creator>Leclaire, C.</dc:creator>
<dc:creator>Dalby, M.</dc:creator>
<dc:creator>Le Gall, G.</dc:creator>
<dc:creator>Andrusaite, A.</dc:creator>
<dc:creator>Kreuzaler, P.</dc:creator>
<dc:creator>Driscoll, P.</dc:creator>
<dc:creator>MacRae, J.</dc:creator>
<dc:creator>Calvani, E.</dc:creator>
<dc:creator>Ghanate, A.</dc:creator>
<dc:creator>Milling, S.</dc:creator>
<dc:creator>Yuneva, M.</dc:creator>
<dc:creator>Weilbaecher, K.</dc:creator>
<dc:creator>Korcsmaros, T.</dc:creator>
<dc:creator>Hall, L. J.</dc:creator>
<dc:creator>Robinson, S. D.</dc:creator>
<dc:date>2019-02-19</dc:date>
<dc:identifier>doi:10.1101/553602</dc:identifier>
<dc:title><![CDATA[Perturbation of the gut microbiota by antibiotics results in accelerated breast tumour growth and metabolic dysregulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/555995v1?rss=1">
<title>
<![CDATA[
Improving diagnosis and monitoring of treatment response in pulmonary tuberculosis using the molecular bacterial load assay (MBLA) 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/555995v1?rss=1"
</link>
<description><![CDATA[
ObjectivesBetter outcomes in tuberculosis require new diagnostic and treatment monitoring tools. In this paper we evaluated the utility of a marker of M. tuberculosis viable count, the Molecular Bacterial Load assay (MBLA) for diagnosis and treatment monitoring of tuberculosis in a high burden setting.

MethodsPatients with smear positive pulmonary tuberculosis from two sites in Tanzania and one each in Malawi and Mozambique. Sputum samples were taken weekly for the first 12 weeks of treatment and evaluated by MBLA and mycobacterial growth indicator tube method (MGIT).

ResultsThe results of high and low positive control samples confirmed inter site reproducibility. Over the 12 weeks of treatment there was a steady decline in the viable bacterial load as measured by the MBLA that corresponds to rise in time to a positive result (TTP) in the Mycobacterial Growth Indicator Tube. Both MBLA and MGIT provided similar time to test negativity. Importantly, as treatment progressed samples in MGIT were increasingly likely to be contaminated, which compromised the acquisition of results but did not affect MBLA samples.

ConclusionsMBLA produces a reproducible measure of Mtb viable count comparable to that of MGIT that is not compromised by contamination in a real-world setting. As a molecular test, the results can be available in as little as four hours and could allow health care professionals to identify rapidly patients who are failing therapy.
]]></description>
<dc:creator>Sabiiti, W.</dc:creator>
<dc:creator>Azam, K.</dc:creator>
<dc:creator>Kuchaka, D.</dc:creator>
<dc:creator>Mtafya, B.</dc:creator>
<dc:creator>Bowness, R. E.</dc:creator>
<dc:creator>Oravcova, K.</dc:creator>
<dc:creator>Farmer, E.</dc:creator>
<dc:creator>Honeyborne, I.</dc:creator>
<dc:creator>Evangelopoulos, D.</dc:creator>
<dc:creator>McHugh, T. D.</dc:creator>
<dc:creator>Xiao, H.</dc:creator>
<dc:creator>Khosa, C.</dc:creator>
<dc:creator>Rachow, A.</dc:creator>
<dc:creator>Heinrich, N.</dc:creator>
<dc:creator>Kampira, E.</dc:creator>
<dc:creator>Davies, G.</dc:creator>
<dc:creator>Bhatt, N.</dc:creator>
<dc:creator>Ntinginya, N.</dc:creator>
<dc:creator>Viegas, S.</dc:creator>
<dc:creator>Jani, I.</dc:creator>
<dc:creator>Kamdolozi, M.</dc:creator>
<dc:creator>Mdolo, a.</dc:creator>
<dc:creator>Khonga, M.</dc:creator>
<dc:creator>Boeree, M. J.</dc:creator>
<dc:creator>Philips, P. P.</dc:creator>
<dc:creator>Sloan, D. J.</dc:creator>
<dc:creator>Hoelscher, M.</dc:creator>
<dc:creator>Kibiki, G.</dc:creator>
<dc:creator>Gillespie, S. H.</dc:creator>
<dc:date>2019-02-28</dc:date>
<dc:identifier>doi:10.1101/555995</dc:identifier>
<dc:title><![CDATA[Improving diagnosis and monitoring of treatment response in pulmonary tuberculosis using the molecular bacterial load assay (MBLA)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/565416v1?rss=1">
<title>
<![CDATA[
TGF-β family ligands exhibit distinct signaling dynamics that are driven by receptor localization 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/565416v1?rss=1"
</link>
<description><![CDATA[
Growth factor-induced signal transduction pathways are tightly regulated at multiple points intracellularly, but how cells monitor levels of extracellular ligand and translate this information into appropriate downstream responses remains unclear. Understanding signaling dynamics is thus a key challenge in determining how cells respond to external cues. Here, we demonstrate that different TGF-{beta} family ligands, namely Activin A and BMP4, signal with distinct dynamics, which differ profoundly from those of TGF-{beta} itself. The distinct signaling dynamics are driven by differences in the localization and internalization of receptors for each ligand, which in turn determine the capability of cells to monitor levels of extracellular ligand. Using mathematical modeling, we demonstrate that the distinct receptor behaviors and signaling dynamics observed may be primarily driven by differences in ligand-receptor affinity. Furthermore, our results provide a clear rationale for the different mechanisms of pathway regulation found in vivo for each of these growth factors.
]]></description>
<dc:creator>Miller, D. S. J.</dc:creator>
<dc:creator>Schmierer, B.</dc:creator>
<dc:creator>Hill, C. S.</dc:creator>
<dc:date>2019-03-01</dc:date>
<dc:identifier>doi:10.1101/565416</dc:identifier>
<dc:title><![CDATA[TGF-β family ligands exhibit distinct signaling dynamics that are driven by receptor localization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/566430v1?rss=1">
<title>
<![CDATA[
A Complex Interplay of Anionic Phospholipid Binding Regulates 3-Phosphoinositide-Dependent-Kinase-1 Homodimer Activation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/566430v1?rss=1"
</link>
<description><![CDATA[
3-Phosphoinositide-dependent-Kinase-1 is a master regulator whereby its PI3-kinase-dependent dysregulation in human pathologies is well documented. Understanding the direct role for PtdIns(3,4,5)P3 and other anionic phospholipids in the regulation of PDK1 conformational dynamics and its downstream activation remains incomplete.

Using advanced quantitative-time-resolved imaging, FCS and molecular modelling, we show an interplay of antagonistic binding effects of PtdIns(3,4,5)P3 and other anionic phospholipids, regulating activated PDK1 homodimers. We demonstrate that phosphatidylserine maintains PDK1 in an inactive conformation. The dysregulation of the PI3K pathway affects the spatio-temporal and conformational dynamics of PDK1 and the activation of its downstream substrates.

We establish an anionic-phospholipid-dependent model for PDK1 regulation, depicting the conformational dynamics of multiple homodimer states. The dysregulation of the PI3K pathway perturbs equilibrium between the PDK1 homodimer conformations. Our findings indicate that the alteration of specific basic residues of PDK1-PH domain leads to its constitutive activation, a potential significance in different types of carcinomas.
]]></description>
<dc:creator>De Las Heras, G.</dc:creator>
<dc:creator>Calleja, V.</dc:creator>
<dc:creator>Bailly, R.</dc:creator>
<dc:creator>Dessolin, J.</dc:creator>
<dc:creator>Requejo-Isidro, J.</dc:creator>
<dc:creator>Larijani, B.</dc:creator>
<dc:date>2019-03-03</dc:date>
<dc:identifier>doi:10.1101/566430</dc:identifier>
<dc:title><![CDATA[A Complex Interplay of Anionic Phospholipid Binding Regulates 3-Phosphoinositide-Dependent-Kinase-1 Homodimer Activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/566711v1?rss=1">
<title>
<![CDATA[
The Personal Genome Project-UK: an open access resource of human multi-omics data 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/566711v1?rss=1"
</link>
<description><![CDATA[
Integrative analysis of multi-omics data is a powerful approach for gaining functional insights into biological and medical processes. Conducting these multifaceted analyses on human samples is often complicated by the fact that the raw sequencing output is rarely available under open access. The Personal Genome Project UK (PGP-UK) is one of few resources that recruits its participants under open consent and makes the resulting multi-omics data freely and openly available. As part of this resource, we describe the PGP-UK multi-omics reference panel consisting of ten genomic, methylomic and transcriptomic data. Specifically, we outline the data processing, quality control and validation procedures which were implemented to ensure data integrity and exclude sample mix-ups. In addition, we provide a REST API to facilitate the download of the entire PGP-UK dataset. The data are also available from two cloud-based environments, providing platforms for free integrated analysis. In conclusion, the genotype-validated PGP-UK multi-omics human reference panel described here provides a valuable new open access resource for integrated analyses in support of personal and medical genomics.
]]></description>
<dc:creator>Chervova, O.</dc:creator>
<dc:creator>Conde, L.</dc:creator>
<dc:creator>Guerra-Assuncao, J. A.</dc:creator>
<dc:creator>Moghul, I.</dc:creator>
<dc:creator>Webster, A. P.</dc:creator>
<dc:creator>Berner, A.</dc:creator>
<dc:creator>Larose Cadieux, E.</dc:creator>
<dc:creator>Tian, Y.</dc:creator>
<dc:creator>Voloshin, V.</dc:creator>
<dc:creator>Hamoudi, R. A.</dc:creator>
<dc:creator>Herrero, J.</dc:creator>
<dc:creator>Beck, S.</dc:creator>
<dc:date>2019-03-04</dc:date>
<dc:identifier>doi:10.1101/566711</dc:identifier>
<dc:title><![CDATA[The Personal Genome Project-UK: an open access resource of human multi-omics data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/563304v1?rss=1">
<title>
<![CDATA[
Identification of RBPMS as a smooth muscle master splicing regulator via proximity of its gene with super-enhancers 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/563304v1?rss=1"
</link>
<description><![CDATA[
Alternative splicing (AS) programs are primarily controlled by regulatory RNA binding proteins (RBPs). It has been proposed that a small number of master splicing regulators might control cell-specific splicing networks and that these RBPs could be identified by proximity of their genes to transcriptional super-enhancers. Using this approach we identified RBPMS as a critical splicing regulator in differentiated vascular smooth muscle cells (SMCs). RBPMS is highly down-regulated during phenotypic switching of SMCs from a contractile to a motile and proliferative phenotype and is responsible for 20% of the AS changes during this transition. RBPMS directly regulates AS of numerous components of the actin cytoskeleton and focal adhesion machineries whose activity is critical for SMC function in both phenotypes. RBPMS also regulates splicing of other splicing, post-transcriptional and transcription regulators including the key SMC transcription factor Myocardin, thereby matching many of the criteria of a master regulator of AS in SMCs.
]]></description>
<dc:creator>Nakagaki-Silva, E. E.</dc:creator>
<dc:creator>Gooding, C.</dc:creator>
<dc:creator>Llorian, M.</dc:creator>
<dc:creator>Jacob, A. G.</dc:creator>
<dc:creator>Richards, F.</dc:creator>
<dc:creator>Buckroyd, A.</dc:creator>
<dc:creator>Sinha, S.</dc:creator>
<dc:creator>Smith, C. W.</dc:creator>
<dc:date>2019-02-28</dc:date>
<dc:identifier>doi:10.1101/563304</dc:identifier>
<dc:title><![CDATA[Identification of RBPMS as a smooth muscle master splicing regulator via proximity of its gene with super-enhancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/568527v1?rss=1">
<title>
<![CDATA[
An Integrated Chemical Proteomics Approach for Quantitative Profiling of Intracellular ADP-Ribosylation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/568527v1?rss=1"
</link>
<description><![CDATA[
ADP-ribosylation is integral to a diverse range of cellular processes such as DNA repair, chromatin regulation and RNA processing. However, proteome-wide investigation of its cellular functions has been limited due to numerous technical challenges including the complexity of the poly(ADP-ribose) (PAR) chains, low abundance of the modification and lack of sensitive enrichment methods. To facilitate live-cell profiling of ADP-ribosylated proteins, we show that 2-alkyne-adenosine (2YnAd) is metabolically incorporated in mammalian cells and enables fluorescent detection and robust affinity enrichment of the modified proteins. We then present an integrated chemical biology approach that involves simultaneous metabolic incorporation of 2YnAd and the previously reported 6-alkyne-adenosine (6YnAd) in live cells followed by click chemistry with a capture reagent to facilitate highly sensitive and comprehensive enrichment of the modified proteins. By combining this dual metabolic labelling strategy with the tandem mass tag (TMT) isobaric mass spectrometry, we have quantified the responses of thousands of endogenous proteins to clinical PARP inhibitors Olaparib and Rucaparib. Our study provides insight into the wider scope of PARP targets and will help to advance further characterisation of their functional roles.
]]></description>
<dc:creator>Kalesh, K.</dc:creator>
<dc:creator>Lukauskas, S.</dc:creator>
<dc:creator>Borg, A. J.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Leung, A. K. L.</dc:creator>
<dc:creator>Haskard, D. O.</dc:creator>
<dc:creator>DiMaggio, P. A.</dc:creator>
<dc:date>2019-03-05</dc:date>
<dc:identifier>doi:10.1101/568527</dc:identifier>
<dc:title><![CDATA[An Integrated Chemical Proteomics Approach for Quantitative Profiling of Intracellular ADP-Ribosylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/568691v1?rss=1">
<title>
<![CDATA[
In the chick embryo, estrogen can induce chromosomally male ZZ left gonad epithelial cells to form an ovarian cortex which supports oogenesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/568691v1?rss=1"
</link>
<description><![CDATA[
In chickens, as in most amniotes, the first morphogenetic event in ovary differentiation is the formation of two distinct domains: a steroidogenic core, the medulla, overlain by the germ cell niche, the cortex. This process normally starts immediately after sex determination in the ZW embryos, substantially before the progression of germ cells into meiosis. In order to address the extent to which the cortical domain depends on intrinsic or extrinsic factors, we generated models of gonadal intersex by mixing ZW (female) and ZZ (male) cells in gonadal chimeras, or by altering estrogen levels of ZZ and ZW embryos in ovo. We found that both chomosomally female and male cells can be induced to form the cortical domain and that this can form relatively independently of the phenotypic sex of the medulla as long as estrogen is provided. We also show that the cortex promoting activity of estrogen signalling is mediated via Estrogen Receptor alpha within the left gonad epithelium. Therefore, either a ZW or ZZ cortical domain can provide an adequate niche to allow the germ cells to progress into meiosis. However, the presence of a medulla with an "intersex" or male phenotype may compromise this progression, causing cortical germ cells to remain in an immature state in the embryo.
]]></description>
<dc:creator>Guioli, S.</dc:creator>
<dc:creator>Zhao, D.</dc:creator>
<dc:creator>Nandi, S.</dc:creator>
<dc:creator>Clinton, M.</dc:creator>
<dc:creator>Lovell-Badge, R.</dc:creator>
<dc:date>2019-03-05</dc:date>
<dc:identifier>doi:10.1101/568691</dc:identifier>
<dc:title><![CDATA[In the chick embryo, estrogen can induce chromosomally male ZZ left gonad epithelial cells to form an ovarian cortex which supports oogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/569368v1?rss=1">
<title>
<![CDATA[
Differential requirements of cyclase associated protein (CAP) for actin turnover during the lytic cycle of Toxoplasma gondii 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/569368v1?rss=1"
</link>
<description><![CDATA[
Toxoplasma gondii contains a limited subset of actin binding proteins. Here we show that ablation of the putative actin regulator cyclase-associated protein (TgCAP) leads to significant defects in some but not all actin dependent processes, including a defect in cell-cell communication, but surprisingly not synchronicity of division. Two CAP isoforms originate from alternative translational start sites and are beneficial for parasite fitness while a single isoform is sufficient for virulence in mice. Examination of the mutant parasites by 3D electron microscopy reveals that loss of CAP results in a defect to form a normal residual body, but all parasites remain connected within the vacuole. This dissociates synchronicity of division and parasite rosetting and reveals that establishment and maintenance of the residual body may be more complex than previously thought. These results highlight the different spatial requirements for actin turnover in Toxoplasma, controlled by a reduced subset of actin binding proteins.
]]></description>
<dc:creator>Hunt, A.</dc:creator>
<dc:creator>Wagener, J.</dc:creator>
<dc:creator>Kent, R.</dc:creator>
<dc:creator>Carmeille, R.</dc:creator>
<dc:creator>Russell, M.</dc:creator>
<dc:creator>Peddie, C.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:creator>Heaslip, A.</dc:creator>
<dc:creator>Ward, G. E.</dc:creator>
<dc:creator>Treeck, M.</dc:creator>
<dc:date>2019-03-06</dc:date>
<dc:identifier>doi:10.1101/569368</dc:identifier>
<dc:title><![CDATA[Differential requirements of cyclase associated protein (CAP) for actin turnover during the lytic cycle of Toxoplasma gondii]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/570069v1?rss=1">
<title>
<![CDATA[
CHIME: CMOS-hosted in-vivo microelectrodes for massively scalable neuronal recordings 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/570069v1?rss=1"
</link>
<description><![CDATA[
Mammalian brains consist of 10s of millions to 100s of billions of neurons operating at millisecond time scales, of which current recording techniques only capture a tiny fraction. Recording techniques capable of sampling neural activity at such temporal resolution have been difficult to scale: The most intensively studied mammalian neuronal networks, such as the neocortex, show layered architecture, where the optimal recording technology samples densely over large areas. However, the need for application-specific designs as well as the mismatch between the threedimensional architecture of the brain and largely two-dimensional microfabrication techniques profoundly limits both neurophysiological research and neural prosthetics.nnHere, we propose a novel strategy for scalable neuronal recording by combining bundles of glass-ensheathed microwires with large-scale amplifier arrays derived from commercial CMOS of in-vitro MEA systems or high-speed infrared cameras. High signal-to-noise ratio (<20 V RMS noise floor, SNR up to 25) is achieved due to the high conductivity of core metals in glass-ensheathed microwires allowing for ultrathin metal cores (down to <1 m) and negligible stray capacitance. Multi-step electrochemical modification of the tip enables ultra-low access impedance with minimal geometric area and largely independent of core diameter. We show that microwire size can be reduced to virtually eliminate damage to the blood-brain-barrier upon insertion and demonstrate that microwire arrays can stably record single unit activity.nnCombining microwire bundles and CMOS arrays allows for a highly scalable neuronal recording approach, linking the progress of electrical neuronal recording to the rapid scaling of silicon microfabrication. The modular design of the system allows for custom arrangement of recording sites. Our approach of employing bundles of minimally invasive, highly insulated and functionalized microwires to lift a 2-dimensional CMOS architecture into the 3rd dimension can be translated to other CMOS arrays such as electrical stimulation devices.
]]></description>
<dc:creator>Kollo, M.</dc:creator>
<dc:creator>Racz, R. R.</dc:creator>
<dc:creator>Hanna, M.</dc:creator>
<dc:creator>Obaid, A.</dc:creator>
<dc:creator>Angle, M. R.</dc:creator>
<dc:creator>Wray, W.</dc:creator>
<dc:creator>Kong, Y.</dc:creator>
<dc:creator>Hierlemann, A.</dc:creator>
<dc:creator>Muller, J.</dc:creator>
<dc:creator>Melosh, N. A.</dc:creator>
<dc:creator>Schaefer, A. T.</dc:creator>
<dc:date>2019-03-08</dc:date>
<dc:identifier>doi:10.1101/570069</dc:identifier>
<dc:title><![CDATA[CHIME: CMOS-hosted in-vivo microelectrodes for massively scalable neuronal recordings]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/571885v1?rss=1">
<title>
<![CDATA[
Oncogenic signaling alters cell shape and mechanics to facilitate cell division under confinement. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/571885v1?rss=1"
</link>
<description><![CDATA[
When cells enter mitosis, they become spherical and mechanically stiffen. We used MCF10A cell lines as a model system in which to investigate the effect of induced oncogene expression on mitotic entry. We find that activation of oncogenic RasV12, for as little as five hours, changes the way cells divide. RasV12-dependent activation of the MEK-ERK signalling cascade alters acto-myosin contractility to enhance mitotic rounding. RasV12 also affects cell mechanics, so that RasV12 expressing cells are softer in interphase but stiffen more upon entry into mitosis. As a consequence, RasV12 expression augments the ability of cells to round up and divide faithfully when confined underneath a stiff hydrogel. Conversely, inhibition of the Ras-ERK pathway reduces mitotic rounding under confinement, resulting in chromosome segregation defects. These data suggest a novel mechanism by which oncogenic Ras-ERK signalling can aid division in stiff environments like those found in tumours.
]]></description>
<dc:creator>Matthews, H. K.</dc:creator>
<dc:creator>Ganguli, S.</dc:creator>
<dc:creator>Plak, K.</dc:creator>
<dc:creator>Taubenberger, A. V.</dc:creator>
<dc:creator>Piel, M.</dc:creator>
<dc:creator>Guck, J.</dc:creator>
<dc:creator>Baum, B.</dc:creator>
<dc:date>2019-03-09</dc:date>
<dc:identifier>doi:10.1101/571885</dc:identifier>
<dc:title><![CDATA[Oncogenic signaling alters cell shape and mechanics to facilitate cell division under confinement.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/570689v1?rss=1">
<title>
<![CDATA[
Mammalian olfaction is a high temporal bandwidth sense 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/570689v1?rss=1"
</link>
<description><![CDATA[
Odours are transported in turbulent plumes resulting locally in highly fluctuating odour concentration (Celani et al., 2014; Murlis et al., 1992; Mylne and Mason, 1991; Shraiman and Siggia, 2000). Yet, whether mammals can make use of the ensuing temporal structure (Celani et al., 2014; Crimaldi and Koseff, 2001; Murlis et al., 1992; Mylne and Mason, 1991; Schmuker et al., 2016; Vickers, 2000) to extract information about the olfactory environment remains unknown. Here, we use dual-energy photoionisation recording with >300 Hz bandwidth to simultaneously determine odour concentrations of two odours in air. We show that temporal correlation of odour concentrations reliably predicts whether odorants emerge from the same or different sources in normal turbulent environments outside and in laboratory conditions. To replicate natural odour dynamics in a reproducible manner we developed a multichannel odour delivery device allowing presentation of several odours with 10 ms temporal resolution. Integrating this device in an automated operant conditioning system we demonstrate that mice can reliably discriminate the correlation structure of odours at frequencies of up to 40 Hz. Consistent with this finding, output neurons in the olfactory bulb show segregated responses depending on the correlation of odour stimuli with populations of 10s of neurons sufficient to reach behavioural performance. Our work thus demonstrates that mammals can perceive temporal structure in odour stimuli at surprisingly fast timescales. This in turn might be useful for key behavioural challenges (Jacobs, 2012) such as odour source separation (Hopfield, 1991), figure-ground separation (Rokni et al., 2014) or odour localisation (Vergassola et al., 2007; Vickers, 2000).
]]></description>
<dc:creator>Erskine, A.</dc:creator>
<dc:creator>Ackels, T.</dc:creator>
<dc:creator>Dasgupta, D.</dc:creator>
<dc:creator>Fukunaga, I.</dc:creator>
<dc:creator>Schaefer, A. T.</dc:creator>
<dc:date>2019-03-09</dc:date>
<dc:identifier>doi:10.1101/570689</dc:identifier>
<dc:title><![CDATA[Mammalian olfaction is a high temporal bandwidth sense]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/571745v1?rss=1">
<title>
<![CDATA[
Lactate potentiates differentiation and expansion of cytotoxic T cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/571745v1?rss=1"
</link>
<description><![CDATA[
Exercise has a range of effects on metabolism. In animal models, repeated exertion reduces malignant tumour progression, and clinically, exercise can improve outcome for cancer patients. The etiology of the effect of exercise on tumour progression is unclear, as are the cellular actors involved. We show here that exercise-induced reduction in tumour growth is dependent on CD8+ T cells and that lactate, which is produced at high levels during exertion, increases proliferative capacity and cytotoxicity of CD8+ T cells. We found that at elevated levels lactate is used as a fuel during T cell activation. We further found that injection of lactate into animals can reduce malignant tumour growth in a dose-and CD8+ T cell-dependent manner. These data demonstrate that lactate can act to increase the anti-tumour activity of cytotoxic T cells, and in so doing, reduce cancer progression.
]]></description>
<dc:creator>Rundqvist, H.</dc:creator>
<dc:creator>Velica, P.</dc:creator>
<dc:creator>Barbieri, L.</dc:creator>
<dc:creator>Gameiro, P.</dc:creator>
<dc:creator>Cunha, P.</dc:creator>
<dc:creator>Gojkovic, M.</dc:creator>
<dc:creator>Mijwel, S.</dc:creator>
<dc:creator>Ahlstedt, E.</dc:creator>
<dc:creator>Foskolou, I. P.</dc:creator>
<dc:creator>Ruiz-Perez, M.</dc:creator>
<dc:creator>Arsenian-Henriksson, M.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Ostman, A.</dc:creator>
<dc:creator>Johnson, R.</dc:creator>
<dc:date>2019-03-09</dc:date>
<dc:identifier>doi:10.1101/571745</dc:identifier>
<dc:title><![CDATA[Lactate potentiates differentiation and expansion of cytotoxic T cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/573295v1?rss=1">
<title>
<![CDATA[
Massively Parallel Microwire Arrays Integrated with CMOS chips for Neural Recording 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/573295v1?rss=1"
</link>
<description><![CDATA[
Multi-channel electrical recordings of neural activity in the brain is an increasingly powerful method revealing new aspects of neural communication, computation, and prosthetics. However, while planar silicon-based CMOS devices in conventional electronics scale rapidly, neural interface devices have not kept pace. Here, we present a new strategy to interface silicon-based chips with three-dimensional microwire arrays, providing the link between rapidly-developing electronics and high density neural interfaces. The system consists of a bundle of microwires mated to large-scale microelectrode arrays, such as camera chips. This system has excellent recording performance, demonstrated via single unit and local-field potential recordings in isolated retina and in the motor cortex or striatum of awake moving mice. The modular design enables a variety of microwire types and sizes to be integrated with different types of pixel arrays, connecting the rapid progress of commercial multiplexing, digitisation and data acquisition hardware together with a three-dimensional neural interface.
]]></description>
<dc:creator>Obaid, A. M.</dc:creator>
<dc:creator>Hanna, M.-E. S.</dc:creator>
<dc:creator>Wu, Y.-W.</dc:creator>
<dc:creator>Kollo, M.</dc:creator>
<dc:creator>Racz, R. R.</dc:creator>
<dc:creator>Angle, M. R.</dc:creator>
<dc:creator>Muller, J.</dc:creator>
<dc:creator>Brackbill, N.</dc:creator>
<dc:creator>Wray, W.</dc:creator>
<dc:creator>Franke, F.</dc:creator>
<dc:creator>Chichilnisky, E. J.</dc:creator>
<dc:creator>Hierlemann, A.</dc:creator>
<dc:creator>Ding, J. B.</dc:creator>
<dc:creator>Schaefer, A. T.</dc:creator>
<dc:creator>Melosh, N. A.</dc:creator>
<dc:date>2019-03-11</dc:date>
<dc:identifier>doi:10.1101/573295</dc:identifier>
<dc:title><![CDATA[Massively Parallel Microwire Arrays Integrated with CMOS chips for Neural Recording]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/576660v1?rss=1">
<title>
<![CDATA[
Pax9 is required for cardiovascular development and interacts with Tbx1 in the pharyngeal endoderm to control 4th pharyngeal arch artery morphogenesis. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/576660v1?rss=1"
</link>
<description><![CDATA[
Developmental defects affecting the heart and aortic arch arteries are a key phenotype observed in DiGeorge syndrome patients and are caused by a microdeletion on chromosome 22q11. Heterozygosity of TBX1, one of the deleted genes, is expressed throughout the pharyngeal arches and is considered a key component for the arch artery defects. Pax9 is expressed in the pharyngeal endoderm and is downregulated in Tbx1 mutant mice. We show here that Pax9 deficient mice are born with complex cardiovascular malformations affecting the outflow tract and aortic arch arteries with failure of the 3rd and 4th pharyngeal arch arteries to form correctly. Transcriptome analysis indicated that Pax9 and Tbx1 may function together, and mice double heterozygous for Tbx1/Pax9 presented with a significantly increased incidence of interrupted aortic arch when compared to Tbx1 heterozygous mice. Using a novel Pax9Cre allele we demonstrated that the site of this Tbx1-Pax9 genetic interaction is in the pharyngeal endoderm, therefore revealing that a Tbx1/Pax9-controlled signalling mechanism emanating from the pharyngeal endoderm is required for critical tissue interactions during normal morphogenesis of the pharyngeal arch artery system.nnSummary statementPax9 is required for outflow tract and aortic arch development, and functions together with Tbx1 in the pharyngeal endoderm for 4th arch artery formation.
]]></description>
<dc:creator>Phillips, H. M.</dc:creator>
<dc:creator>Stothard, C. A.</dc:creator>
<dc:creator>Shaikh Qureshi, W. M.</dc:creator>
<dc:creator>Kousa, A. I.</dc:creator>
<dc:creator>Briones-Leon, A.</dc:creator>
<dc:creator>Khasawneh, R.</dc:creator>
<dc:creator>Sanders, R.</dc:creator>
<dc:creator>Mazotta, S.</dc:creator>
<dc:creator>Dodds, R.</dc:creator>
<dc:creator>Seidel, K.</dc:creator>
<dc:creator>Bates, T.</dc:creator>
<dc:creator>Nakatomi, M.</dc:creator>
<dc:creator>Cockell, S.</dc:creator>
<dc:creator>Schneider, J. E.</dc:creator>
<dc:creator>Mohun, T. J.</dc:creator>
<dc:creator>Maehr, R.</dc:creator>
<dc:creator>Kist, R.</dc:creator>
<dc:creator>Peters, H.</dc:creator>
<dc:creator>Bamforth, S. D.</dc:creator>
<dc:date>2019-03-14</dc:date>
<dc:identifier>doi:10.1101/576660</dc:identifier>
<dc:title><![CDATA[Pax9 is required for cardiovascular development and interacts with Tbx1 in the pharyngeal endoderm to control 4th pharyngeal arch artery morphogenesis.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/579250v1?rss=1">
<title>
<![CDATA[
Neuronal programming by microbiota enables environmental regulation of intestinal motility 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/579250v1?rss=1"
</link>
<description><![CDATA[
Environmental signals modulate the activity of the nervous system and harmonize its output with the outside world. Synaptic activity is crucial for integrating sensory and effector neural pathways but the role of transcriptional mechanisms as environmental sensors in the nervous system remains unclear. By combining a novel strategy for transcriptomic profiling of enteric neurons with microbiota manipulation, we demonstrate that the transcriptional programs of intestinal neural circuits depend on their anatomical and physiological context. We also identify the ligand-dependent transcription factor Aryl hydrocarbon Receptor (AhR) is an intrinsic regulator of enteric nervous system output. AhR is instated as a neuronal biosensor in response to microbiota colonization allowing resident enteric neurons to directly monitor and respond to the intestinal microenvironment. We suggest that AhR signaling integrates neuronal activity with host defence mechanisms towards gut homeostasis and health.nnOne Sentence SummaryMicrobiota induce expression of AhR in enteric neurons of the distal intestine enabling them to respond to environmental signals.
]]></description>
<dc:creator>Obata, Y.</dc:creator>
<dc:creator>Boeing, S.</dc:creator>
<dc:creator>Castano, A.</dc:creator>
<dc:creator>Carina Bon-Frauches, A.</dc:creator>
<dc:creator>Gomez de Aguero, M.</dc:creator>
<dc:creator>Boesmans, W.</dc:creator>
<dc:creator>Yilmaz, B.</dc:creator>
<dc:creator>Lopes, R.</dc:creator>
<dc:creator>Huseynova, A.</dc:creator>
<dc:creator>Maradana, M. R.</dc:creator>
<dc:creator>Vanden Berghe, P.</dc:creator>
<dc:creator>Murray, A. J.</dc:creator>
<dc:creator>Stockinger, B.</dc:creator>
<dc:creator>Macpherson, A. J.</dc:creator>
<dc:creator>Pachnis, V.</dc:creator>
<dc:date>2019-03-15</dc:date>
<dc:identifier>doi:10.1101/579250</dc:identifier>
<dc:title><![CDATA[Neuronal programming by microbiota enables environmental regulation of intestinal motility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/579805v1?rss=1">
<title>
<![CDATA[
ESCRT-III/Vps4 controls heterochromatin-nuclear envelope attachments 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/579805v1?rss=1"
</link>
<description><![CDATA[
In eukaryotes chromosomes are compartmentalized within the nucleus delimited by the double membrane of the nuclear envelope (NE). Defects in the function and structure of the NE are linked to disease1,2. During interphase, the NE organizes the genome and regulates its expression3. As cells enter mitosis, chromosomes are released from the NE, which is then remodelled to form the daughter nuclei at mitotic exit4. Interactions between the NE and chromatin underpinning both interphase and post-mitotic NE functions are executed by inner nuclear membrane (INM) proteins such as members of the evolutionarily conserved chromatin-binding LEM-domain family5-8. How chromatin tethering by these transmembrane proteins is controlled in interphase and if such a regulation contributes to subsequent NE dynamics in mitosis remains unclear. Here we probe these fundamental questions using an emerging model organism, the fission yeast Schizosaccharomyces japonicus, which breaks and reforms the NE during mitosis9,10. We show that attachments between heterochromatin and the transmembrane Lem2-Nur1 complex are continuously remodelled in interphase by the ESCRT-III/AAA-ATPase Vps4 machinery. ESCRT-III/Vps4 mediates the release of Lem2-Nur1 from heterochromatin as a prerequisite for the timely progression through mitosis. Failure in this process leads to persistent association of chromosomes with the INM, which prevents Lem2-Nur1 from re-localizing to the sites of NE sealing around the mitotic spindle and severely delays re-establishment of nucleocytoplasmic compartmentalization. Our work establishes the INM transmembrane Lem2-Nur1 complex as a  substrate for ESCRT-III/Vps4 to couple dynamic tethering of chromosomes to the INM with the establishment of nuclear compartmentalization.
]]></description>
<dc:creator>Pieper, G. H.</dc:creator>
<dc:creator>Sprenger, S.</dc:creator>
<dc:creator>Teis, D.</dc:creator>
<dc:creator>Oliferenko, S.</dc:creator>
<dc:date>2019-03-15</dc:date>
<dc:identifier>doi:10.1101/579805</dc:identifier>
<dc:title><![CDATA[ESCRT-III/Vps4 controls heterochromatin-nuclear envelope attachments]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/582056v1?rss=1">
<title>
<![CDATA[
Protein structure without structure determination: direct coupling analysis based on in vitro evolution 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/582056v1?rss=1"
</link>
<description><![CDATA[
Protein structure is tightly inter-twined with function according to the laws of evolution. Understanding how structure determines function has been the aim of structural biology for decades. Here, we have wondered instead whether it is possible to exploit the function for which a protein was evolutionary selected to gain information on protein structure and on the landscape explored during the early stages of molecular and natural evolution. To answer to this question, we developed a new methodology, which we named CAMELS (Coupling Analysis by Molecular Evolution Library Sequencing), that is able to obtain the in vitro evolution of a protein from an artificial selection based on function. We were able to observe with CAMELS many features of the TEM-1 beta lactamase local fold exclusively by generating and sequencing large libraries of mutational variants. We demonstrated that we can, whenever a functional phenotypic selection of a protein is available, sketch the structural and evolutionary landscape of a protein without utilizing purified proteins, collecting physical measurements or relying on the pool of natural protein variants.
]]></description>
<dc:creator>Fantini, M.</dc:creator>
<dc:creator>Lisi, S.</dc:creator>
<dc:creator>de los Rios, P.</dc:creator>
<dc:creator>Cattaneo, A.</dc:creator>
<dc:creator>Pastore, A.</dc:creator>
<dc:date>2019-03-20</dc:date>
<dc:identifier>doi:10.1101/582056</dc:identifier>
<dc:title><![CDATA[Protein structure without structure determination: direct coupling analysis based on in vitro evolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/583674v1?rss=1">
<title>
<![CDATA[
Identification of reduced host transcriptomic signatures for tuberculosis and digital PCR-based validation and quantification 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/583674v1?rss=1"
</link>
<description><![CDATA[
Recently, host whole blood gene expression signatures have been identified for diagnosis of tuberculosis (TB). Absolute quantification of the concentrations of signature transcripts in blood have not been reported, but would facilitate the development of diagnostic tests.nnTo identify minimal transcript signatures, we applied a novel transcript selection procedure to microarray data from African adults comprising 536 patients with TB, other diseases (OD) and latent TB (LTBI), divided into training and test sets. Signatures were validated using reverse transcriptase (RT) - digital PCR (dPCR).nnA four-transcript signature (GBP6, TMCC1, PRDM1, ARG1) measured using RT-dPCR distinguished TB patients from those with OD (area under the curve (AUC) 93.8% (CI95% 82.2 - 100%). A three-transcript signature (FCGR1A, ZNF296, C1QB) differentiated TB from LTBI (AUC 97.3%, CI95%: 93.3 - 100%), regardless of HIV.nnThese signatures have been validated across platforms and across samples offering strong, quantitative support for their use as diagnostic biomarkers for TB.
]]></description>
<dc:creator>Gliddon, H. D.</dc:creator>
<dc:creator>Kaforou, M.</dc:creator>
<dc:creator>Alikian, M.</dc:creator>
<dc:creator>Habgood-Coote, D.</dc:creator>
<dc:creator>Zhou, C.</dc:creator>
<dc:creator>Oni, T.</dc:creator>
<dc:creator>Anderson, S. T.</dc:creator>
<dc:creator>Brent, A. J.</dc:creator>
<dc:creator>Crampin, A. C.</dc:creator>
<dc:creator>Eley, B.</dc:creator>
<dc:creator>Heyderman, R. S.</dc:creator>
<dc:creator>Kern, F.</dc:creator>
<dc:creator>Langford, P. R.</dc:creator>
<dc:creator>Ottenhoff, T. H.</dc:creator>
<dc:creator>French, N.</dc:creator>
<dc:creator>Hibberd, M. L.</dc:creator>
<dc:creator>Wright, V. J.</dc:creator>
<dc:creator>Dockrell, H. M.</dc:creator>
<dc:creator>Coin, L. J.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Levin, M.</dc:creator>
<dc:date>2019-03-21</dc:date>
<dc:identifier>doi:10.1101/583674</dc:identifier>
<dc:title><![CDATA[Identification of reduced host transcriptomic signatures for tuberculosis and digital PCR-based validation and quantification]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/586719v1?rss=1">
<title>
<![CDATA[
ERK1/2 signalling dynamics promote neural differentiation by regulating the polycomb repressive complex 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/586719v1?rss=1"
</link>
<description><![CDATA[
Fibroblast Growth Factor (FGF) is a neural inducer in many vertebrate embryos, but how it regulates chromatin organization to coordinate the activation of neural genes is unclear. Moreover, for differentiation to progress FGF signalling has to decline. Why this signalling dynamic is required has not been determined. Here we show that dephosphorylation of the FGF effector kinase ERK1/2 rapidly increases chromatin accessibility at neural genes in mouse embryos and, using ATAC-seq in human embryonic stem cell derived spinal cord precursors, we demonstrate that this occurs across hundreds of neural genes. Importantly, while Erk1/2 inhibition induces precocious neural gene transcription, this step involves dissociation of the polycomb repressive complex from gene loci and takes places independently of subsequent loss of the repressive histone mark H3K27me3 and transcriptional onset. We find that loss of ERK1/2 activity but not its occupancy at neural genes is critical for this mechanism. Moreover, transient ERK1/2 inhibition is sufficient for polycomb protein dissociation and this is not reversed on resumption of ERK1/2 signalling. These data indicate that ERK1/2 signalling maintains polycomb repressive complexes at neural genes, that its decline coordinates their increased accessibility and that this is a directional molecular mechanism, which initiates the process of neural commitment. Furthermore, as the polycomb repressive complexes repress but also ready genes for transcription, these findings suggest that ERK1/2 promotion of these complexes is a rite of passage for subsequent differentiation.
]]></description>
<dc:creator>Semprich, C. I.</dc:creator>
<dc:creator>Metzis, V.</dc:creator>
<dc:creator>Patel, H.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Storey, K. G.</dc:creator>
<dc:date>2019-03-22</dc:date>
<dc:identifier>doi:10.1101/586719</dc:identifier>
<dc:title><![CDATA[ERK1/2 signalling dynamics promote neural differentiation by regulating the polycomb repressive complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/586800v1?rss=1">
<title>
<![CDATA[
Prediction of inter-residue contacts with DeepMetaPSICOV in CASP13 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/586800v1?rss=1"
</link>
<description><![CDATA[
In this article, we describe our efforts in contact prediction in the CASP13 experiment. We employed a new deep learning-based contact prediction tool, DeepMetaPSICOV (or DMP for short), together with new methods and data sources for alignment generation. DMP evolved from MetaPSICOV and DeepCov and combines the input feature sets used by these methods as input to a deep, fully convolutional residual neural network. We also improved our method for multiple sequence alignment generation and included metagenomic sequences in the search. We discuss successes and failures of our approach and identify areas where further improvements may be possible. DMP is freely available at: https://github.com/psipred/DeepMetaPSICOV.
]]></description>
<dc:creator>Kandathil, S. M.</dc:creator>
<dc:creator>Greener, J. G.</dc:creator>
<dc:creator>Jones, D. T.</dc:creator>
<dc:date>2019-03-24</dc:date>
<dc:identifier>doi:10.1101/586800</dc:identifier>
<dc:title><![CDATA[Prediction of inter-residue contacts with DeepMetaPSICOV in CASP13]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/587295v1?rss=1">
<title>
<![CDATA[
A Transcriptome Fingerprinting Assay for Clinical Immune Monitoring 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/587295v1?rss=1"
</link>
<description><![CDATA[
BackgroundWhile our understanding of the role that the immune system plays in health and disease is growing at a rapid pace, available clinical tools to capture this complexity are lagging. We previously described the construction of a third-generation modular transcriptional repertoire derived from genome-wide transcriptional profiling of blood of 985 subjects across 16 diverse immunologic conditions, which comprises 382 distinct modules.nnResultsHere we describe the use of this modular repertoire framework for the development of a targeted transcriptome fingerprinting assay (TFA). The first step consisted in down-selection of the number of modules to 32, on the basis of similarities in changes in transcript abundance and functional interpretation. Next down-selection took place at the level of each of the 32 modules, with each one of them being represented by four transcripts in the final 128 gene panel. The assay was implemented on both the Fluidigm high throughput microfluidics PCR platform and the Nanostring platform, with the list of assays target probes being provided for both. Finally, we provide evidence of the versatility of this assay to assess numerous immune functions in vivo by demonstrating applications in the context of disease activity assessment in systemic lupus erythematosus and longitudinal immune monitoring during pregnancy.nnConclusionsThis work demonstrates the utility of data-driven network analysis applied to large-scale transcriptional profiling to identify key markers of immune responses, which can be downscaled to a rapid, inexpensive, and highly versatile assay of global immune function applicable to diverse investigations of immunopathogenesis and biomarker discovery.
]]></description>
<dc:creator>Altman, M. C.</dc:creator>
<dc:creator>Baldwin, N.</dc:creator>
<dc:creator>Whalen, E.</dc:creator>
<dc:creator>Al-Shaikhly, T.</dc:creator>
<dc:creator>Presnell, S.</dc:creator>
<dc:creator>Khaenam, P.</dc:creator>
<dc:creator>Gersuk, V. H.</dc:creator>
<dc:creator>Chiche, L.</dc:creator>
<dc:creator>Jourde-Chiche, N.</dc:creator>
<dc:creator>Phillips, T.</dc:creator>
<dc:creator>Klintmalm, G.</dc:creator>
<dc:creator>O'Garra, A.</dc:creator>
<dc:creator>Berry, M.</dc:creator>
<dc:creator>Bloom, C.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Graham, C. M.</dc:creator>
<dc:creator>Lipman, M.</dc:creator>
<dc:creator>Lertmemongkolchai, G.</dc:creator>
<dc:creator>Kheradmand, F.</dc:creator>
<dc:creator>Mejias, A.</dc:creator>
<dc:creator>Ramilo, O.</dc:creator>
<dc:creator>Palucka, K.</dc:creator>
<dc:creator>Pascual, V.</dc:creator>
<dc:creator>Banchereau, J.</dc:creator>
<dc:creator>Chaussabel, D.</dc:creator>
<dc:date>2019-03-24</dc:date>
<dc:identifier>doi:10.1101/587295</dc:identifier>
<dc:title><![CDATA[A Transcriptome Fingerprinting Assay for Clinical Immune Monitoring]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/592311v1?rss=1">
<title>
<![CDATA[
Apical transport of Crumbs maintains epithelial cell polarity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/592311v1?rss=1"
</link>
<description><![CDATA[
Crumbs (Crb in Drosophila; CRB1-3 in mammals) is a transmembrane determinant of epithelial cell polarity and a regulator of Hippo signalling. Crb is normally localized to apical cell-cell contacts, just above adherens junctions, but how apical trafficking of Crb is regulated in epithelial cells remains unclear. We use the Drosophila follicular epithelium to demonstrate that polarized trafficking of Crb is mediated by transport along microtubules by the motor protein Dynein and along actin filaments by the motor protein Myosin-V (MyoV). Blocking transport of Crb-containing vesicles by Dynein or MyoV leads to accumulation of Crb within Rab11 endosomes, rather than apical delivery. The final steps of Crb delivery and stabilisation at the plasma membrane requires the exocyst complex and three apical FERM domain proteins - Merlin, Moesin and Expanded - whose simultaneous loss disrupts apical localization of Crb. Accordingly, a knock-in deletion of the Crb FERM-binding motif (FBM) also impairs apical localization. Finally, overexpression of Crb challenges this system, creating a sensitized background to identify components involved in cytoskeletal polarization, apical membrane trafficking and stabilisation of Crb at the apical domain.
]]></description>
<dc:creator>Aguilar-Aragon, M.</dc:creator>
<dc:creator>Fletcher, G.</dc:creator>
<dc:creator>Thompson, B. J.</dc:creator>
<dc:date>2019-03-28</dc:date>
<dc:identifier>doi:10.1101/592311</dc:identifier>
<dc:title><![CDATA[Apical transport of Crumbs maintains epithelial cell polarity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/595173v1?rss=1">
<title>
<![CDATA[
Mycobacterium tuberculosis cording in the cytosol of live lymphatic endothelial cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/595173v1?rss=1"
</link>
<description><![CDATA[
The ability of Mycobacterium tuberculosis to form serpentine cords is intrinsically related to its virulence, but specifically how M. tuberculosis cording contributes to pathogenesis remains obscure. We show that several M. tuberculosis clinical isolates form intracellular cords in primary human lymphatic endothelial cells (hLEC) in vitro and also in the lymph nodes of patients with tuberculosis. We identified via RNA-seq a transcriptional programme in hLEC that activates cellular pro-survival and cytosolic surveillance of intracellular pathogens pathways. Consistent with this, cytosolic access of hLEC is required for intracellular M. tuberculosis cording; and cord formation is dependent on the M. tuberculosis ESX-1 type VII secretion system and the mycobacterial lipid PDIM. Finally, we show that M. tuberculosis cording is a novel size-dependent mechanism used by the pathogen to evade xenophagy in the cytosol of endothelial cells. These results provide a mechanism that explains the long-standing association between M. tuberculosis cording and virulence.
]]></description>
<dc:creator>Lerner, T. R.</dc:creator>
<dc:creator>Queval, C. J.</dc:creator>
<dc:creator>Lai, R. P.</dc:creator>
<dc:creator>Russell, M. R. G.</dc:creator>
<dc:creator>Fearns, A.</dc:creator>
<dc:creator>Greenwood, D. J.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Gutierrez, M. G.</dc:creator>
<dc:date>2019-04-01</dc:date>
<dc:identifier>doi:10.1101/595173</dc:identifier>
<dc:title><![CDATA[Mycobacterium tuberculosis cording in the cytosol of live lymphatic endothelial cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/595439v1?rss=1">
<title>
<![CDATA[
Mask family proteins ANKHD1 and ANKRD17 regulate YAP nuclear import, stability and phase separation. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/595439v1?rss=1"
</link>
<description><![CDATA[
The Mask family of multiple ankyrin repeat and KH domain proteins were discovered in Drosophila to promote the activity of the transcriptional coactivator Yorkie (Yki), the sole fly homolog of mammalian YAP (YAP1) and TAZ (WWTR1). The molecular function of Mask, or its mammalian homologs Mask1 (ANKHD1) and Mask2 (ANKRD17), remains unclear. Mask family proteins contain two Ankyrin repeat domains that bind Yki/YAP as well as a conserved nuclear localisation sequence (NLS) and nuclear export sequence (NES), suggesting a role in nucleo-cytoplasmic transport. Here we show that Mask acts to promote nuclear import of Yki, and that addition of an ectopic NLS to Yki is sufficient to bypass the requirement for Mask in Yki-driven tissue growth. Mammalian Mask1/2 proteins also promote nuclear import of YAP, as well as stabilising YAP and driving colloidal phase separation into large liquid droplets. Mask1/2 and YAP normally colocalise in a granular fashion in both nucleus and cytoplasm, and are co-regulated during mechanotransduction. Our results suggest that Mask family proteins promote YAP nuclear import and phase separation to regulate YAP stability and transcriptional activity.
]]></description>
<dc:creator>Sidor, C.</dc:creator>
<dc:creator>Borreguero-Munoz, N.</dc:creator>
<dc:creator>Fletcher, G.</dc:creator>
<dc:creator>Elbediwy, A.</dc:creator>
<dc:creator>Vanyai, H.</dc:creator>
<dc:creator>Guillermin, O.</dc:creator>
<dc:creator>Thompson, B. J.</dc:creator>
<dc:date>2019-04-01</dc:date>
<dc:identifier>doi:10.1101/595439</dc:identifier>
<dc:title><![CDATA[Mask family proteins ANKHD1 and ANKRD17 regulate YAP nuclear import, stability and phase separation.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/599381v1?rss=1">
<title>
<![CDATA[
Elevated LRRK2 and α-synuclein levels in CSF of infectious meningitis patients 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/599381v1?rss=1"
</link>
<description><![CDATA[
Neurodegenerative diseases such as Parkinsons (PD) have a complex aetiology consisting of an interplay of genetic and environmental factors. Inflammation and infection are proposed external factors that trigger disease progression. Tuberculous and cryptococcal meningitis frequently lead to long-term neurological sequelae but their association with the development of PD are unexplored. In this study, we protein profiled the CSF from 76 patients with or without infectious meningitis and found that proteins commonly associated with PD (LRRK2, tau and alpha-synuclein) were significantly elevated, establishing a link between neuroinflammation and infection. Importantly, these findings suggest that LRRK2, tau and alpha-synuclein could represent biomarkers of neuroinflammation.
]]></description>
<dc:creator>Herbst, S.</dc:creator>
<dc:creator>Marais, S.</dc:creator>
<dc:creator>Gutierrez, M. G.</dc:creator>
<dc:creator>Waddell, S. J.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Lai, R. P.</dc:creator>
<dc:date>2019-04-04</dc:date>
<dc:identifier>doi:10.1101/599381</dc:identifier>
<dc:title><![CDATA[Elevated LRRK2 and α-synuclein levels in CSF of infectious meningitis patients]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/600833v1?rss=1">
<title>
<![CDATA[
Canonical NOTCH signaling controls the early progenitor state and emergence of the medullary epithelial lineage in fetal thymus development. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/600833v1?rss=1"
</link>
<description><![CDATA[
Thymus function depends on the epithelial compartment of the thymic stroma. Cortical thymic epithelial cells (cTECs) regulate T cell lineage commitment and positive selection, while medullary (m) TECs impose central tolerance on the T cell repertoire. During thymus organogenesis, these functionally distinct sub-lineages are thought to arise from a common thymic epithelial progenitor cell (TEPC). The mechanisms controlling cTEC and mTEC production from the common TEPC are not however understood. Here, we show that emergence of the earliest mTEC lineage-restricted progenitors requires active NOTCH signaling in progenitor TEC and that, once specified, further mTEC development is NOTCH-independent. In addition, we demonstrate that persistent NOTCH activity favors maintenance of undifferentiated TEPC at the expense of cTEC differentiation. Finally, we uncover a direct interaction between NOTCH and FOXN1, the master regulator of TEC differentiation. These data establish NOTCH as a potent regulator of TEPC and mTEC fate during fetal thymus development and are thus of high relevance to strategies aimed at generating/regenerating functional thymic tissue in vitro and in vivo.
]]></description>
<dc:creator>Liu, D.</dc:creator>
<dc:creator>Kousa, A.</dc:creator>
<dc:creator>O'Neill, K.</dc:creator>
<dc:creator>Guillemot, F.</dc:creator>
<dc:creator>Popis, M.</dc:creator>
<dc:creator>Farley, A. M.</dc:creator>
<dc:creator>Tomlinson, S. R.</dc:creator>
<dc:creator>Ulyanchenko, S.</dc:creator>
<dc:creator>Seymour, P. A.</dc:creator>
<dc:creator>Serup, P.</dc:creator>
<dc:creator>Koch, U.</dc:creator>
<dc:creator>Radtke, F.</dc:creator>
<dc:creator>Blackburn, C. C.</dc:creator>
<dc:date>2019-04-05</dc:date>
<dc:identifier>doi:10.1101/600833</dc:identifier>
<dc:title><![CDATA[Canonical NOTCH signaling controls the early progenitor state and emergence of the medullary epithelial lineage in fetal thymus development.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/491787v1?rss=1">
<title>
<![CDATA[
Widespread FUS mislocalization is a molecular hallmark of ALS 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/491787v1?rss=1"
</link>
<description><![CDATA[
Amyotrophic lateral sclerosis (ALS)-causing mutations clearly implicate ubiquitously expressed and predominantly nuclear RNA binding proteins (RBPs), which form pathological cytoplasmic inclusions in this context. However, the possibility that wild-type RBPs mislocalize without necessarily becoming constituents of ALS cytoplasmic inclusions themselves remains unexplored. We hypothesized that nuclear-to-cytoplasmic mislocalization of the RBP Fused in Sarcoma (FUS), in an unaggregated state, may occur more widely in ALS that previously recognized. To address this hypothesis, we analysed motor neurons (MNs) from an human ALS induced-pluripotent stem cells (iPSC) model caused by the VCP mutation. Additionally, we examined mouse transgenic models and post-mortem tissue from human sporadic ALS cases. We report nuclear-to-cytoplasmic mislocalization of FUS in both VCP-mutation related ALS and, crucially, in sporadic ALS spinal cord tissue from multiple cases. Furthermore, we provide evidence that FUS protein binds to an aberrantly retained intron within the SFPQ transcript, which is exported from the nucleus into the cytoplasm. Collectively, these data support a model for ALS pathogenesis whereby aberrant intron-retention in SFPQ transcripts contributes to FUS mislocalization through their direct interaction and nuclear export. In summary, we report widespread mislocalization of the FUS protein in ALS and propose a putative underlying mechanism for this process.
]]></description>
<dc:creator>Tyzack, G. E.</dc:creator>
<dc:creator>Luisier, R.</dc:creator>
<dc:creator>Taha, D. M.</dc:creator>
<dc:creator>Neeves, J.</dc:creator>
<dc:creator>Modic, M.</dc:creator>
<dc:creator>Mitchell, J. S.</dc:creator>
<dc:creator>Meyer, I.</dc:creator>
<dc:creator>Greensmith, L.</dc:creator>
<dc:creator>Newcombe, J.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:creator>Patani, R.</dc:creator>
<dc:date>2019-04-09</dc:date>
<dc:identifier>doi:10.1101/491787</dc:identifier>
<dc:title><![CDATA[Widespread FUS mislocalization is a molecular hallmark of ALS]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/605808v1?rss=1">
<title>
<![CDATA[
A fence function for adherens junctions in epithelial cell polarity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/605808v1?rss=1"
</link>
<description><![CDATA[
Adherens junctions are a defining feature of all epithelial cells, providing cell-cell adhesion and being essential for cell and tissue morphology. In Drosophila, adherens junctions are concentrated between the apical and basolateral plasma membrane domains, but whether they contribute to apical-basal polarisation itself has been unclear. Here we show that, in the absence of adherens junctions, apical-basal polarity determinants can still segregate into complementary domains, but control of apical versus basolateral domain size is lost. Manipulation of the level of apical or basal polarity determinants in experiments and in computer simulations suggests that junctions provide a moveable diffusion barrier, or fence, that restricts the diffusion of polarity determinants to enable precise domain size control. Movement of adherens junctions in response to mechanical forces during morphogenetic change thus enables spontaneous adjustment of apical versus basolateral domain size as an emergent property of the polarising system.
]]></description>
<dc:creator>Aguilar-Aragon, M.</dc:creator>
<dc:creator>Tournier, A.</dc:creator>
<dc:creator>Thompson, B. J.</dc:creator>
<dc:date>2019-04-11</dc:date>
<dc:identifier>doi:10.1101/605808</dc:identifier>
<dc:title><![CDATA[A fence function for adherens junctions in epithelial cell polarity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/603936v1?rss=1">
<title>
<![CDATA[
Natural hypothalamic circuit dynamics underlying object memorization 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/603936v1?rss=1"
</link>
<description><![CDATA[
Memorizing encountered objects is fundamental for normal life, but the underlying natural brain activity remains poorly understood. The hypothalamus is historically implicated in memory disorders, but whether and how its endogenous real-time activity affects object memorization remains unknown. We found that upon self-initiated object encounters, hypothalamic melanin-concentrating hormone (MCH) neurons emit dynamic, object-encounter-associated signals encoding object novelty. Optosilencing of these signals, performed in closed-loop with object encounters selectively during object memory acquisition, prevented the ability to recognize the previously encountered objects. Optogenetic and chemogenetic connectivity analyses demonstrated that local GAD65 neurons form an inhibitory GAD65[-&gt;]MCH microcircuit that controls the object-encounter-associated MCH cell signals. GAD65 cell optosilencing during object memory acquisition enhanced future object recognition through MCH-receptor-dependent pathways. These results provide causal evidence that natural, object-associated signals in genetically-distinct but interacting hypothalamic neurons differentially control whether the brain forms object memories.
]]></description>
<dc:creator>Kosse, C.</dc:creator>
<dc:creator>Burdakov, D.</dc:creator>
<dc:date>2019-04-11</dc:date>
<dc:identifier>doi:10.1101/603936</dc:identifier>
<dc:title><![CDATA[Natural hypothalamic circuit dynamics underlying object memorization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/610741v1?rss=1">
<title>
<![CDATA[
nf-core: Community curated bioinformatics pipelines 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/610741v1?rss=1"
</link>
<description><![CDATA[
The standardization, portability, and reproducibility of analysis pipelines is a renowned problem within the bioinformatics community. Most pipelines are designed for execution on-premise, and the associated software dependencies are tightly coupled with the local compute environment. This leads to poor pipeline portability and reproducibility of the ensuing results - both of which are fundamental requirements for the validation of scientific findings. Here, we introduce nf-core: a framework that provides a community-driven, peer-reviewed platform for the development of best practice analysis pipelines written in Nextflow. Key obstacles in pipeline development such as portability, reproducibility, scalability and unified parallelism are inherently addressed by all nf-core pipelines. We are also continually developing a suite of tools that assist in the creation and development of both new and existing pipelines. Our primary goal is to provide a platform for high-quality, reproducible bioinformatics pipelines that can be utilized across various institutions and research facilities.
]]></description>
<dc:creator>Ewels, P.</dc:creator>
<dc:creator>Peltzer, A.</dc:creator>
<dc:creator>Fillinger, S.</dc:creator>
<dc:creator>Alneberg, J.</dc:creator>
<dc:creator>Patel, H.</dc:creator>
<dc:creator>Wilm, A.</dc:creator>
<dc:creator>Garcia, M.</dc:creator>
<dc:creator>Di Tommaso, P.</dc:creator>
<dc:creator>Nahnsen, S.</dc:creator>
<dc:date>2019-04-16</dc:date>
<dc:identifier>doi:10.1101/610741</dc:identifier>
<dc:title><![CDATA[nf-core: Community curated bioinformatics pipelines]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/611400v1?rss=1">
<title>
<![CDATA[
Epigenetic regulation of unique genes and repetitive elements by the KRAB zinc finger protein ZFP57 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/611400v1?rss=1"
</link>
<description><![CDATA[
BackgroundKRAB-zinc finger proteins (KZFPs) represent one of the largest families of DNA binding proteins in vertebrate genomes and appear to have evolved to silence transposable elements (TEs) including endogenous retroviruses through sequence-specific targeting of repressive chromatin states. ZFP57 is required to maintain the post-fertilization DNA methylation memory of parental-origin at genomic imprints along with ZFP445 which is specific for imprints. However, ZFP57 has multiple methylated genomic targets. Here we conduct RNA-seq and ChIP-seq analyses in normal and ZFP57 mutant mouse ES cells to understand the relative importance of ZFP57 at unique and repetitive regions of the genome.nnResultsOver 80% of ZFP57 targets are TEs, however, ZFP57 is not essential for their repression. The remaining targets lie within unique imprinted and non-imprinted sequences. Though loss of ZFP57 influences imprinted genes as expected, the majority of unique gene targets lose H3K9me3 with little effect on DNA methylation and very few exhibiting alterations in expression. Comparison with DNA methyltransferase-deleted ES cells (TKO) identifies remarkably similar losses of H3K9me3 and changes in expression, defining regions where H3K9me3 is secondary to DNA methylation. We show that ZFP57 is the principal methylation-sensitive KZFP recruiting KAP1 and H3K9me3 in ES cells. Finally, like imprints, other unique targets of ZFP57 are enriched for germline-derived DNA methylation including oocyte-specific methylation that is resistant to post-fertilisation epigenetic reprogramming.nnConclusionOur analyses suggest the evolution of a rare DNA methylation-sensitive KZFP that is not essential for repeat silencing, but whose primary function is to maintain DNA methylation and repressive histone marks at germline derived imprinting control regions.
]]></description>
<dc:creator>Shi, H.</dc:creator>
<dc:creator>Strogantsev, R.</dc:creator>
<dc:creator>Takahashi, N.</dc:creator>
<dc:creator>Kazachenka, A.</dc:creator>
<dc:creator>Lorincz, M.</dc:creator>
<dc:creator>Hemberger, M.</dc:creator>
<dc:creator>Ferguson-Smith, A.</dc:creator>
<dc:date>2019-04-18</dc:date>
<dc:identifier>doi:10.1101/611400</dc:identifier>
<dc:title><![CDATA[Epigenetic regulation of unique genes and repetitive elements by the KRAB zinc finger protein ZFP57]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/620096v1?rss=1">
<title>
<![CDATA[
Rapid sensory integration in orexin neurons governs probability of future movements 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/620096v1?rss=1"
</link>
<description><![CDATA[
Internally and externally triggered movement is crucial for survival and is controlled by multiple interconnected neuronal populations spanning many brain areas. Lateral hypothalamic orexin/hypocretin neurons are thought to play a slow, modulatory part in this scheme through their key role in promoting metabolism and wakefulness. However, it is unknown whether orexin/hypocretin neurons participate in rapid neural processing, leading to immediate movement. Furthermore, their role in sensorimotor transformations remains unknown. Here we use cellular-resolution Ca2+ imaging and optogenetics to show that orexin/hypocretin cells are instantaneous regulators of self-generated and sensory-evoked movement. They are activated before and during movement, preventing this activation reduces self-generated locomotion, and optogenetic mimicry of this transient activation rapidly initiates locomotion. We find that the same orexin neurons whose activity predicts movement initiation are rapidly controllable by external sensory stimuli, and silencing orexin cells during sensation prevents normal motor performance. These findings place orexin neurons in a physiological position of unexpectedly rapid and strong sensorimotor control.
]]></description>
<dc:creator>Karnani, M. M.</dc:creator>
<dc:creator>Schöne, C.</dc:creator>
<dc:creator>Bracey, E. F.</dc:creator>
<dc:creator>Gonzalez, J. A.</dc:creator>
<dc:creator>Viskaitis, P.</dc:creator>
<dc:creator>Adamantidis, A.</dc:creator>
<dc:creator>Burdakov, D.</dc:creator>
<dc:date>2019-04-26</dc:date>
<dc:identifier>doi:10.1101/620096</dc:identifier>
<dc:title><![CDATA[Rapid sensory integration in orexin neurons governs probability of future movements]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/644849v1?rss=1">
<title>
<![CDATA[
Altered Hippocampal-Prefrontal Neural Dynamics in Mouse Models of Down Syndrome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/644849v1?rss=1"
</link>
<description><![CDATA[
Altered neural dynamics in medial prefrontal cortex (mPFC) and hippocampus may contribute to cognitive impairments in the complex chromosomal disorder, Down Syndrome (DS). Here, we demonstrate non-overlapping behavioural differences associated with distinct abnormalities in hippocampal and mPFC electrophysiology during a canonical spatial memory task in three partially trisomic mouse models of DS (Dp1Tyb, Dp10Yey, Dp17Yey) that together cover all regions of homology with human chromosome 21 (Hsa21). Dp1Tyb mice showed slower decision-making (unrelated to the gene dose of DYRK1A, which has been implicated in DS cognitive dysfunction) and altered theta dynamics (reduced frequency, increased hippocampal-mPFC coherence, increased modulation of hippocampal high gamma); Dp10Yey mice showed impaired alternation performance and reduced theta modulation of hippocampal low gamma; while Dp17Yey mice were no different from wildtype mice. These results link specific hippocampal and mPFC circuit dysfunctions to cognitive deficits in DS models and, importantly, map them to discrete regions of Hsa21.
]]></description>
<dc:creator>Chang, P.</dc:creator>
<dc:creator>Bush, D.</dc:creator>
<dc:creator>Schorge, S.</dc:creator>
<dc:creator>Good, M.</dc:creator>
<dc:creator>Canonica, T.</dc:creator>
<dc:creator>Shing, N.</dc:creator>
<dc:creator>Noy, S.</dc:creator>
<dc:creator>Wiseman, F. K.</dc:creator>
<dc:creator>Burgess, N.</dc:creator>
<dc:creator>Tybulewicz, V. L.</dc:creator>
<dc:creator>Walker, M. C.</dc:creator>
<dc:creator>Fisher, E. M.</dc:creator>
<dc:date>2019-05-23</dc:date>
<dc:identifier>doi:10.1101/644849</dc:identifier>
<dc:title><![CDATA[Altered Hippocampal-Prefrontal Neural Dynamics in Mouse Models of Down Syndrome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/653105v1?rss=1">
<title>
<![CDATA[
The CAFA challenge reports improved protein function prediction and new functional annotations for hundreds of genes through experimental screens 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/653105v1?rss=1"
</link>
<description><![CDATA[
The Critical Assessment of Functional Annotation (CAFA) is an ongoing, global, community-driven effort to evaluate and improve the computational annotation of protein function. Here we report on the results of the third CAFA challenge, CAFA3, that featured an expanded analysis over the previous CAFA rounds, both in terms of volume of data analyzed and the types of analysis performed. In a novel and major new development, computational predictions and assessment goals drove some of the experimental assays, resulting in new functional annotations for more than 1000 genes. Specifically, we performed experimental whole-genome mutation screening in Candida albicans and Pseudomonas aureginosa genomes, which provided us with genome-wide experimental data for genes associated with biofilm formation and motility (P. aureginosa only). We further performed targeted assays on selected genes in Drosophila melanogaster, which we suspected of being involved in long-term memory. We conclude that, while predictions of the molecular function and biological process annotations have slightly improved over time, those of the cellular component have not. Term-centric prediction of experimental annotations remains equally challenging; although the performance of the top methods is significantly better than expectations set by baseline methods in C. albicans and D. melanogaster, it leaves considerable room and need for improvement. We finally report that the CAFA community now involves a broad range of participants with expertise in bioinformatics, biological experimentation, biocuration, and bioontologies, working together to improve functional annotation, computational function prediction, and our ability to manage big data in the era of large experimental screens.
]]></description>
<dc:creator>Zhou, N.</dc:creator>
<dc:creator>Jiang, Y.</dc:creator>
<dc:creator>Bergquist, T. R.</dc:creator>
<dc:creator>Lee, A. J.</dc:creator>
<dc:creator>Kacsoh, B. Z.</dc:creator>
<dc:creator>Crocker, A. W.</dc:creator>
<dc:creator>Lewis, K. A.</dc:creator>
<dc:creator>Georghiou, G.</dc:creator>
<dc:creator>Nguyen, H. N.</dc:creator>
<dc:creator>Hamid, M. N.</dc:creator>
<dc:creator>Davis, L.</dc:creator>
<dc:creator>The Critical Assessment of Function Annotation,</dc:creator>
<dc:creator>Rost, B.</dc:creator>
<dc:creator>Brenner, S. E.</dc:creator>
<dc:creator>Orengo, C. A.</dc:creator>
<dc:creator>Jeffery, C. J.</dc:creator>
<dc:creator>Bosco, G. D.</dc:creator>
<dc:creator>Hogan, D. A.</dc:creator>
<dc:creator>Martin, M. J.</dc:creator>
<dc:creator>O'Donovan, C.</dc:creator>
<dc:creator>Mooney, S. D.</dc:creator>
<dc:creator>Greene, C. S.</dc:creator>
<dc:creator>Radivojac, P.</dc:creator>
<dc:creator>Friedberg, I.</dc:creator>
<dc:date>2019-05-29</dc:date>
<dc:identifier>doi:10.1101/653105</dc:identifier>
<dc:title><![CDATA[The CAFA challenge reports improved protein function prediction and new functional annotations for hundreds of genes through experimental screens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/656793v1?rss=1">
<title>
<![CDATA[
ScanningSWATH enables ultra-fast proteomics using high-flow chromatography and minute-scale gradients 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/656793v1?rss=1"
</link>
<description><![CDATA[
Bridging genotype to phenotype, the proteome has increasingly become of major importance to generate large, longitudinal sample series for data-driven biology and personalized medicine. Major improvements in laboratory automation, chromatography and software have increased the scale and precision of proteomics. So far missing are however mass spectrometric acquisition techniques that could deal with very fast chromatographic gradients. Here we present scanning SWATH, a data-independent acquisition (DIA) method, in which the DIA-typical stepwise windowed acquisition is replaced by a continuous movement of the precursor isolation window. Scanning SWATH accelerates the duty cycles to a few hundreds of milliseconds, and enables precursor mass assignment to the MS2 fragment traces for improving true positive precursor identification in fast proteome experiments. In combination with 800 {micro}L/min high-flow chromatography, we report the quantification of 270 precursors per second, increasing the precursor identifications by 70% or more compared to previous methods. Scanning SWATH quantified 1,410 Human protein groups in conjunction with chromatographic gradients as fast as 30 seconds, 2,250 with 60-second gradients, and 4,586 in conjunction with 5-minute gradients. At high quantitative precision, our method hence increases the proteomic throughput to hundreds of samples per day per mass spectrometer. Scanning SWATH hence enables a broad range of new proteomic applications that depend on large numbers of cheap yet quantification precise proteomes.
]]></description>
<dc:creator>Messner, C.</dc:creator>
<dc:creator>Demichev, V.</dc:creator>
<dc:creator>Bloomfield, N.</dc:creator>
<dc:creator>Ivosev, G.</dc:creator>
<dc:creator>Wasim, F.</dc:creator>
<dc:creator>Zelezniak, A.</dc:creator>
<dc:creator>Lilley, K.</dc:creator>
<dc:creator>Tate, S.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:date>2019-05-31</dc:date>
<dc:identifier>doi:10.1101/656793</dc:identifier>
<dc:title><![CDATA[ScanningSWATH enables ultra-fast proteomics using high-flow chromatography and minute-scale gradients]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/659896v1?rss=1">
<title>
<![CDATA[
Single-Cell Signalling Analysis of Heterocellular Organoids 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/659896v1?rss=1"
</link>
<description><![CDATA[
Organoids are powerful biomimetic tissue models. Despite their widespread adoption, methods to analyse cell-type specific post-translational modification (PTM) signalling networks in organoids are absent. Here we report multivariate single-cell analysis of cell-type specific signalling networks in organoids and organoid co-cultures. Simultaneous measurement of 28 PTMs in >1 million single small intestinal organoid cells by mass cytometry reveals cell-type and cell-state specific signalling networks in stem, Paneth, enteroendocrine, tuft, goblet cells, and enterocytes. Integrating single-cell PTM analysis with Thiol-reactive Organoid Barcoding in situ (TOBis) enables high-throughput comparison of signalling networks between organoid cultures. Multivariate cell-type specific PTM analysis of colorectal cancer tumour microenvironment organoids reveals that shApc, KrasG12D, and Trp53R172H cell-autonomously mimic signalling states normally induced by stromal fibroblasts and macrophages. These results demonstrate how standard mass cytometry workflows can be modified to perform high-throughput multivariate cell-type specific signalling analysis of healthy and cancerous organoids.
]]></description>
<dc:creator>Qin, X.</dc:creator>
<dc:creator>Sufi, J.</dc:creator>
<dc:creator>Vlckova, P.</dc:creator>
<dc:creator>Kyriakidou, P.</dc:creator>
<dc:creator>Acton, S. E.</dc:creator>
<dc:creator>Li, V. S. W.</dc:creator>
<dc:creator>Nitz, M.</dc:creator>
<dc:creator>Tape, C. J.</dc:creator>
<dc:date>2019-06-06</dc:date>
<dc:identifier>doi:10.1101/659896</dc:identifier>
<dc:title><![CDATA[Single-Cell Signalling Analysis of Heterocellular Organoids]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/663336v1?rss=1">
<title>
<![CDATA[
Retroviral integration into nucleosomes throughDNA looping and sliding along the histone octamer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/663336v1?rss=1"
</link>
<description><![CDATA[
Retroviral integrase can efficiently utilise nucleosomes for insertion of the reverse-transcribed viral DNA. In face of the structural constraints imposed by the nucleosomal structure, integrase gains access to the scissile phosphodiester bonds by lifting DNA off the histone octamer at the site of integration. To clarify the mechanism of DNA looping by integrase, we determined a 3.9 [A] resolution structure of the prototype foamy virus intasome engaged with a nucleosome core particle. The structural data along with complementary single-molecule Forster resonance energy transfer measurements reveal twisting and sliding of the nucleosomal DNA arm proximal to the integration site. Sliding the nucleosomal DNA by approximately two base pairs along the histone octamer accommodates the necessary DNA lifting from the histone H2A-H2B subunits to allow engagement with the intasome. Thus, retroviral integration into nucleosomes involves the looping-and-sliding mechanism for nucleosomal DNA repositioning, bearing unexpected similarities with chromatin remodelers.
]]></description>
<dc:creator>Wilson, M. D.</dc:creator>
<dc:creator>Renault, L.</dc:creator>
<dc:creator>Maskell, D. P.</dc:creator>
<dc:creator>Ghoneim, M.</dc:creator>
<dc:creator>Pye, V. E.</dc:creator>
<dc:creator>Nans, A.</dc:creator>
<dc:creator>Rueda, D. S.</dc:creator>
<dc:creator>Cherepanov, P.</dc:creator>
<dc:creator>Costa, A.</dc:creator>
<dc:date>2019-06-07</dc:date>
<dc:identifier>doi:10.1101/663336</dc:identifier>
<dc:title><![CDATA[Retroviral integration into nucleosomes throughDNA looping and sliding along the histone octamer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/665836v1?rss=1">
<title>
<![CDATA[
Plasmodium Kinesin-8X associates with mitotic spindles and is essential for oocyst development during parasite proliferation and transmission 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/665836v1?rss=1"
</link>
<description><![CDATA[
Kinesin-8 proteins are microtubule motors that are often involved in regulation of mitotic spindle length and chromosome alignment. They move towards the ends of spindle microtubules and regulate the dynamics of these ends due, at least in some species, to their microtubule depolymerization activity. Plasmodium spp. exhibit an atypical endomitotic cell division in which chromosome condensation and spindle dynamics are not well understood in the different proliferative stages. Genome-wide homology analysis of Plasmodium spp. revealed the presence of two Kinesin-8 motor proteins (Kinesin-8X and Kinesin-8B). Here we have studied the biochemical properties of Kinesin-8X and its role in parasite proliferation. In vitro, Kinesin-8X showed motile and depolymerization activities like other Kinesin-8 motors. To understand its role in cell division, we have used protein tagging and live cell imaging to define the location of Plasmodium Kinesin-8X during all proliferative stages of the P berghei life cycle. Furthermore, we have used gene targeting to analyse the function of Kinesin-8X. The results reveal a spatio-temporal involvement of Kinesin-8X in spindle dynamics and its association with both mitotic and meiotic spindles and the putative microtubule organising centre (MTOC). Deletion of the Kinesin-8X gene showed that this protein is required for endomitotic division during oocyst development and is therefore necessary for parasite replication within the mosquito gut, and for transmission to the vertebrate host. Consistently, transcriptome analysis of{Delta} kinesin-8X parasites reveals modulated expression of genes involved mainly in microtubule-based processes, chromosome organisation and the regulation of gene expression supporting a role in cell division.nnAuthor SummaryKinesins are microtubule-based motors that play key roles in intracellular transport, cell division and motility. Members of the Kinesin-8 family contribute to chromosome alignment during cell division in many eukaryotes. However, the roles of kinesins in the atypical cell division in Plasmodium, the causative agent of malaria, is not known. In contrast to many other eukaryotes, Plasmodium proliferates by endomitosis, in which genome replication and division occur within a nucleus bounded by a persistent nuclear envelope. We show that the Plasmodium genome encodes only nine kinesins and we further investigate the role of Kinesin-8X throughout the Plasmodium life cycle using biochemical and gene targeting approaches. We show that Plasmodium Kinesin-8X has microtubule-based motility and depolymerization activity. We also show that Kinesin-8X is probably localized on putative MTOCs and spindles during cell division in most of the stages of P. berghei life cycle. By gene deletion we demonstrate that Kinesin-8X is essential for normal oocyst development and sporozoite formation. Genome-wide RNA analysis of{Delta} kinesin-8X parasites reveals modulated expression of genes involved in microtubule-based processes. Overall, the data suggest that Kinesin-8X is a molecular motor that plays essential roles during endomitosis in oocyst development in the mosquito, contributing to parasite transmission.
]]></description>
<dc:creator>Zeeshan, M.</dc:creator>
<dc:creator>Shilliday, F.</dc:creator>
<dc:creator>Liu, T.</dc:creator>
<dc:creator>Abel, S.</dc:creator>
<dc:creator>Mourier, T.</dc:creator>
<dc:creator>Ferguson, D. J. P.</dc:creator>
<dc:creator>Rea, E.</dc:creator>
<dc:creator>Stanway, R. R.</dc:creator>
<dc:creator>Roques, M.</dc:creator>
<dc:creator>Williams, D.</dc:creator>
<dc:creator>Daniel, E.</dc:creator>
<dc:creator>Brady, D.</dc:creator>
<dc:creator>Roberts, A. J.</dc:creator>
<dc:creator>Sinden, R. E.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:creator>Pain, A.</dc:creator>
<dc:creator>Le Roch, K. G.</dc:creator>
<dc:creator>Moores, C. A.</dc:creator>
<dc:creator>Tewari, R.</dc:creator>
<dc:date>2019-06-10</dc:date>
<dc:identifier>doi:10.1101/665836</dc:identifier>
<dc:title><![CDATA[Plasmodium Kinesin-8X associates with mitotic spindles and is essential for oocyst development during parasite proliferation and transmission]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/671578v1?rss=1">
<title>
<![CDATA[
Systematic analysis of Plasmodium myosins reveals differential expression, localization and function in invasive and proliferative parasite stages 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/671578v1?rss=1"
</link>
<description><![CDATA[
The myosin superfamily comprises of actin-dependent eukaryotic molecular motors important in a variety of cellular functions. Although well studied in many systems, knowledge of their functions in Plasmodium, the causative agent of malaria, is restricted. Previously, six myosins were identified in this genus, including three Class XIV myosins found only in Apicomplexa and some Ciliates. The well characterised MyoA, is a class XIV myosin essential for gliding motility and invasion. Here, we characterize all other Plasmodium myosins throughout the parasite life cycle and show that they have very diverse patterns of expression and cellular location. MyoB and MyoE, the other two Class XIV myosins, are expressed in all invasive stages, with apical and basal locations, respectively. Gene deletion revealed that MyoE is involved in sporozoite motility, MyoF and MyoK are likely essential in the asexual blood stages, and MyoJ and MyoB are not essential. Both MyoB and its essential light chain (MCL-B) are localised at the apical end of ookinetes but expressed at completely different time points. This work provides a better understanding of the role of actomyosin motors in Apicomplexan parasites, particularly in the motile and invasive stages of Plasmodium during sexual and asexual development within the mosquito.
]]></description>
<dc:creator>Wall, R. J.</dc:creator>
<dc:creator>ZEESHAN, M.</dc:creator>
<dc:creator>Katris, N. J.</dc:creator>
<dc:creator>Limenitakis, R.</dc:creator>
<dc:creator>Rea, E.</dc:creator>
<dc:creator>Stock, J.</dc:creator>
<dc:creator>Brady, D.</dc:creator>
<dc:creator>Waller, R. F.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:creator>Tewari, R.</dc:creator>
<dc:date>2019-06-14</dc:date>
<dc:identifier>doi:10.1101/671578</dc:identifier>
<dc:title><![CDATA[Systematic analysis of Plasmodium myosins reveals differential expression, localization and function in invasive and proliferative parasite stages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/672576v1?rss=1">
<title>
<![CDATA[
Inherited duplications of PPP2R3B promote naevi and melanoma via a novel C21orf91-driven proliferative phenotype 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/672576v1?rss=1"
</link>
<description><![CDATA[
The majority of the heredity of melanoma remains unexplained, however inherited copy number changes have not yet been systematically studied. The genetic environment is highly relevant to treatment stratification, and new gene discovery is therefore desirable. Using an unbiased whole genome screening approach for copy number we identify here a novel melanoma predisposing factor, familial duplications of gene PPP2R3B, encoding a regulatory unit of critical phosphatase PP2A. Significant correlation between expression of PPP2R3B in tumour tissue and survival in a large melanoma cohort was confirmed, and associated with a non-immunological expression profile. Mechanistically, construction and extensive characterization of a stable, inducible cellular model for PPP2R3B overexpression revealed induction of pigment cell switching towards proliferation and away from migration. Importantly, this was independent of the known microphthalmia-associated transcription factor (MITF)-controlled pigment cell phenotype switch, and was instead driven by uncharacterised gene C21orf91. Bioinformatic studies point to C21orf91as a novel target of MITF, and therefore a potential hub in the control of phenotype switching in melanoma. This study identifies novel germline copy number variants in PPP2R3B predisposing to melanocytic neoplasia, and uncovers a new potential therapeutic target C21orf91 in the control of pigment cell proliferation.
]]></description>
<dc:creator>Polubothu, S.</dc:creator>
<dc:creator>Al-Olabi, L.</dc:creator>
<dc:creator>Lionarons, D. A.</dc:creator>
<dc:creator>Harland, M.</dc:creator>
<dc:creator>Thomas, A.</dc:creator>
<dc:creator>Horswell, S.</dc:creator>
<dc:creator>Hunt, L.</dc:creator>
<dc:creator>Wlodarchak, N.</dc:creator>
<dc:creator>Aguilera, P.</dc:creator>
<dc:creator>Brand, S.</dc:creator>
<dc:creator>Bryant, D.</dc:creator>
<dc:creator>Beales, P.</dc:creator>
<dc:creator>Carrera, C.</dc:creator>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Elgar, G.</dc:creator>
<dc:creator>Harwood, C. A.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>Jenkins, D.</dc:creator>
<dc:creator>Larue, L.</dc:creator>
<dc:creator>Loughlin, S.</dc:creator>
<dc:creator>MacDonald, J.</dc:creator>
<dc:creator>Malvehy, J.</dc:creator>
<dc:creator>Barberan Martin, S.</dc:creator>
<dc:creator>Martins da Silva, V.</dc:creator>
<dc:creator>Molina, M.</dc:creator>
<dc:creator>Morrogh, D.</dc:creator>
<dc:creator>Moulding, D.</dc:creator>
<dc:creator>Nsengimana, J.</dc:creator>
<dc:creator>Pittman, A.</dc:creator>
<dc:creator>Puig-Butille, J.-A.</dc:creator>
<dc:creator>Parmar, K.</dc:creator>
<dc:creator>Sebire, N. J.</dc:creator>
<dc:creator>Scherer, S.</dc:creator>
<dc:creator>Stadnik, P.</dc:creator>
<dc:creator>Stanier, P.</dc:creator>
<dc:creator>Tell-Marti, G.</dc:creator>
<dc:creator>Waelchli, R.</dc:creator>
<dc:creator>Zarrei, M.</dc:creator>
<dc:creator>Zecchin, D.</dc:creator>
<dc:creator>Puig, S.</dc:creator>
<dc:creator>Bataille, V.</dc:creator>
<dc:creator>Xing, Y.</dc:creator>
<dc:creator>Healey, E.</dc:creator>
<dc:creator>Moore, G. E.</dc:creator>
<dc:date>2019-06-15</dc:date>
<dc:identifier>doi:10.1101/672576</dc:identifier>
<dc:title><![CDATA[Inherited duplications of PPP2R3B promote naevi and melanoma via a novel C21orf91-driven proliferative phenotype]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/674028v1?rss=1">
<title>
<![CDATA[
Plasmodium condensin core subunits (SMC2/SMC4) mediate atypical mitosis and are essential for parasite proliferation and transmission 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/674028v1?rss=1"
</link>
<description><![CDATA[
Condensin is a multi-subunit protein complex regulating chromosome condensation and segregation during cell division. In Plasmodium spp., the causative agent of malaria, cell division is atypical and the role of condensin is unclear. Here we examine the role of SMC2 and SMC4, the core subunits of condensin, during endomitosis in schizogony and endoreduplication in male gametogenesis. During early schizogony SMC2/SMC4 localize to a distinct focus, identified as the centromeres by NDC80 fluorescence and ChIP-seq analyses, but do not form condensin I or II complexes. In mature schizonts and during male gametogenesis, there is a diffuse SMC2/SMC4 distribution on chromosomes and in the nucleus, and both condensin I and II complexes form at these stages. Knockdown of smc2 and smc4 gene expression revealed essential roles in parasite proliferation and transmission. The condensin core subunits (SMC2/SMC4) form different complexes and may have distinct functions at various stages of the parasite life cycle.
]]></description>
<dc:creator>Pandey, R.</dc:creator>
<dc:creator>Abel, S.</dc:creator>
<dc:creator>Boucher, M.</dc:creator>
<dc:creator>Wall, R. J.</dc:creator>
<dc:creator>Zeeshan, M.</dc:creator>
<dc:creator>Rea, E.</dc:creator>
<dc:creator>Freville, A.</dc:creator>
<dc:creator>Lu, X. M.</dc:creator>
<dc:creator>Brady, D.</dc:creator>
<dc:creator>Daniel, E.</dc:creator>
<dc:creator>Stanway, R. R.</dc:creator>
<dc:creator>Wheatley, S.</dc:creator>
<dc:creator>Batugedara, G.</dc:creator>
<dc:creator>Hollin, T.</dc:creator>
<dc:creator>Bottrill, A. R.</dc:creator>
<dc:creator>Gupta, D.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:creator>Le Roch, K. G.</dc:creator>
<dc:creator>Tewari, R.</dc:creator>
<dc:date>2019-06-17</dc:date>
<dc:identifier>doi:10.1101/674028</dc:identifier>
<dc:title><![CDATA[Plasmodium condensin core subunits (SMC2/SMC4) mediate atypical mitosis and are essential for parasite proliferation and transmission]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/680561v1?rss=1">
<title>
<![CDATA[
PEGylated surfaces for the study of DNA-protein interactions by atomic force microscopy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/680561v1?rss=1"
</link>
<description><![CDATA[
DNA-protein interactions are vital to cellular function, with key roles in the regulation of gene expression and genome maintenance. Atomic force microscopy (AFM) offers the ability to visualize DNA-protein interactions at nanometre resolution in near-physiological buffers, but it requires that the DNA be adhered to the surface of a solid substrate. This presents a problem when working at biologically relevant protein concentrations, where protein may be present at large excess in solution; much of the biophysically relevant information can therefore be occluded by non-specific protein binding to the underlying substrate. Here we explore the use of PLLx-b-PEGy block copolymers to achieve selective adsorption of DNA on a mica surface. Through varying both the number of lysine and ethylene glycol residues in the block copolymers, we show selective adsorption of DNA on mica that is functionalized with a PLL10-b-PEG113 / PLL1000-2000 mixture as viewed by AFM imaging in a solution containing high concentrations of streptavidin. We show that this selective adsorption extends to DNA-protein complexes, through the use of biotinylated DNA and streptavidin, and demonstrate that DNA-bound streptavidin can be unambiguously distinguished by in-liquid AFM in spite of an excess of unbound streptavidin in solution.
]]></description>
<dc:creator>Akpinar, B.</dc:creator>
<dc:creator>Bell, N.</dc:creator>
<dc:creator>Pyne, A.</dc:creator>
<dc:creator>Hoogenboom, B.</dc:creator>
<dc:date>2019-06-24</dc:date>
<dc:identifier>doi:10.1101/680561</dc:identifier>
<dc:title><![CDATA[PEGylated surfaces for the study of DNA-protein interactions by atomic force microscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/682450v1?rss=1">
<title>
<![CDATA[
Broadly inhibiting anti-neuraminidase monoclonal antibodies induced by trivalent influenza vaccine and H7N9 infection in humans 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/682450v1?rss=1"
</link>
<description><![CDATA[
The majority of antibodies induced by influenza neuraminidase (NA), like those against hemagglutinin (HA), are relatively specific to viruses isolated within a limited time-window as seen in serological studies and the analysis of many murine monoclonal antibodies. We report three broadly reactive human monoclonal antibodies (mAbs) targeting N1 NA. Two were isolated from a young adult vaccinated with trivalent influenza vaccine (TIV), which inhibited N1 NA from viruses isolated from human over a period of a hundred years. The third antibody isolated from a child with acute mild H7N9 infection inhibited both group 1 N1 and group 2 N9 NAs. In addition, the antibodies cross-inhibited the N1 NAs of highly pathogenic avian H5N1 influenza viruses. These antibodies are protective in prophylaxis against seasonal H1N1 viruses in mice. This study demonstrates that human antibodies to N1 NA with exceptional cross-reactivity can be recalled by vaccination and highlights the importance of standardizing the NA antigen in seasonal vaccines to offer optimal protection.nnImportanceAntibodies to the influenza NA can provide protection against influenza disease. Analysis of human antibodies to NA lags behind that for HA. We show that human monoclonal antibodies against NA induced by vaccination and infection can be very broadly reactive and able to inhibit a wide spectrum of N1 NAs between 1918 and 2018. This suggests that antibodies to NA may be a useful therapy, and that efficacy of influenza vaccines could be enhanced by ensuring appropriate content of NA antigen.nnHighlights of the paperO_LIAntibodies that inhibit influenza viruses with N1 neuraminidase (NA), with broad reactivity for viruses isolated between 1918-2018, can be isolated from human recipients of seasonal influenza vaccinenC_LIO_LIAntibodies targeting N1 NA of human seasonal H1N1 viruses can cross-react with a variety of avian N1 neuraminidasesnC_LIO_LIAcute H7N9 infection can recall memory B cells to N1 NA and elicit cross-reactive antibodies to the group 1 N1 and group 2 N9 NAsnC_LIO_LIAntibodies to N1 NA with this broad reactivity protect against lethal virus challengenC_LI
]]></description>
<dc:creator>Rijal, P.</dc:creator>
<dc:creator>Wang, B. B.</dc:creator>
<dc:creator>Tan, T. K.</dc:creator>
<dc:creator>Schimanski, L.</dc:creator>
<dc:creator>Janesch, P.</dc:creator>
<dc:creator>Dong, T.</dc:creator>
<dc:creator>McCauley, J. M.</dc:creator>
<dc:creator>Daniels, R. S.</dc:creator>
<dc:creator>Townsend, A.</dc:creator>
<dc:creator>Huang, K.-Y. A.</dc:creator>
<dc:date>2019-06-26</dc:date>
<dc:identifier>doi:10.1101/682450</dc:identifier>
<dc:title><![CDATA[Broadly inhibiting anti-neuraminidase monoclonal antibodies induced by trivalent influenza vaccine and H7N9 infection in humans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/681924v1?rss=1">
<title>
<![CDATA[
RADICL-seq identifies general and cell type-specific principles of genome-wide RNA-chromatin interactions 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/681924v1?rss=1"
</link>
<description><![CDATA[
Mammalian genomes encode tens of thousands of noncoding RNAs. Most noncoding transcripts exhibit nuclear localization and several have been shown to play a role in the regulation of gene expression and chromatin remodelling. To investigate the function of such RNAs, methods to massively map the genomic interacting sites of multiple transcripts have been developed. However, they still present some limitations. Here, we introduce RNA And DNA Interacting Complexes Ligated and sequenced (RADICL-seq), a technology that maps genome-wide RNA-chromatin interactions in intact nuclei. RADICL-seq is a proximity ligation-based methodology that reduces the bias for nascent transcription, while increasing genomic coverage and unique mapping rate efficiency compared to existing methods. RADICL-seq identifies distinct patterns of genome occupancy for different classes of transcripts as well as cell type-specific RNA-chromatin interactions, and emphasizes the role of transcription in the establishment of chromatin structure.
]]></description>
<dc:creator>Bonetti, A.</dc:creator>
<dc:creator>Agostini, F.</dc:creator>
<dc:creator>Suzuki, A. M.</dc:creator>
<dc:creator>Hashimoto, K.</dc:creator>
<dc:creator>Pascarella, G.</dc:creator>
<dc:creator>Gimenez, J.</dc:creator>
<dc:creator>Roos, L.</dc:creator>
<dc:creator>Nash, A. J.</dc:creator>
<dc:creator>Ghilotti, M.</dc:creator>
<dc:creator>Cameron, C. J.</dc:creator>
<dc:creator>Valentine, M.</dc:creator>
<dc:creator>Medvedeva, Y. A.</dc:creator>
<dc:creator>Noguchi, S.</dc:creator>
<dc:creator>Agirre, E.</dc:creator>
<dc:creator>Kashi, K.</dc:creator>
<dc:creator>Samudyata, S.</dc:creator>
<dc:creator>Luginbuehl, J.</dc:creator>
<dc:creator>Cazzoli, R.</dc:creator>
<dc:creator>Agrawal, S.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:creator>Blanchette, M.</dc:creator>
<dc:creator>Kasukawa, T.</dc:creator>
<dc:creator>De Hoon, M.</dc:creator>
<dc:creator>Arner, E.</dc:creator>
<dc:creator>Lenhard, B.</dc:creator>
<dc:creator>Plessy, C.</dc:creator>
<dc:creator>Castelo-Branco, G.</dc:creator>
<dc:creator>Orlando, V.</dc:creator>
<dc:creator>Carninci, P.</dc:creator>
<dc:date>2019-06-27</dc:date>
<dc:identifier>doi:10.1101/681924</dc:identifier>
<dc:title><![CDATA[RADICL-seq identifies general and cell type-specific principles of genome-wide RNA-chromatin interactions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/674986v1?rss=1">
<title>
<![CDATA[
Insights into human genetic variation and population history from 929 diverse genomes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/674986v1?rss=1"
</link>
<description><![CDATA[
Genome sequences from diverse human groups are needed to understand the structure of genetic variation in our species and the history of, and relationships between, different populations. We present 929 high-coverage genome sequences from 54 diverse human populations, 26 of which are physically phased using linked-read sequencing. Analyses of these genomes reveal an excess of previously undocumented private genetic variation in southern and central Africa and in Oceania and the Americas, but an absence of fixed, private variants between major geographical regions. We also find deep and gradual population separations within Africa, contrasting population size histories between hunter-gatherer and agriculturalist groups in the last 10,000 years, a potentially major population growth episode after the peopling of the Americas, and a contrast between single Neanderthal but multiple Denisovan source populations contributing to present-day human populations. We also demonstrate benefits to the study of population relationships of genome sequences over ascertained array genotypes. These genome sequences are freely available as a resource with no access or analysis restrictions.
]]></description>
<dc:creator>Bergström, A.</dc:creator>
<dc:creator>McCarthy, S. A.</dc:creator>
<dc:creator>Hui, R.</dc:creator>
<dc:creator>Almarri, M. A.</dc:creator>
<dc:creator>Ayub, Q.</dc:creator>
<dc:creator>Danecek, P.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Felkel, S.</dc:creator>
<dc:creator>Hallast, P.</dc:creator>
<dc:creator>Kamm, J.</dc:creator>
<dc:creator>Blanche, H.</dc:creator>
<dc:creator>Deleuze, J.-F.</dc:creator>
<dc:creator>Cann, H.</dc:creator>
<dc:creator>Mallick, S.</dc:creator>
<dc:creator>Reich, D.</dc:creator>
<dc:creator>Sandhu, M. S.</dc:creator>
<dc:creator>Skoglund, P.</dc:creator>
<dc:creator>Scally, A.</dc:creator>
<dc:creator>Xue, Y.</dc:creator>
<dc:creator>Durbin, R.</dc:creator>
<dc:creator>Tyler-Smith, C.</dc:creator>
<dc:date>2019-06-27</dc:date>
<dc:identifier>doi:10.1101/674986</dc:identifier>
<dc:title><![CDATA[Insights into human genetic variation and population history from 929 diverse genomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/683631v1?rss=1">
<title>
<![CDATA[
Actomyosin regulation by Eph receptor signaling couples boundary cell formation to border sharpness 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/683631v1?rss=1"
</link>
<description><![CDATA[
The segregation of cells with distinct regional identity underlies formation of a sharp border, which in some tissues serves to organise a boundary signaling centre. It is unclear whether or how border sharpness is coordinated with induction of boundary-specific gene expression. We show that forward signaling of EphA4 is required for border sharpening and induction of boundary cells in the zebrafish hindbrain, which we find both require kinase-dependent signaling, with a lesser input of PDZ domain-dependent signaling. We find that boundary-specific gene expression is regulated by myosin II phosphorylation, which increases actomyosin contraction downstream of EphA4 signaling. Myosin phosphorylation leads to nuclear translocation of Taz, which together with Tead1a is required for boundary marker expression. Since actomyosin contraction maintains sharp borders, there is direct coupling of border sharpness to boundary cell induction that ensures correct organisation of signaling centres.
]]></description>
<dc:creator>Cayuso, J.</dc:creator>
<dc:creator>Xu, Q.</dc:creator>
<dc:creator>Addison, M.</dc:creator>
<dc:creator>Wilkinson, D.</dc:creator>
<dc:date>2019-06-27</dc:date>
<dc:identifier>doi:10.1101/683631</dc:identifier>
<dc:title><![CDATA[Actomyosin regulation by Eph receptor signaling couples boundary cell formation to border sharpness]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/686626v1?rss=1">
<title>
<![CDATA[
Mouse retinal cell behaviour in space and time using light sheet fluorescence microscopy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/686626v1?rss=1"
</link>
<description><![CDATA[
As the general population ages and the incidence of diabetes increases epidemically, more people are affected by eye diseases, such as retinopathies. It is therefore critical to improve imaging of eye disease mouse models. Here, we demonstrate that 1) rapid, quantitative 3D and 4D (time lapse) imaging of cellular and subcellular processes in the murine eye is feasible, with and without tissue clearing, using light-sheet fluorescent microscopy (LSFM) and 2) LSFM readily reveals new features of even well studied eye disease mouse models, such as the Oxygen-Induced Retinopathy (OIR) model. Through correlative LSFM-Confocal imaging we find that flat-mounting retinas for confocal microscopy significantly distorts tissue morphology. The minimized distortion with LSFM dramatically improved analysis of pathological vascular tufts in the OIR model revealing "knotted" morphologies, leading to a proposed new tuft nomenclature. Furthermore, live-imaging of OIR tuft formation revealed abnormal cell motility and altered filopodia dynamics. We conclude that quantitative 3D/4D LSFM imaging and analysis has the potential to advance our understanding of pathological processes in the eye, in particular neuro-vascular degenerative processes.
]]></description>
<dc:creator>Prahst, C.</dc:creator>
<dc:creator>Ashrafzadeh, P.</dc:creator>
<dc:creator>Harrington, K.</dc:creator>
<dc:creator>Venkatraman, L.</dc:creator>
<dc:creator>Richards, M.</dc:creator>
<dc:creator>Russo, A. M.</dc:creator>
<dc:creator>Cho, K.-S.</dc:creator>
<dc:creator>Chang, K.</dc:creator>
<dc:creator>Mead, T.</dc:creator>
<dc:creator>Chen, D. F.</dc:creator>
<dc:creator>Richardson, D.</dc:creator>
<dc:creator>Claesson-Welsh, L.</dc:creator>
<dc:creator>Franco, C.</dc:creator>
<dc:creator>Bentley, K.</dc:creator>
<dc:date>2019-06-28</dc:date>
<dc:identifier>doi:10.1101/686626</dc:identifier>
<dc:title><![CDATA[Mouse retinal cell behaviour in space and time using light sheet fluorescence microscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/686568v1?rss=1">
<title>
<![CDATA[
Kinesin-8B controls basal body function and flagellum formation and is key to malaria parasite transmission 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/686568v1?rss=1"
</link>
<description><![CDATA[
Eukaryotic flagella are conserved microtubule-based organelles that drive cell motility. Plasmodium, the causative agent of malaria, has a single flagellate stage: the male gamete in the mosquito. Three rounds of endomitotic division together with an unusual mode of flagellum assembly rapidly produce eight motile gametes. These processes are tightly coordinated but their regulation is poorly understood. To understand this important developmental stage, we studied the function and location of the microtubule-based motor kinesin-8B, using gene-targeting, electron microscopy and live cell imaging. Deletion of the kinesin-8B gene showed no effect on mitosis but disrupted 9+2 axoneme assembly and flagellum formation during male gamete development and also completely ablated parasite transmission. Live cell imaging showed that kinesin-8B-GFP did not colocalise with kinetochores in the nucleus but instead revealed dynamic, cytoplasmic localisation with the basal bodies and the assembling axoneme during flagellum formation. We thus uncovered an unexpected role for kinesin-8B in parasite flagellum formation that is vital for the parasite life cycle.
]]></description>
<dc:creator>Zeeshan, M.</dc:creator>
<dc:creator>Ferguson, D. J. P.</dc:creator>
<dc:creator>Abel, S.</dc:creator>
<dc:creator>Burrrell, A.</dc:creator>
<dc:creator>Rea, E.</dc:creator>
<dc:creator>Brady, D.</dc:creator>
<dc:creator>Daniel, E.</dc:creator>
<dc:creator>Delves, M.</dc:creator>
<dc:creator>Vaughan, S.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:creator>Le Roch, K. G.</dc:creator>
<dc:creator>Moores, C. A.</dc:creator>
<dc:creator>Tewari, R.</dc:creator>
<dc:date>2019-06-28</dc:date>
<dc:identifier>doi:10.1101/686568</dc:identifier>
<dc:title><![CDATA[Kinesin-8B controls basal body function and flagellum formation and is key to malaria parasite transmission]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/688010v1?rss=1">
<title>
<![CDATA[
High-throughput phenotyping reveals expansive genetic and structural underpinnings of immune variation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/688010v1?rss=1"
</link>
<description><![CDATA[
By developing a high-density murine immunophenotyping platform compatible with high-throughput genetic screening, we have established profound contributions of genetics and structure to immune variation. Specifically, high-throughput phenotyping of 530 knockout mouse lines identified 140 monogenic "hits" (>25%), most of which had never hitherto been implicated in immunology. Furthermore, they were conspicuously enriched in genes for which humans show poor tolerance to loss-of-function. The immunophenotyping platform also exposed dense correlation networks linking immune parameters with one another and with specific physiologic traits. By limiting the freedom of individual immune parameters, such linkages impose genetically regulated "immunological structures", whose integrity was found to be associated with immunocompetence. Hence, our findings provide an expanded genetic resource and structural perspective for understanding and monitoring immune variation in health and disease.
]]></description>
<dc:creator>Abeler-Dörner, L.</dc:creator>
<dc:creator>Laing, A.</dc:creator>
<dc:creator>Lorenc, A.</dc:creator>
<dc:creator>Ushakov, D. S.</dc:creator>
<dc:creator>Clare, S.</dc:creator>
<dc:creator>Speak, A.</dc:creator>
<dc:creator>Duque, M.</dc:creator>
<dc:creator>White, J. K.</dc:creator>
<dc:creator>Ramirez-Solis, R.</dc:creator>
<dc:creator>Saran, N.</dc:creator>
<dc:creator>Bull, K. R.</dc:creator>
<dc:creator>Moron, B.</dc:creator>
<dc:creator>Iwasaki, J.</dc:creator>
<dc:creator>Barton, P. R.</dc:creator>
<dc:creator>Caetano, S.</dc:creator>
<dc:creator>Hng, K. I.</dc:creator>
<dc:creator>Cambridge, E.</dc:creator>
<dc:creator>Forman, S.</dc:creator>
<dc:creator>Crockford, T. L.</dc:creator>
<dc:creator>Griffiths, M.</dc:creator>
<dc:creator>Kane, L.</dc:creator>
<dc:creator>Harcourt, K.</dc:creator>
<dc:creator>Brandt, C.</dc:creator>
<dc:creator>Notley, G.</dc:creator>
<dc:creator>Babalola, K. O.</dc:creator>
<dc:creator>Warren, J.</dc:creator>
<dc:creator>Mason, J. C.</dc:creator>
<dc:creator>Meeniga, A.</dc:creator>
<dc:creator>Karp, N. A.</dc:creator>
<dc:creator>Melvin, D.</dc:creator>
<dc:creator>Cawthorne, E.</dc:creator>
<dc:creator>Weinrick, B.</dc:creator>
<dc:creator>Rahim, A.</dc:creator>
<dc:creator>Drissler, S.</dc:creator>
<dc:creator>Meskas, J.</dc:creator>
<dc:creator>Yue, A.</dc:creator>
<dc:creator>Lux, M.</dc:creator>
<dc:creator>Song-Zhao, G.</dc:creator>
<dc:creator>Chan, A.</dc:creator>
<dc:creator>Ballesteros Reviriego, C.</dc:creator>
<dc:creator>Abeler, J.</dc:creator>
<dc:creator>Wilson, H.</dc:creator>
<dc:creator>Przemska-Kosicka, A.</dc:creator>
<dc:creator>Edmans, M.</dc:creator>
<dc:creator>Stre</dc:creator>
<dc:date>2019-07-02</dc:date>
<dc:identifier>doi:10.1101/688010</dc:identifier>
<dc:title><![CDATA[High-throughput phenotyping reveals expansive genetic and structural underpinnings of immune variation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/690172v1?rss=1">
<title>
<![CDATA[
Cyclin-dependent-like kinase 5 is required for pain signalling in both human neurons and mouse models 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/690172v1?rss=1"
</link>
<description><![CDATA[
Cyclin-dependent-like kinase 5 (Cdkl5) gene mutations lead to an X-linked disorder that is characterized by infantile epileptic encephalopathy, developmental delay and hypotonia. However, we found that a substantial percentage of these patients also report a previously unrecognised anamnestic deficiency in pain perception. Consistent with a role in nociception, we discovered that Cdkl5 is expressed selectively in nociceptive dorsal root ganglia (DRG) neurons in mice and in iPS-derived human nociceptors. CDKL5 deficient mice display defective epidermal innervation and conditional deletion of Cdkl5 in DRG sensory neurons significantly impairs nociception, phenocopying CDKL5 deficiency disorder in patients. Mechanistically, Cdkl5 interacts with CaMKII to control outgrowth as well as TRPV1-dependent signalling, which are disrupted in both Cdkl5 mutant murine DRG and human iPS-derived nociceptors. Together, these findings unveil a previously unrecognized role for Cdkl5 in nociception, proposing an original regulatory mechanism for pain perception with implications for future therapeutics in CDKL5 deficiency disorder.nnOne Sentence SummaryCyclin-dependent-like kinase 5 (Cdkl5) controls nociception in patients and murine models of Cdkl5 deficiency disorder via CaMKII-dependent mechanisms
]]></description>
<dc:creator>La Montanara, P.</dc:creator>
<dc:creator>Hervera Abad, A.</dc:creator>
<dc:creator>Baltussen, L.</dc:creator>
<dc:creator>Palmisano, I.</dc:creator>
<dc:creator>De Virgiliis, F.</dc:creator>
<dc:creator>Gao, Y.</dc:creator>
<dc:creator>Qasim, K.</dc:creator>
<dc:creator>Gorgoraptis, N.</dc:creator>
<dc:creator>Wong, K.</dc:creator>
<dc:creator>Downs, J.</dc:creator>
<dc:creator>Di Lazzaro, V.</dc:creator>
<dc:creator>Pizzorusso, T.</dc:creator>
<dc:creator>Ultanir, S.</dc:creator>
<dc:creator>Leonard, H.</dc:creator>
<dc:creator>Nagy, I.</dc:creator>
<dc:creator>Mazarakis, N.</dc:creator>
<dc:creator>Di Giovanni, S.</dc:creator>
<dc:date>2019-07-02</dc:date>
<dc:identifier>doi:10.1101/690172</dc:identifier>
<dc:title><![CDATA[Cyclin-dependent-like kinase 5 is required for pain signalling in both human neurons and mouse models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/692095v1?rss=1">
<title>
<![CDATA[
Greb1 is required for axial elongation and segmentation in vertebrate embryos. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/692095v1?rss=1"
</link>
<description><![CDATA[
During vertebrate embryonic development, the formation of axial structures is driven by a population of stem-like cells that reside in a region of the tailbud called the chordoneural hinge (CNH). We have compared the CNH transcriptome with those of surrounding tissues and shown that the CNH and tailbud mesoderm are transcriptionally similar, and distinct from the presomitic mesoderm. Amongst CNH-enriched genes are several that are required for axial elongation, including Wnt3a, Cdx2, Brachyury/T and Fgf8, and androgen/estrogen receptor nuclear signalling components such as Greb1. We show that the pattern and duration of tailbud Greb1 expression is conserved in mouse, zebrafish, and chicken embryos, and that Greb1 is required for axial elongation and somitogenesis in zebrafish embryos. The axial truncation phenotype of Greb1 morphant embryos is explained by much reduced expression of No tail (Ntl/Brachyury) which is required for axial progenitor maintenance. Posterior segmentation defects in the morphants (including misexpression of genes such as mespb, myoD and papC) appear to result, in part, from lost expression of the segmentation clock gene, her7.
]]></description>
<dc:creator>Prajapati, R. S.</dc:creator>
<dc:creator>Mitter, R.</dc:creator>
<dc:creator>Vezzaro, A.</dc:creator>
<dc:creator>Ish-Horowicz, D.</dc:creator>
<dc:date>2019-07-06</dc:date>
<dc:identifier>doi:10.1101/692095</dc:identifier>
<dc:title><![CDATA[Greb1 is required for axial elongation and segmentation in vertebrate embryos.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/697334v1?rss=1">
<title>
<![CDATA[
Perifornical Area Urocortin-3 Neurons Promote Infant-directed Neglect and Aggression 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/697334v1?rss=1"
</link>
<description><![CDATA[
While recent studies have uncovered dedicated neural pathways mediating the positive control of parenting, the regulation of infant-directed aggression and how it relates to adult-adult aggression is poorly understood. Here we show that urocortin-3 (Ucn3)-expressing neurons in the hypothalamic perifornical area (PeFAUcn3) are activated during infant-directed attacks in males and females, but not other behaviors. Functional manipulations of PeFAUcn3 neurons demonstrate the role of this population in the negative control of parenting in both sexes. PeFAUcn3 neurons receive input from areas associated with vomeronasal sensing, stress, and parenting, and send projections to hypothalamic and limbic areas. Optogenetic activation of PeFAUcn3 axon terminals in these regions triggers various aspects of infant-directed agonistic responses, such as neglect, repulsion and aggression. Thus, PeFAUcn3 neurons emerge as a dedicated circuit component controlling infant-directed neglect and aggression, providing a new framework to understand the positive and negative regulation of parenting in health and disease.
]]></description>
<dc:creator>Autry, A. E.</dc:creator>
<dc:creator>Wu, Z.</dc:creator>
<dc:creator>Kohl, J.</dc:creator>
<dc:creator>Bambah-Mukku, D.</dc:creator>
<dc:creator>Rubinstein, N. D.</dc:creator>
<dc:creator>Marin-Rodriguez, B.</dc:creator>
<dc:creator>Carta, I.</dc:creator>
<dc:creator>Sedwick, V.</dc:creator>
<dc:creator>Dulac, C.</dc:creator>
<dc:date>2019-07-09</dc:date>
<dc:identifier>doi:10.1101/697334</dc:identifier>
<dc:title><![CDATA[Perifornical Area Urocortin-3 Neurons Promote Infant-directed Neglect and Aggression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/697979v1?rss=1">
<title>
<![CDATA[
TRF1 prevents permissive DNA damage response, recombination and Break Induced Replication at telomeres 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/697979v1?rss=1"
</link>
<description><![CDATA[
Telomeres are a significant challenge to DNA replication and are prone to replication stress and telomere fragility. The shelterin component TRF1 facilitates telomere replication but the molecular mechanism remains uncertain. By interrogating the proteomic composition of telomeres, we show that telomeres lacking TRF1 undergo protein composition reorganisation associated with a DNA damage response and chromatin remodelers. Surprisingly, TRF1 suppresses the accumulation of promyelocytic leukemia (PML) protein, BRCA1 and the SMC5/6 complex at telomeres, which is associated with increased Homologous Recombination (HR) and TERRA transcription. We uncovered a previously unappreciated role for TRF1 in the suppression of telomere recombination, dependent on SMC5 and also POLD3 dependent Break Induced Replication at telomeres. We propose that TRF1 facilitates S-phase telomeric DNA synthesis to prevent illegitimate mitotic DNA recombination and chromatin rearrangement.
]]></description>
<dc:creator>Vannier, J.-B.</dc:creator>
<dc:creator>Porreca, R. M.</dc:creator>
<dc:creator>Law, P. P.</dc:creator>
<dc:creator>Herrera-Moyano, E.</dc:creator>
<dc:creator>Gonzalez Franco, R.</dc:creator>
<dc:creator>Montoya, A.</dc:creator>
<dc:creator>Faull, P.</dc:creator>
<dc:creator>Kramer, H.</dc:creator>
<dc:date>2019-07-10</dc:date>
<dc:identifier>doi:10.1101/697979</dc:identifier>
<dc:title><![CDATA[TRF1 prevents permissive DNA damage response, recombination and Break Induced Replication at telomeres]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/691972v1?rss=1">
<title>
<![CDATA[
Structure of D. melanogaster ARC1 reveals a repurposed molecule with characteristics of retroviral Gag 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/691972v1?rss=1"
</link>
<description><![CDATA[
The tetrapod neuronal protein ARC and its D. melanogaster homologue, dARC1, have important but differing roles in neuronal development. Both are thought to originate through exaptation of ancient Ty3/Gypsy retrotransposon Gag genes, with their novel function relying on an original capacity for self-assembly and encapsidation of nucleic acids. Here, we present the crystal structure of dARC1 CA and examine the relationship between dARC1, mammalian ARC and the CA protein of circulating retroviruses. We show that whilst the overall architecture is highly related to that of orthoretroviral and spumaretroviral CA, there are significant deviations in both N- and C-terminal domains, potentially affecting recruitment of partner proteins and particle assembly. The degree of sequence and structural divergence suggests that Ty3/Gypsy Gag has been exapted on two separate occasions and that, although mammalian ARC and dARC1 share functional similarity, the structures have undergone different adaptations after appropriation into the tetrapod and insect genomes.
]]></description>
<dc:creator>Cottee, M. A.</dc:creator>
<dc:creator>Letham, S. C.</dc:creator>
<dc:creator>Young, G. R.</dc:creator>
<dc:creator>Stoye, J. P.</dc:creator>
<dc:creator>Taylor, I. A.</dc:creator>
<dc:date>2019-07-10</dc:date>
<dc:identifier>doi:10.1101/691972</dc:identifier>
<dc:title><![CDATA[Structure of D. melanogaster ARC1 reveals a repurposed molecule with characteristics of retroviral Gag]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/710871v1?rss=1">
<title>
<![CDATA[
Lessons in genome engineering: opportunities, tools and pitfalls 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/710871v1?rss=1"
</link>
<description><![CDATA[
CRISPR/Cas technology allows the creation of double strand breaks and hence loss of function mutations at any location in the genome. This technology is now routine for many organisms and cell lines. Here we describe how CRISPR/Cas can be combined with other DNA manipulation techniques (e.g. homology-based repair, site-specific integration and Cre or FLP-mediated recombination) to create sophisticated tools to measure and manipulate gene activity. In one class of applications, a single site-specific insertion generates a transcriptional reporter, a loss-of function allele, and a tagged allele. In a second class of modifications, essential sequences are deleted and replaced with an integrase site, which serves as a platform for the creation of custom reporters, transcriptional drivers, conditional alleles and regulatory mutations. We describe how these tools and protocols can be implemented easily and efficiently. Importantly, we also highlight unanticipated failures, which serve as cautionary tales, and suggest mitigating measures. Our tools are designed for use in Drosophila but the lessons we draw are likely to be widely relevant.nnAUTHOR SUMMARYThe genome contains all the information that an organism needs to develop and function throughout its life. One of the goal of genetics is to decipher the role of all the genes (typically several thousands for an animal) present in the genome. One approach is to delete each gene and assay the consequences. Deletion of individual genes is now readily achieved with a technique called CRISPR/Cas9. However, simple genetic deletion provides limited information. Here we describe strains and DNA vectors that streamline the generation of more sophisticated genetic tools. We describe general means of creating alleles (genetic variants) that enable gene activity to be measured and experimentally modulated in space and time. Although the tools we describe are universally applicable, each gene requires special consideration. Based on our experience of successes and failures, we suggest measures to maximise the chances that engineered alleles serve their intended purpose. Although our methods are designed for use in Drosophila, they could be adapted to any organism that is amenable to CRISPR/Cas9 genome modification.
]]></description>
<dc:creator>Poernbacher, I.</dc:creator>
<dc:creator>Crossman, S.</dc:creator>
<dc:creator>Kurth, J.</dc:creator>
<dc:creator>Nojima, H.</dc:creator>
<dc:creator>Baena-Lopez, A.</dc:creator>
<dc:creator>Alexandre, C.</dc:creator>
<dc:creator>vincent, J.-P.</dc:creator>
<dc:date>2019-07-22</dc:date>
<dc:identifier>doi:10.1101/710871</dc:identifier>
<dc:title><![CDATA[Lessons in genome engineering: opportunities, tools and pitfalls]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/711259v1?rss=1">
<title>
<![CDATA[
A landmark-free morphometric pipeline: Application to characterise a Down syndrome model skull phenotype 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/711259v1?rss=1"
</link>
<description><![CDATA[
Characterising phenotypes often requires quantification of anatomical shapes. Quantitative shape comparison (morphometrics) traditionally uses anatomical landmarks and is therefore limited by the number of landmarks and operator accuracy when landmarks are located manually. Here we apply a landmark-free method to characterise the craniofacial skeletal phenotype of the Dp1Tyb mouse model of Down syndrome (DS), validating it against a landmark-based approach. We identify cranial dysmorphologies in Dp1Tyb mice, especially smaller size and brachycephaly (front-back shortening) homologous to the human phenotype. The landmark-free phenotyping was less labour-intensive and required less user training than the landmark-based method. It also enabled mapping of local differences as planar expansion or shrinkage. This higher resolution and local mapping pinpointed reductions in interior mid-snout structures and occipital bones in this DS model that were not as apparent using a traditional landmark-based method. This approach could make morphometrics widely-accessible beyond traditional niches in zoology and palaeontology, especially in characterising mutant phenotypes.
]]></description>
<dc:creator>Toussaint, N.</dc:creator>
<dc:creator>Redhead, Y.</dc:creator>
<dc:creator>Liu, W.</dc:creator>
<dc:creator>Fisher, E.</dc:creator>
<dc:creator>Hallgrimsson, B.</dc:creator>
<dc:creator>Tybulewicz, V.</dc:creator>
<dc:creator>Green, J. B. A.</dc:creator>
<dc:creator>Schnabel, J.</dc:creator>
<dc:date>2019-07-22</dc:date>
<dc:identifier>doi:10.1101/711259</dc:identifier>
<dc:title><![CDATA[A landmark-free morphometric pipeline: Application to characterise a Down syndrome model skull phenotype]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/714675v1?rss=1">
<title>
<![CDATA[
O-linked mucin-type glycosylation regulates the transcriptional programme downstream of EGFR in breast cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/714675v1?rss=1"
</link>
<description><![CDATA[
Aberrant mucin type O-linked glycosylation is a common occurrence in cancer. This type of O-linked glycosylation is not limited to mucins but can occur on many cell surface glycoproteins where only a small number of sites may be present. Upon EGF ligation, EGFR induces a signaling cascade but can also translocate to the nucleus where it can directly regulate gene transcription. Here we show that upon EGF binding, human breast cancer cells carrying different O-linked glycans respond by transcribing different gene expression signatures. This is not a result of changes in signal transduction but due to the differential nuclear translocation of EGFR in the two glyco-phenotypes. This is regulated by the formation of an EGFR/galectin-3/MUC1/{beta}-catenin complex at the cell surface that is present in cells carrying short core-1-based O-glycans characteristic of tumour cells but absent in core-2-carrying cells.
]]></description>
<dc:creator>Tajadura-Ortega, V.</dc:creator>
<dc:creator>Gambardella, G.</dc:creator>
<dc:creator>Skinner, A.</dc:creator>
<dc:creator>Ter-Borch Gram Schjoldager, K.</dc:creator>
<dc:creator>Beatson, R.</dc:creator>
<dc:creator>Graham, R.</dc:creator>
<dc:creator>Achkova, D.</dc:creator>
<dc:creator>Taylor-Papadimitriou, J.</dc:creator>
<dc:creator>Ciccarelli, F. D.</dc:creator>
<dc:creator>Burchell, J.</dc:creator>
<dc:date>2019-07-25</dc:date>
<dc:identifier>doi:10.1101/714675</dc:identifier>
<dc:title><![CDATA[O-linked mucin-type glycosylation regulates the transcriptional programme downstream of EGFR in breast cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/669861v1?rss=1">
<title>
<![CDATA[
Chemical precision glyco-mutagenesis by glycosyltransferase engineering in living cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/669861v1?rss=1"
</link>
<description><![CDATA[
Studying posttranslational modifications classically relies on experimental strategies that oversimplify the complex biosynthetic machineries of living cells. Protein glycosylation contributes to essential biological processes, but correlating glycan structure, underlying protein and disease-relevant biosynthetic regulation is currently elusive. Here, we engineer living cells to tag glycans with editable chemical functionalities while providing information on biosynthesis, physiological context and glycan fine structure. We introduce a non-natural substrate biosynthetic pathway and use engineered glycosyltransferases to incorporate chemically tagged sugars into the cell surface glycome of the living cell. We apply the strategy to a particularly redundant yet disease-relevant human glycosyltransferase family, the polypeptide N-acetylgalactosaminyl transferases. This approach bestows a gain-of-function modification on cells where the products of individual glycosyltransferases can be selectively characterized or manipulated at will.
]]></description>
<dc:creator>Schumann, B.</dc:creator>
<dc:creator>Malaker, S. A.</dc:creator>
<dc:creator>Wisnovsky, S. P.</dc:creator>
<dc:creator>Debets, M. F.</dc:creator>
<dc:creator>Agbay, A. J.</dc:creator>
<dc:creator>Fernandez, D.</dc:creator>
<dc:creator>Wagner, L. J. S.</dc:creator>
<dc:creator>Lin, L.</dc:creator>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>Fox, D. M.</dc:creator>
<dc:creator>Peh, J.</dc:creator>
<dc:creator>Gray, M. A.</dc:creator>
<dc:creator>Pedram, K.</dc:creator>
<dc:creator>Kohler, J. J.</dc:creator>
<dc:creator>Mrksich, M.</dc:creator>
<dc:creator>Bertozzi, C. R.</dc:creator>
<dc:date>2019-06-13</dc:date>
<dc:identifier>doi:10.1101/669861</dc:identifier>
<dc:title><![CDATA[Chemical precision glyco-mutagenesis by glycosyltransferase engineering in living cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/719062v1?rss=1">
<title>
<![CDATA[
Global profiling of myristoylation in Toxoplasma gondii reveals key roles for lipidation in CDPK1 and MIC7 function 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/719062v1?rss=1"
</link>
<description><![CDATA[
N-myristoylation is a ubiquitous class of protein lipidation across eukaryotes and N-myristoyl transferase has been proposed as an attractive drug target in several pathogens. Functionally the myristate often primes for subsequent palmitoylation and stable membrane attachment, however, growing evidence also suggests additional regulatory roles for myristoylation on proteins. Here we describe the first global chemoproteomic screening of protein myristoylation in Toxoplasma gondii. Through quantitative mass spectrometry coupled with validated chemoproteomic tools, we identify 65 myristoylated proteins. We report functionally important myristoylation on the key signalling protein CDPK1 and, surprisingly, myristoylation of the microneme protein 7 (MIC7), a predicted type-I-transmembrane protein. We demonstrate that myristoylation of MIC7 is not important for the trafficking to micronemes, but appears to play a role in host cell invasion. This dataset represents a large fraction of the parasites myristoylated proteome and a prerequisite to investigate this modification in Toxoplasma.
]]></description>
<dc:creator>Broncel, M.</dc:creator>
<dc:creator>Dominicus, C.</dc:creator>
<dc:creator>Hunt, A.</dc:creator>
<dc:creator>Wallbank, B. A.</dc:creator>
<dc:creator>Federico, S.</dc:creator>
<dc:creator>Young, J. C.</dc:creator>
<dc:creator>Treeck, M.</dc:creator>
<dc:date>2019-07-30</dc:date>
<dc:identifier>doi:10.1101/719062</dc:identifier>
<dc:title><![CDATA[Global profiling of myristoylation in Toxoplasma gondii reveals key roles for lipidation in CDPK1 and MIC7 function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/721043v1?rss=1">
<title>
<![CDATA[
Precision of Tissue Patterning is Controlled by Dynamical Properties of Gene Regulatory Networks 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/721043v1?rss=1"
</link>
<description><![CDATA[
During development, gene regulatory networks allocate cell fates by partitioning tissues into spatially organised domains of gene expression. How the sharp boundaries that delineate these gene expression patterns arise, despite the stochasticity associated with gene regulation, is poorly understood. We show, in the vertebrate neural tube, using perturbations of coding and regulatory regions, that the structure of the regulatory network contributes to boundary precision. This is achieved, not by reducing noise in individual genes, but by the configuration of the network modulating the ability of stochastic fluctuations to initiate gene expression changes. We use a computational screen to identify network properties that influence boundary precision, revealing two dynamical mechanisms by which small gene circuits attenuate the effect of noise in order to increase patterning precision. These results highlight design principles of gene regulatory networks that produce precise patterns of gene expression.
]]></description>
<dc:creator>Exelby, K.</dc:creator>
<dc:creator>Herrera-Delgado, E.</dc:creator>
<dc:creator>Garcia Perez, L.</dc:creator>
<dc:creator>Perez-Carrasco, R.</dc:creator>
<dc:creator>Sagner, A.</dc:creator>
<dc:creator>Metzis, V.</dc:creator>
<dc:creator>Sollich, P.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:date>2019-07-31</dc:date>
<dc:identifier>doi:10.1101/721043</dc:identifier>
<dc:title><![CDATA[Precision of Tissue Patterning is Controlled by Dynamical Properties of Gene Regulatory Networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/721548v1?rss=1">
<title>
<![CDATA[
jULIEs: extracellular probes for recordings and stimulation in the structurally and functionally intact mouse brain 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/721548v1?rss=1"
</link>
<description><![CDATA[
High signal-to-noise, scalable and minimally invasive recording and stimulation of the nervous system in intact animals is of fundamental importance to advance the understanding of brain function. Extracellular electrodes are among the most powerful tools capable of interfacing with large neuronal populations1-3. Neuronal tissue damage remains a major limiting factor in scaling electrode arrays, and has been found to correlate with electrode diameter across different electrode materials, such as microfabricated Michigan and Utah-style arrays4, MEMS and microsystems5, soft polymer or tungsten electrodes6 and Parylene C probes7. Small diameter ultramicroelectrodes (UMEs), while highly desirable, pose significant technical challenges such as reaching sufficient electrolyte-electrode coupling and limiting stray signal loss. To overcome these challenges, we have designed juxtacellular Ultra-Low Impedance Electrodes (jULIEs), a scalable technique for achieving high signal-to-noise electrical recordings as well as stimulation with UMEs. jULIEs are metal-glass composite UMEs thermally drawn to outer diameters (OD) of <25 {micro}m, with metal core diameters (ID) of as little as 1 {micro}m. We introduce a two-step electrochemical modification strategy that reduces UME coupling impedances by two orders of magnitude. Modifications enabled high signal-to-noise neural recordings in vivo through wires with micrometer scale core diameters. Histological and imaging experiments indicated that local vascular damage is minimal. Spikes reached amplitudes over 1 mV in vivo, indicating that recordings are possible in close proximity to intact neurons. Recording sites can be arranged in arbitrary patterns tailored to various neuroanatomical target structures and allowing parallel penetrations. jULIEs thus represent a versatile platform that allows for reliable recording and manipulation of neural activity in any areas of the functionally intact mammalian brain.
]]></description>
<dc:creator>Racz, R. R.</dc:creator>
<dc:creator>Kollo, M.</dc:creator>
<dc:creator>Racz, G.</dc:creator>
<dc:creator>Bulz, C.</dc:creator>
<dc:creator>Ackels, T.</dc:creator>
<dc:creator>Warner, T.</dc:creator>
<dc:creator>Wray, W. A.</dc:creator>
<dc:creator>Kiskin, N.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Ye, Z.</dc:creator>
<dc:creator>de Hoz, L.</dc:creator>
<dc:creator>Rancz, E. A.</dc:creator>
<dc:creator>Schaefer, A. T.</dc:creator>
<dc:date>2019-08-01</dc:date>
<dc:identifier>doi:10.1101/721548</dc:identifier>
<dc:title><![CDATA[jULIEs: extracellular probes for recordings and stimulation in the structurally and functionally intact mouse brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/730143v1?rss=1">
<title>
<![CDATA[
Improving protein function prediction with synthetic feature samples created by generative adversarial networks 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/730143v1?rss=1"
</link>
<description><![CDATA[
Protein function prediction is a challenging but important task in bioinformatics. Many prediction methods have been developed, but are still limited by the bottleneck on training sample quantity. Therefore, it is valuable to develop a data augmentation method that can generate high-quality synthetic samples to further improve the accuracy of prediction methods. In this work, we propose a novel generative adversarial networks-based method, namely FFPred-GAN, to accurately learn the high-dimensional distributions of protein sequence-based biophysical features and also generate high-quality synthetic protein feature samples. The experimental results suggest that the synthetic protein feature samples are successful in improving the prediction accuracy for all three domains of the Gene Ontology through augmentation of the original training protein feature samples.
]]></description>
<dc:creator>Wan, C.</dc:creator>
<dc:creator>Jones, D. T.</dc:creator>
<dc:date>2019-08-08</dc:date>
<dc:identifier>doi:10.1101/730143</dc:identifier>
<dc:title><![CDATA[Improving protein function prediction with synthetic feature samples created by generative adversarial networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/732263v1?rss=1">
<title>
<![CDATA[
Th22 cells are a major contributor to the mycobacterial CD4+ T cell response and are depleted during HIV infection 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/732263v1?rss=1"
</link>
<description><![CDATA[
HIV-1 infection substantially increases the risk of developing tuberculosis (TB). Some mechanisms, such as defects in the Th1 response to Mycobacterium tuberculosis (M.tb) in HIV-infected individuals have been widely reported. However, Th1-independent mechanisms also contribute to protection against TB. To identify a broader spectrum of defects in TB immunity during HIV infection, we examined IL-17 and IL-22 production in response to mycobacterial antigens in individuals with latent TB infection (LTBI) and HIV co-infection. Upon stimulating with mycobacterial antigens, we observed a distinct CD4+ T helper lineage producing IL-22 in the absence of IL-17 and IFN-{gamma}. Th22 cells were present at high frequencies in response to mycobacterial antigens in blood and contributed up to 50% to the CD4+ T cell response to mycobacteria, comparable in magnitude to the IFN-{gamma} Th1 response (median 0.91% and 0.55%, respectively). Phenotypic characterization of Th22 cells revealed that their memory differentiation was similar to M.tb-specific Th1 cells (i.e. predominantly early-differentiated CD45RO+CD27+ phenotype). Moreover, CCR6 and CXCR3 expression profiles of Th22 cells were similar to Th17 cells, while their CCR4 and CCR10 expression patterns displayed an intermediate phenotype between Th1 and Th17 cells. Strikingly, mycobacterial IL-22 responses were three-fold lower in HIV-infected individuals compared to uninfected individuals, and the magnitude of responses correlated inversely with HIV viral load. These data provide important insights into mycobacteria-specific T helper subsets and suggest a potential role for IL-22 in protection against TB during HIV infection. Further studies are needed to fully elucidate the role of IL-22 in protective TB immunity.
]]></description>
<dc:creator>Bunjun, R.</dc:creator>
<dc:creator>Omondi, F. M. A.</dc:creator>
<dc:creator>Makatsa, M. S.</dc:creator>
<dc:creator>Muller, T. L.</dc:creator>
<dc:creator>Prentice, C.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Riou, C.</dc:creator>
<dc:creator>Burgers, W. A.</dc:creator>
<dc:date>2019-08-13</dc:date>
<dc:identifier>doi:10.1101/732263</dc:identifier>
<dc:title><![CDATA[Th22 cells are a major contributor to the mycobacterial CD4+ T cell response and are depleted during HIV infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/735811v1?rss=1">
<title>
<![CDATA[
Lysine acetylation promotes PALB2 chromatin association to maintain genome stability 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/735811v1?rss=1"
</link>
<description><![CDATA[
The tumour suppressor PALB2 stimulates error-free repair of DNA breaks, whilst its steady-state chromatin association protects active genes from genotoxic stress. Here, we report that the lysine acetyltransferases 2A and 2B (KAT2A/B), commonly known to promote transcriptional activation, acetylate the PALB2 chromatin association motif (ChAM), providing a dynamic regulatory mechanism for PALB2. ChAM acetylation within a cluster of seven lysine residues (7K), detected in the chromatin-enriched fraction in undamaged cells, enhanced its association with nucleosomes while decreasing its non-specific binding to naked DNA. DNA damage triggered a rapid deacetylation of ChAM and a concomitant increase in PALB2 mobility. Significantly, a 7K-null mutation, which hindered ChAM binding to both nucleosomes and DNA, conferred deficiency in DNA repair and hypersensitivity to the anti-cancer drug olaparib. Thus, our study reveals a unique mechanism mediated by KAT2A/B-dependent acetylation of a non-histone protein, which fine-tunes the DNA damage response and hence promotes genome stability.
]]></description>
<dc:creator>Esashi, F.</dc:creator>
<dc:creator>Fournier, M.</dc:creator>
<dc:creator>Bleuyard, J.-Y.</dc:creator>
<dc:creator>Couturier, A. M.</dc:creator>
<dc:creator>Ellins, J.</dc:creator>
<dc:creator>Hester, S.</dc:creator>
<dc:creator>Smerdon, S. J.</dc:creator>
<dc:creator>Tora, L.</dc:creator>
<dc:date>2019-08-15</dc:date>
<dc:identifier>doi:10.1101/735811</dc:identifier>
<dc:title><![CDATA[Lysine acetylation promotes PALB2 chromatin association to maintain genome stability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/745141v1?rss=1">
<title>
<![CDATA[
A single cell transcriptome atlas of the developing zebrafish hindbrain 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/745141v1?rss=1"
</link>
<description><![CDATA[
Segmentation of the vertebrate hindbrain leads to the formation of rhombomeres, each with a distinct anteroposterior identity. Specialised boundary cells form at segment borders that act as a source or regulator of neuronal differentiation. In zebrafish, there is spatial patterning of neurogenesis in which non-neurogenic zones form at bounderies and segment centres, in part mediated by Fgf20 signaling. To further understand the control of neurogenesis, we have carried out single cell RNA sequencing of the zebrafish hindbrain at three different stages of patterning. Analyses of the data reveal known and novel markers of distinct hindbrain segments, of cell types along the dorsoventral axis, and of the transition of progenitors to neuronal differentiation. We find major shifts in the transcriptome of progenitors and of differentiating cells between the different stages analysed. Supervised clustering with markers of boundary cells and segment centres, together with RNA-seq analysis of Fgf-regulated genes, has revealed new candidate regulators of cell differentiation in the hindbrain. These data provide a valuable resource for functional investigations of the patterning of neurogenesis and the transition of progenitors to neuronal differentiation.
]]></description>
<dc:creator>Tambalo, M.</dc:creator>
<dc:creator>Mitter, R.</dc:creator>
<dc:creator>Wilkinson, D.</dc:creator>
<dc:date>2019-08-23</dc:date>
<dc:identifier>doi:10.1101/745141</dc:identifier>
<dc:title><![CDATA[A single cell transcriptome atlas of the developing zebrafish hindbrain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/755181v1?rss=1">
<title>
<![CDATA[
Elongations of epithelial colony in vitro: symmetry breaking through collective effects 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/755181v1?rss=1"
</link>
<description><![CDATA[
Epithelial tissues of the developing embryos elongate by different mechanisms, such as neighbor exchange, cell elongation, and oriented cell division. Since autonomous tissue self-organization is influenced by external cues such as morphogen gradients or neighboring tissues, it is difficult to distinguish intrinsic from directed tissue behavior. The mesoscopic processes leading to the different mechanisms remain elusive. Here, we study the spontaneous elongation behavior of spreading circular epithelial colonies in vitro. By quantifying deformation kinematics at multiple scales, we report that global elongation happens primarily due to cell elongations, and its direction correlates with the anisotropy of the average cell elongation. By imposing an external time-periodic stretch, the axis of this global symmetry breaking can be modified and elongation occurs primarily due to orientated neighbor exchange. These different behaviors are confirmed using a vertex model for collective cell behavior, providing a framework for understanding autonomous tissue elongation and its origins.
]]></description>
<dc:creator>Comelles, J.</dc:creator>
<dc:creator>Soumya, S. S.</dc:creator>
<dc:creator>Anvitha, S.</dc:creator>
<dc:creator>Salbreux, G.</dc:creator>
<dc:creator>Jülicher, F.</dc:creator>
<dc:creator>Inamdar, M. M.</dc:creator>
<dc:creator>Riveline, D.</dc:creator>
<dc:date>2019-09-04</dc:date>
<dc:identifier>doi:10.1101/755181</dc:identifier>
<dc:title><![CDATA[Elongations of epithelial colony in vitro: symmetry breaking through collective effects]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/754499v1?rss=1">
<title>
<![CDATA[
Apical length modulates dendritic excitability in L5 pyramidal neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/754499v1?rss=1"
</link>
<description><![CDATA[
Anatomical similarity across the neocortex has led to the common assumption that the circuitry is modular and performs stereotyped computations. Layer 5 pyramidal neurons (L5PNs) in particular are thought to be central to cortical computation because of their extensive arborisation and nonlinear dendritic operations. Here, we demonstrate that computations associated with dendritic Ca2+ plateaus in L5PNs vary substantially between the primary and secondary visual cortices. L5PNs in the secondary visual cortex show reduced dendritic excitability and smaller propensity for burst firing. This reduced excitability is correlated with shorter apical dendrites. Using numerical modelling, we uncover a universal principle underlying the influence of apical length on dendritic backpropagation and excitability, based on a Na+ channel-dependent broadening of backpropagating action potentials. In summary, we provide new insights into the modulation of dendritic excitability by apical dendrite length and show that the operational repertoire of L5 neurons is not universal throughout the brain.
]]></description>
<dc:creator>Galloni, A. R.</dc:creator>
<dc:creator>Laffere, A.</dc:creator>
<dc:creator>Rancz, E. A.</dc:creator>
<dc:date>2019-09-04</dc:date>
<dc:identifier>doi:10.1101/754499</dc:identifier>
<dc:title><![CDATA[Apical length modulates dendritic excitability in L5 pyramidal neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/755439v1?rss=1">
<title>
<![CDATA[
Protein ubiquitylation is essential for the schizont to merozoite transition in Plasmodium falciparum blood-stage development 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/755439v1?rss=1"
</link>
<description><![CDATA[
Ubiquitylation is a common post translational modification of eukaryotic proteins and in the human malaria parasite, Plasmodium falciparum (Pf) overall ubiquitylation increases in the transition from intracellular schizont to extracellular merozoite stages in the asexual blood stage cycle. Here, we identify specific ubiquitylation sites of protein substrates in three intracellular parasite stages and extracellular merozoites; a total of 1464 sites in 546 proteins were identified (data available via ProteomeXchange with identifier PXD014998). 469 ubiquitylated proteins were identified in merozoites compared with only 160 in the preceding intracellular schizont stage, indicating a large increase in protein ubiquitylation associated with merozoite maturation. Following merozoite invasion of erythrocytes, few ubiquitylated proteins were detected in the first intracellular ring stage but as parasites matured through trophozoite to schizont stages the extent of ubiquitylation increased. We identified commonly used ubiquitylation motifs and groups of ubiquitylated proteins in specific areas of cellular function, for example merozoite pellicle proteins involved in erythrocyte invasion, exported proteins, and histones. To investigate the importance of ubiquitylation we screened ubiquitin pathway inhibitors in a parasite growth assay and identified the ubiquitin activating enzyme (UBA1 or E1) inhibitor MLN7243 (TAK-243) to be particularly effective. This small molecule was shown to be a potent inhibitor of recombinant PfUBA1, and a structural homology model of MLN7243 bound to the parasite enzyme highlights avenues for the development of P. falciparum specific inhibitors. We created a genetically modified parasite with a rapamycin-inducible functional deletion of uba1; addition of either MLN7243 or rapamycin to the recombinant parasite line resulted in the same phenotype, with parasite development blocked at the late schizont stage. These results indicate that the intracellular target of MLN7243 is UBA1, and this activity is essential for the final differentiation of schizonts to merozoites. The ubiquitylation of many merozoite proteins and their disappearance in ring stages are consistent with the idea that ubiquitylation leads to their destruction via the proteasome once their function is complete following invasion, which would allow amino acid recycling in the period prior to the parasites elaboration of a new food vacuole.
]]></description>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Encheva, V.</dc:creator>
<dc:creator>Green, J. L.</dc:creator>
<dc:creator>Lasonder, E.</dc:creator>
<dc:creator>Prommaban, A.</dc:creator>
<dc:creator>Kunzelmann, S.</dc:creator>
<dc:creator>Christodoulou, E.</dc:creator>
<dc:creator>Grainger, M.</dc:creator>
<dc:creator>Truongvan, N.</dc:creator>
<dc:creator>Bothe, S.</dc:creator>
<dc:creator>Sharma, V.</dc:creator>
<dc:creator>Song, W.</dc:creator>
<dc:creator>Pinzuti, I.</dc:creator>
<dc:creator>Uthaipibull, C.</dc:creator>
<dc:creator>Srichairatanakool, S.</dc:creator>
<dc:creator>Barault, V.</dc:creator>
<dc:creator>Langsley, G.</dc:creator>
<dc:creator>Schindelin, H.</dc:creator>
<dc:creator>Stieglitz, B.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:date>2019-09-04</dc:date>
<dc:identifier>doi:10.1101/755439</dc:identifier>
<dc:title><![CDATA[Protein ubiquitylation is essential for the schizont to merozoite transition in Plasmodium falciparum blood-stage development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/758318v1?rss=1">
<title>
<![CDATA[
The antibiotic bedaquiline activates host macrophage innate immune resistance to bacterial infection 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/758318v1?rss=1"
</link>
<description><![CDATA[
Antibiotics are widely used in the treatment of bacterial infections. Although known for their microbicidal activity, antibiotics may also interfere with the hosts immune system. Here we analyzed the effects of bedaquiline (BDQ), an inhibitor of the mycobacterial ATP synthase, on human macrophages. Genome-wide gene expression analysis revealed that BDQ reprogramed macrophages into potent bactericidal phagocytes. We found that 1,495 genes were differentially expressed in M. tuberculosis-infected macrophages incubated with the drug, with an over-representation of genes involved in metabolism, lysosome biogenesis and activation. BDQ treatment triggered a variety of antimicrobial defense mechanisms, including nitric oxide production, phagosome-lysosome fusion, and autophagy. These effects were associated with activation of transcription factor EB (TFEB), involved in the transcription of lysosomal genes, resulting in enhanced intracellular killing of different bacterial species that were naturally insensitive to BDQ. Thus, BDQ could be used as a host-directed therapy against a wide range of bacterial infections.
]]></description>
<dc:creator>Giraud-Gatineau, A.</dc:creator>
<dc:creator>Coya, J. M.</dc:creator>
<dc:creator>Maure, A.</dc:creator>
<dc:creator>Biton, A.</dc:creator>
<dc:creator>Thomson, M.</dc:creator>
<dc:creator>Bernard, E. M.</dc:creator>
<dc:creator>Gutierrez, M. G.</dc:creator>
<dc:creator>Larrouy-Maumus, G.</dc:creator>
<dc:creator>Brosch, R.</dc:creator>
<dc:creator>Gicquel, B.</dc:creator>
<dc:creator>Tailleux, L.</dc:creator>
<dc:date>2019-09-06</dc:date>
<dc:identifier>doi:10.1101/758318</dc:identifier>
<dc:title><![CDATA[The antibiotic bedaquiline activates host macrophage innate immune resistance to bacterial infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/762146v1?rss=1">
<title>
<![CDATA[
Delineating the rules for structural adaptation of membrane-associated proteins to evolutionary changes in membrane lipidome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/762146v1?rss=1"
</link>
<description><![CDATA[
Membrane function is fundamental to life. Each species explores membrane lipid diversity within a genetically predefined range of possibilities. How membrane lipid composition in turn defines the functional space available for evolution of membrane-centered processes remains largely unknown. We address this fundamental question using related fission yeasts Schizosaccharomyces pombe and Schizosaccharomyces japonicus. We show that unlike S. pombe that generates membranes where both glycerophospholipid acyl tails are predominantly 16-18 carbons long, S. japonicus synthesizes unusual  asymmetrical glycerophospholipids where the tails differ in length by 6-8 carbons. This results in stiffer bilayers with distinct lipid packing properties. Retroengineered S. pombe synthesizing the S. japonicus-type phospholipids exhibits unfolded protein response and downregulates secretion. Importantly, our protein sequence comparisons and domain swap experiments indicate that transmembrane helices co-evolve with membranes, suggesting that, on the evolutionary scale, changes in membrane lipid composition may necessitate extensive adaptation of the membrane-associated proteome.
]]></description>
<dc:creator>Makarova, M.</dc:creator>
<dc:creator>Peter, M.</dc:creator>
<dc:creator>Balogh, G.</dc:creator>
<dc:creator>Glatz, A.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Mora, N. L.</dc:creator>
<dc:creator>Booth, P.</dc:creator>
<dc:creator>Makeyev, E.</dc:creator>
<dc:creator>Vigh, L.</dc:creator>
<dc:creator>Oliferenko, S.</dc:creator>
<dc:date>2019-09-09</dc:date>
<dc:identifier>doi:10.1101/762146</dc:identifier>
<dc:title><![CDATA[Delineating the rules for structural adaptation of membrane-associated proteins to evolutionary changes in membrane lipidome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/764670v1?rss=1">
<title>
<![CDATA[
Homeostatic maintenance and age-related functional decline in the Drosophila ear 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/764670v1?rss=1"
</link>
<description><![CDATA[
The widespread loss of hearing is one of the major threats to future wellbeing in ageing human societies. Amongst its various forms, age-related hearing loss (ARHL) carries the vast bulk of the global disease burden. The causes for the terminal decline of auditory function, however, are as unknown as the mechanisms that maintain sensitive hearing before its breakdown. We here present an in-depth analysis of maintenance and ageing in the auditory system of the fruit fly Drosophila melanogaster. We show that Drosophila, just like humans, display ARHL and that their auditory life span is homeostatically supported by a set of evolutionarily conserved transcription factors. The transcription factors Onecut (closest human orthologues: ONECUT2, ONECUT3), Optix (SIX3, SIX6), Worniu (SNAI2) and Amos (ATOH1, ATOH7, NEUROD1) emerged as key regulators acting upstream of core sensory genes, including components of the flys molecular machinery for auditory transduction and amplification.
]]></description>
<dc:creator>Keder, A.</dc:creator>
<dc:creator>Tardieu, C.</dc:creator>
<dc:creator>Malong, L.</dc:creator>
<dc:creator>Filia, A.</dc:creator>
<dc:creator>Kashkenbayeva, A.</dc:creator>
<dc:creator>Gale, J. E.</dc:creator>
<dc:creator>Lovett, M.</dc:creator>
<dc:creator>Jarman, A. P.</dc:creator>
<dc:creator>Albert, J. T.</dc:creator>
<dc:date>2019-09-10</dc:date>
<dc:identifier>doi:10.1101/764670</dc:identifier>
<dc:title><![CDATA[Homeostatic maintenance and age-related functional decline in the Drosophila ear]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/767830v1?rss=1">
<title>
<![CDATA[
Real-time dynamics of Plasmodium NDC80 as a marker for the kinetochore during atypical mitosis and meiosis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/767830v1?rss=1"
</link>
<description><![CDATA[
Eukaryotic cell proliferation requires chromosome replication and precise segregation to ensure daughter cells have identical genomic copies. The genus Plasmodium, the causative agent of malaria, displays remarkable aspects of nuclear division throughout its lifecycle to meet some peculiar and unique challenges of DNA replication and chromosome segregation. The parasite undergoes atypical endomitosis and endoreduplication with an intact nuclear membrane and intranuclear mitotic spindle. To understand these diverse modes of Plasmodium cell division, we have studied the behaviour and composition of the outer kinetochore NDC80 complex, a key part of the mitotic apparatus that attaches the centromere of chromosomes to microtubules of the mitotic spindle. Using NDC80-GFP live-cell imaging in Plasmodium berghei we observe dynamic spatiotemporal changes during proliferation, including highly unusual kinetochore arrangements during sexual stages. We identify a very divergent candidate for the SPC24 subunit of the NDC80 complex, previously thought to be missing in Plasmodium, which completes a canonical, albeit unusual, NDC80 complex structure. Altogether, our studies reveal the kinetochore as an ideal tool to investigate the non-canonical modes of chromosome segregation and cell division in Plasmodium.

Summary StatementThe dynamic localization of kinetochore marker NDC80 protein complex during proliferative stages of the malaria parasite life cycle reveals unique modes of chromosome segregation.
]]></description>
<dc:creator>Pandey, R.</dc:creator>
<dc:creator>Zeeshan, M.</dc:creator>
<dc:creator>Ferguson, D. J. P.</dc:creator>
<dc:creator>Markus, R.</dc:creator>
<dc:creator>Brady, D.</dc:creator>
<dc:creator>Daniel, E.</dc:creator>
<dc:creator>Stanway, R. R.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:creator>Guttery, D. S.</dc:creator>
<dc:creator>Tewari, R.</dc:creator>
<dc:date>2019-09-13</dc:date>
<dc:identifier>doi:10.1101/767830</dc:identifier>
<dc:title><![CDATA[Real-time dynamics of Plasmodium NDC80 as a marker for the kinetochore during atypical mitosis and meiosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/769356v1?rss=1">
<title>
<![CDATA[
Copy number motifs expose genome instability type and predict driver events and disease outcome in breast cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/769356v1?rss=1"
</link>
<description><![CDATA[
Tumor evolution is dependent on and constrained by the genotypes emerging from genome instability. We hypothesized that non-site-specific copy number motifs would correlate with underlying replication defects and also with tumor and patient fate. Six feature detectors were defined to characterize and score the local spatial behaviour of a copy number profile. By accumulating scores across genomic regions, a low-dimensional representation of the tumor genome was obtained. The proposed Copy Aberration Regional Mapping Analysis (CARMA) algorithm was applied to 2384 breast tumors from three breast cancer cohorts, revealing distinct copy number motifs in established molecular subtypes. A prognostic index combining the features predicted breast cancer specific survival better than both the genomic instability index (GII) and all commonly used clinical stratifications. CARMA offers effective comparison of tumor subgroups and extracts biologically and clinically relevant features from allele-specific copy number profiles.
]]></description>
<dc:creator>Pladsen, A. V.</dc:creator>
<dc:creator>Nilsen, G.</dc:creator>
<dc:creator>Rueda, O. M.</dc:creator>
<dc:creator>Aure, M. R.</dc:creator>
<dc:creator>Borgan, O.</dc:creator>
<dc:creator>Liestol, K.</dc:creator>
<dc:creator>Vitelli, V.</dc:creator>
<dc:creator>Frigessi, A.</dc:creator>
<dc:creator>Langerod, A.</dc:creator>
<dc:creator>OSBREAC,</dc:creator>
<dc:creator>Mathelier, A.</dc:creator>
<dc:creator>Engebraten, O.</dc:creator>
<dc:creator>Wedge, D.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>Caldas, C.</dc:creator>
<dc:creator>Borresen-Dale, A.-L.</dc:creator>
<dc:creator>Russnes, H. G.</dc:creator>
<dc:creator>Lingjaerde, O. C.</dc:creator>
<dc:date>2019-09-14</dc:date>
<dc:identifier>doi:10.1101/769356</dc:identifier>
<dc:title><![CDATA[Copy number motifs expose genome instability type and predict driver events and disease outcome in breast cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/770768v1?rss=1">
<title>
<![CDATA[
A natural variant of the sole pyruvate kinase of fission yeast lowers glycolytic flux triggering increased respiration and oxidative-stress resistance but decreased growth 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/770768v1?rss=1"
</link>
<description><![CDATA[
Cells balance glycolysis with respiration to support their energetic and biosynthetic needs in different environmental or physiological contexts. With abundant glucose, many cells prefer to grow by aerobic glycolysis, or fermentation in yeast. Using 161 natural isolates of fission yeast, we investigated the genetic basis and phenotypic effects of the fermentation-respiration balance. The laboratory and a few other strains were more dependent on respiration. This trait was associated with a missense variant in a highly conserved region of Pyk1. Pyk1 is the single pyruvate kinase in fission yeast, while most organisms possess isoforms with different activity. This variant reduced Pyk1 activity and glycolytic flux. Replacing the  low-activity pyk1 allele in the laboratory strain with the common  high-activity allele was sufficient to increase fermentation and decrease respiration. This metabolic reprogramming triggered systems-level adaptations in the transcriptome and proteome, and in cellular phenotypes, including increased growth and chronological lifespan, but decreased resistance to oxidative stress. Thus, low Pyk1 activity provided no growth advantage but stress tolerance, despite increased respiration. The genetic tuning of glycolytic flux by a single-nucleotide change might reflect an adaptive trade-off in a species lacking pyruvate-kinase isoforms.
]]></description>
<dc:creator>Kamrad, S.</dc:creator>
<dc:creator>Grossbach, J.</dc:creator>
<dc:creator>Rodriguez-Lopez, M.</dc:creator>
<dc:creator>Townsend, S.</dc:creator>
<dc:creator>Mulleder, M.</dc:creator>
<dc:creator>Cappelletti, V.</dc:creator>
<dc:creator>Stojanovski, G.</dc:creator>
<dc:creator>Picotti, P.</dc:creator>
<dc:creator>Beyer, A.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:creator>Bahler, J.</dc:creator>
<dc:date>2019-09-16</dc:date>
<dc:identifier>doi:10.1101/770768</dc:identifier>
<dc:title><![CDATA[A natural variant of the sole pyruvate kinase of fission yeast lowers glycolytic flux triggering increased respiration and oxidative-stress resistance but decreased growth]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/772947v1?rss=1">
<title>
<![CDATA[
Fluctuation-based Super-Resolution Traction Force Microscopy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/772947v1?rss=1"
</link>
<description><![CDATA[
Cellular mechanics play a crucial role in tissue morphogenesis and homeostasis and are often misregulated in disease. Traction force microscopy (TFM) is one of the key methods that has enabled researchers to study fundamental aspects of mechanobiology; however, the power of TFM is limited by poor resolution and low throughput. Here, we propose a simplified protocol and imaging strategy, relying on super-resolution microscopy enabled by fluorophore fluctuation analysis, to enhance the output of TFM, by increasing both bead density as well as the accuracy of bead tracking in TFM gels. Our analysis pipeline can be used on either camera-based confocal or widefield microscopes and is fully compatible with available TFM analysis software. In addition, we demonstrate that our workflow can be used to gain biologically relevant information and is suitable for long-term live measurement of traction forces even in light-sensitive cells. Finally, we propose that our strategy could be used to considerably simplify the implementation of TFM screens. Our streamlined protocol can be performed with minimal hardware and software investment, and has the potential to standardize high-resolution TFM.
]]></description>
<dc:creator>Stubb, A.</dc:creator>
<dc:creator>Laine, R. F.</dc:creator>
<dc:creator>Guzman, C.</dc:creator>
<dc:creator>Henriques, R.</dc:creator>
<dc:creator>Jacquemet, G.</dc:creator>
<dc:creator>Ivaska, J.</dc:creator>
<dc:date>2019-09-18</dc:date>
<dc:identifier>doi:10.1101/772947</dc:identifier>
<dc:title><![CDATA[Fluctuation-based Super-Resolution Traction Force Microscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/774273v1?rss=1">
<title>
<![CDATA[
Proteasome-mediated protein degradation resets the cell division cycle and triggers ESCRT-III-mediated cytokinesis in an archaeon 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/774273v1?rss=1"
</link>
<description><![CDATA[
The archaeon Sulfolobus acidocaldarius is a relative of eukaryotes known to progress orderly through its cell division cycle despite lacking obvious CDK/cyclin homologues. Here, in exploring the mechanisms underpinning archaeal cell division cycle control, we show that the proteasome of S. acidocaldarius, like its eukaryotic counterpart, regulates the transition from the end of one cell division cycle to the beginning of the next. Further, we identify the archaeal ESCRT-III homologue CdvB as a key target of the proteasome, and show that state-dependent degradation of CdvB triggers archaeal cell division by allowing constriction of a CdvB1:CdvB2 ESCRT-III division ring. These findings suggest an ancient role for proteasome-mediated degradation in resetting the cell division cycle in both archaea and eukaryotes.
]]></description>
<dc:creator>Tarrason Risa, G.</dc:creator>
<dc:creator>Hurtig, F.</dc:creator>
<dc:creator>Bray, S.</dc:creator>
<dc:creator>Hafner, A.</dc:creator>
<dc:creator>Harker-Kirschneck, L.</dc:creator>
<dc:creator>Faull, P.</dc:creator>
<dc:creator>Davis, C.</dc:creator>
<dc:creator>Papatziamou, D.</dc:creator>
<dc:creator>Mutavchiev, D. R.</dc:creator>
<dc:creator>Fan, C.</dc:creator>
<dc:creator>Meneguello, L.</dc:creator>
<dc:creator>Arashiro Pulschen, A.</dc:creator>
<dc:creator>Dey, G.</dc:creator>
<dc:creator>Culley, S.</dc:creator>
<dc:creator>Kilkenny, M. L.</dc:creator>
<dc:creator>Pellegrini, L.</dc:creator>
<dc:creator>de Bruin, R. A. M.</dc:creator>
<dc:creator>Henriques, R.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Saric, A.</dc:creator>
<dc:creator>Lindas, A.-C.</dc:creator>
<dc:creator>Robinson, N.</dc:creator>
<dc:creator>Baum, B.</dc:creator>
<dc:date>2019-09-18</dc:date>
<dc:identifier>doi:10.1101/774273</dc:identifier>
<dc:title><![CDATA[Proteasome-mediated protein degradation resets the cell division cycle and triggers ESCRT-III-mediated cytokinesis in an archaeon]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/777326v1?rss=1">
<title>
<![CDATA[
Lamellipodin tunes cell migration by stabilizing protrusions and promoting adhesion formation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/777326v1?rss=1"
</link>
<description><![CDATA[
Efficient migration on adhesive surfaces involves the protrusion of lamellipodial actin networks and their subsequent stabilization by nascent adhesions. The actin binding protein lamellipodin (Lpd) is thought to play a critical role in lamellipodium protrusion, by delivering Ena/VASP proteins onto the growing plus ends of actin filaments and by interacting with the WAVE regulatory complex (WRC), an activator of the Arp2/3 complex, at the leading edge. Using B16-F1 melanoma cell lines, we demonstrate that genetic ablation of Lpd compromises protrusion efficiency and coincident cell migration without altering essential parameters of lamellipodia, including their maximal rate of forward advancement and actin polymerization. We also confirmed lamellipodia and migration phenotypes with CRISPR/Cas9-mediated Lpd knockout Rat2 fibroblasts, excluding cell type-specific effects. Moreover, computer-aided analysis of cell edge morphodynamics on B16-F1 cell lamellipodia revealed that loss of Lpd correlates with reduced temporal protrusion maintenance as a prerequisite of nascent adhesion formation. We conclude that Lpd optimizes protrusion and nascent adhesion formation by counteracting frequent, chaotic retraction and membrane ruffling.

Summary statementWe describe how genetic ablation of the prominent actin- and VASP-binding protein lamellipodin combined with software-aided protrusion analysis uncovers mechanistic insights into its cellular function during cell migration.
]]></description>
<dc:creator>Dimchev, G.</dc:creator>
<dc:creator>Amiri, B.</dc:creator>
<dc:creator>Humphries, A. C.</dc:creator>
<dc:creator>Schaks, M.</dc:creator>
<dc:creator>Dimchev, V.</dc:creator>
<dc:creator>Stradal, T. E. B.</dc:creator>
<dc:creator>Faix, J.</dc:creator>
<dc:creator>Krause, M.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:creator>Falcke, M.</dc:creator>
<dc:creator>Rottner, K.</dc:creator>
<dc:date>2019-09-20</dc:date>
<dc:identifier>doi:10.1101/777326</dc:identifier>
<dc:title><![CDATA[Lamellipodin tunes cell migration by stabilizing protrusions and promoting adhesion formation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/779108v1?rss=1">
<title>
<![CDATA[
The speed of GTP hydrolysis determines GTP cap size and controls microtubule stability 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/779108v1?rss=1"
</link>
<description><![CDATA[
Microtubules are bistable cytoskeletal polymers whose function depends on their property to switch between states of growth and shrinkage 1. Growing microtubules are thought to be stabilized by a GTP cap at their ends 2-5. The nature of this cap, however, is still poorly understood. How GTP hydrolysis determines the properties of the GTP cap and hence microtubule stability is unclear. End Binding proteins (EBs) recruit a diverse range of regulators of microtubule function to growing microtubule ends 6. Whether these regulatory platforms at growing microtubule ends are identical to the GTP cap is not known. Using mutated human tubulin with blocked GTP hydrolysis, we demonstrate in microscopy-based in vitro reconstitutions that EB proteins bind with high affinity to the GTP conformation of microtubules. Slowing-down GTP hydrolysis leads to extended GTP caps and consequently hyper-stable microtubules. Single molecule experiments reveal that the microtubule conformation gradually changes in the cap as GTP is hydrolyzed. These results demonstrate the critical importance of the kinetics of GTP hydrolysis for microtubule stability; and establish that the GTP cap coincides with the EB-binding regulatory hub that modulates microtubule cytoskeleton function in cells.
]]></description>
<dc:creator>Roostalu, J.</dc:creator>
<dc:creator>Thomas, C.</dc:creator>
<dc:creator>Cade, N. I.</dc:creator>
<dc:creator>Kunzelmann, S.</dc:creator>
<dc:creator>Taylor, I. A.</dc:creator>
<dc:creator>Surrey, T.</dc:creator>
<dc:date>2019-09-23</dc:date>
<dc:identifier>doi:10.1101/779108</dc:identifier>
<dc:title><![CDATA[The speed of GTP hydrolysis determines GTP cap size and controls microtubule stability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/779769v1?rss=1">
<title>
<![CDATA[
Closed mitosis requires local disassembly of the nuclear envelope 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/779769v1?rss=1"
</link>
<description><![CDATA[
At the end of mitosis, eukaryotic cells must segregate both copies of their replicated genome into two new nuclear compartments (1). They do this either by first dismantling and later reassembling the nuclear envelope in a so called "open mitosis", or by reshaping an intact nucleus and then dividing into two in a "closed mitosis" (2, 3). However, while mitosis has been studied in a wide variety of eukaryotes for over a century (4), it is not known how the double membrane of the nuclear envelope is split into two at the end of a closed mitosis without compromising the impermeability of the nuclear compartment (5). In studying this problem in the fission yeast Schizosaccharomyces pombe, a classical model for closed mitosis (5), we use genetics, live cell imaging and electron tomography to show that nuclear fission is achieved via local disassembly of the nuclear envelope (NE) within the narrow bridge that links segregating daughter nuclei. In doing so, we identify a novel inner NE-localised protein Les1 that restricts the process of local NE breakdown (local NEB) to the bridge midzone and prevents the leakage of material from daughter nuclei. The mechanics of local NEB in a closed mitosis closely mirror those of NEB in open mitosis (3), revealing an unexpectedly deep conservation of nuclear remodelling mechanisms across diverse eukaryotes.
]]></description>
<dc:creator>Dey, G.</dc:creator>
<dc:creator>Culley, S.</dc:creator>
<dc:creator>Curran, S. A.</dc:creator>
<dc:creator>Henriques, R.</dc:creator>
<dc:creator>Kukulski, W.</dc:creator>
<dc:creator>Baum, B.</dc:creator>
<dc:date>2019-09-23</dc:date>
<dc:identifier>doi:10.1101/779769</dc:identifier>
<dc:title><![CDATA[Closed mitosis requires local disassembly of the nuclear envelope]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/789578v1?rss=1">
<title>
<![CDATA[
Single-molecule imaging reveals control of parental histone recycling by free histones during DNA replication 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/789578v1?rss=1"
</link>
<description><![CDATA[
Faithful replication of chromatin domains during cell division is fundamental to eukaryotic development. During replication, nucleosomes are disrupted ahead of the replication fork, followed by their rapid reassembly on daughter strands from the pool of recycled parental and newly synthesized histones. Here, we use single-molecule imaging and replication assays in Xenopus laevis egg extracts to determine the outcome of replication fork encounters with nucleosomes. Contrary to current models, the majority of parental histones are evicted from the DNA, with histone recycling, nucleosome sliding and replication fork stalling also occurring but at lower frequencies. The anticipated local histone transfer only becomes dominant upon depletion of free histones from extracts. Our studies provide the first direct evidence that parental histones remain in close proximity to their original locus during recycling and reveal that provision of excess histones results in impaired histone recycling, which has the potential to affect epigenetic memory.
]]></description>
<dc:creator>Gruszka, D. T.</dc:creator>
<dc:creator>Xie, S.</dc:creator>
<dc:creator>Kimura, H.</dc:creator>
<dc:creator>Yardimci, H.</dc:creator>
<dc:date>2019-10-01</dc:date>
<dc:identifier>doi:10.1101/789578</dc:identifier>
<dc:title><![CDATA[Single-molecule imaging reveals control of parental histone recycling by free histones during DNA replication]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/792481v1?rss=1">
<title>
<![CDATA[
B cells extract antigens using Arp2/3-generated actin foci interspersed with linear filaments 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/792481v1?rss=1"
</link>
<description><![CDATA[
Antibody production depends on B cell internalization and presentation of antigens to helper T cells. To acquire antigens displayed by antigen-presenting cells, B cells form immune synapses and extract antigens by the mechanical activity of the acto-myosin cytoskeleton. While cytoskeleton organization driving the initial formation of the B cell synapse has been studied, how the cytoskeleton supports antigen extraction remains poorly understood. Here we show that after initial cell spreading, F-actin in B cell synapses forms a highly dynamic pattern composed of actin foci interspersed with linear filaments and myosin IIa. The foci are generated by Arp2/3-mediated branched-actin polymerization and stochastically associate with antigen clusters to mediate internalization. However, antigen extraction also requires the activity of formins, which reside near the foci and produce the interspersed filaments. Thus, a cooperation of branched-actin foci supported by linear filaments underlies B cell mechanics during antigen extraction.
]]></description>
<dc:creator>Roper, S. I.</dc:creator>
<dc:creator>Wasim, L.</dc:creator>
<dc:creator>Malinova, D.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:creator>Cox, S.</dc:creator>
<dc:creator>Tolar, P.</dc:creator>
<dc:date>2019-10-04</dc:date>
<dc:identifier>doi:10.1101/792481</dc:identifier>
<dc:title><![CDATA[B cells extract antigens using Arp2/3-generated actin foci interspersed with linear filaments]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/792804v1?rss=1">
<title>
<![CDATA[
Differential spatiotemporal targeting of Toxoplasma and Salmonella by GBP1 assembles caspase signalling platforms 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/792804v1?rss=1"
</link>
<description><![CDATA[
Guanylate binding proteins (GBPs), a family of interferon (IFN)-inducible GTPases, can promote cell-intrinsic defense by removal of intracellular microbial replicative niches through host cell death. GBPs target pathogen-containing vacuoles or the pathogen itself, and assist in membrane-disruption and release of microbial molecules that trigger cell death by activating the inflammasomes. We previously showed that GBP1 mediates atypical apoptosis or pyroptosis of human macrophages infected with Toxoplasma gondii (Tg) or Salmonella enterica Typhimurium (STm), respectively. In mice, the p47 Immunity-related GTPases (IRGs) control the recruitment of GBPs to microbe-containing vacuoles and subsequent cell death. However, humans are devoid of functional IRGs, and the pathogen-proximal immune detection mechanisms by GBP1 are poorly understood. Here, we describe two novel single-cell assays which show that GBP1 promotes the lysis of Tg-containing vacuoles and Tg plasma membrane, resulting in the cytosolic detection of Tg-DNA. In contrast, we show GBP1 only targets cytosolic STm and does not contribute to bacterial escape into the cytosol of human macrophages. GBP1 interacts with caspase-4 and recruits it directly to the bacterial surface, where caspase-4 can be activated by LPS. During STm infection, caspase-1 cleaves and inactivates GBP1 at Asp192, a site conserved in related mammalian GBP1 proteins but not in murine Gbps. STm-infected human macrophages expressing a cleavage-deficient GBP1 mutant exhibit higher pyroptosis due to the absence of caspase-1-mediated feedback inhibition of the GBP1-caspase-4 pathway. Our comparative studies elucidate microbe-specific spatiotemporal roles of GBP1 in detecting infection and the assembly and regulation of divergent caspase signaling platforms.
]]></description>
<dc:creator>Fisch, D.</dc:creator>
<dc:creator>Clough, B.</dc:creator>
<dc:creator>Domart, M.-C.</dc:creator>
<dc:creator>Bando, H.</dc:creator>
<dc:creator>Masonou, T.</dc:creator>
<dc:creator>Collinson, L. M.</dc:creator>
<dc:creator>Yamamoto, M.</dc:creator>
<dc:creator>Shenoy, A.</dc:creator>
<dc:creator>Frickel, E.-M.</dc:creator>
<dc:date>2019-10-05</dc:date>
<dc:identifier>doi:10.1101/792804</dc:identifier>
<dc:title><![CDATA[Differential spatiotemporal targeting of Toxoplasma and Salmonella by GBP1 assembles caspase signalling platforms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/796003v1?rss=1">
<title>
<![CDATA[
Mechanism of RPA-Facilitated Processive DNA Unwinding by the Eukaryotic CMG Helicase 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/796003v1?rss=1"
</link>
<description><![CDATA[
The DNA double helix is unwound by the Cdc45/Mcm2-7/GINS (CMG) complex at the eukaryotic replication fork. While isolated CMG unwinds duplex DNA very slowly, its fork unwinding rate is stimulated by an order of magnitude by single-stranded DNA binding protein, RPA. However, the molecular mechanism by which RPA enhances CMG helicase activity remained elusive. Here, we demonstrate that engagement of CMG with parental double-stranded DNA (dsDNA) at the replication fork impairs its helicase activity, explaining the slow DNA unwinding by isolated CMG. Using single-molecule and ensemble biochemistry, we show that binding of RPA to the excluded DNA strand prevents duplex engagement by the helicase and speeds up CMG-mediated DNA unwinding. When stalled due to dsDNA interaction, DNA rezipping-induced helicase backtracking re-establishes productive helicase-fork engagement underscoring the significance of plasticity in helicase action. Together, our results elucidate the dynamics of CMG at the replication fork and reveal how other replisome components can mediate proper DNA engagement by the replicative helicase to achieve efficient fork progression.
]]></description>
<dc:creator>Kose, H. B.</dc:creator>
<dc:creator>Xie, S.</dc:creator>
<dc:creator>Cameron, G.</dc:creator>
<dc:creator>Strycharska, M. S.</dc:creator>
<dc:creator>Yardimci, H.</dc:creator>
<dc:date>2019-10-07</dc:date>
<dc:identifier>doi:10.1101/796003</dc:identifier>
<dc:title><![CDATA[Mechanism of RPA-Facilitated Processive DNA Unwinding by the Eukaryotic CMG Helicase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/796524v1?rss=1">
<title>
<![CDATA[
Cryo-EM Structures of the XPF-ERCC1 Endonuclease Reveal an Auto-Inhibited Conformation and the Basis for Activation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/796524v1?rss=1"
</link>
<description><![CDATA[
The structure-specific endonuclease XPF-ERCC1 participates in multiple DNA damage repair pathways including nucleotide excision repair (NER) and inter-strand crosslink repair (ICLR). How XPF-ERCC1 is catalytically activated by DNA junction substrates is not currently understood. We report cryo-electron microscopy structures of both DNA-free and DNA-bound human XPF-ERCC1. DNA-free XPF-ERCC1 adopts an auto-inhibited conformation in which the XPF helical domain masks ERCC1 DNA-binding elements and restricts access to the XPF catalytic site. Binding of a model DNA junction separates the XPF helical and ERCC1 (HhH)2 domains, promoting activation. Using these structural data, we propose a model for a 5-NER incision complex involving XPF-ERCC1-XPA and a DNA junction substrate. Structure-function data suggest xeroderma pigmentosum patient mutations often compromise the structural integrity of XPF-ERCC1. Fanconi anaemia patient mutations often display substantial in-vitro activity but are resistant to activation by ICLR recruitment factor SLX4. Our data provide insights into XPF-ERCC1 architecture and catalytic activation.
]]></description>
<dc:creator>Jones, M. L.</dc:creator>
<dc:creator>Beuron, F.</dc:creator>
<dc:creator>Borg, A.</dc:creator>
<dc:creator>Nans, A.</dc:creator>
<dc:creator>Earl, C.</dc:creator>
<dc:creator>Briggs, D. C.</dc:creator>
<dc:creator>Bowles, M.</dc:creator>
<dc:creator>Morris, E. P.</dc:creator>
<dc:creator>Linch, M.</dc:creator>
<dc:creator>McDonald, N. Q.</dc:creator>
<dc:date>2019-10-07</dc:date>
<dc:identifier>doi:10.1101/796524</dc:identifier>
<dc:title><![CDATA[Cryo-EM Structures of the XPF-ERCC1 Endonuclease Reveal an Auto-Inhibited Conformation and the Basis for Activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/796151v1?rss=1">
<title>
<![CDATA[
Three-dimensional super-resolution fluorescence imaging of DNA 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/796151v1?rss=1"
</link>
<description><![CDATA[
Recent advances in fluorescence super-resolution microscopy are providing important insights into details of cellular structures. To acquire three dimensional (3D) super-resolution images of DNA, we combined binding activated localization microscopy (BALM) using fluorescent double-stranded DNA intercalators and optical astigmatism. We quantitatively establish the advantage of mono-over bis-intercalators before demonstrating the approach by visualizing single DNA molecules stretched between microspheres at various heights. Finally, the approach is applied to the more complex environment of intact and damaged metaphase chromosomes, unravelling their structural features.
]]></description>
<dc:creator>Yardimci, S.</dc:creator>
<dc:creator>Burnham, D. R.</dc:creator>
<dc:creator>Terry, S. Y. A.</dc:creator>
<dc:creator>Yardimci, H.</dc:creator>
<dc:date>2019-10-07</dc:date>
<dc:identifier>doi:10.1101/796151</dc:identifier>
<dc:title><![CDATA[Three-dimensional super-resolution fluorescence imaging of DNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/802363v1?rss=1">
<title>
<![CDATA[
Duplication and divergence of the retrovirus restriction gene Fv1 in Mus caroli mice allows protection from multiple retroviruses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/802363v1?rss=1"
</link>
<description><![CDATA[
Viruses and their hosts are locked in an evolutionary race where resistance to infection is acquired by the hosts while viruses develop strategies to circumvent these host defenses. Forming one arm of the host defense armory are cell autonomous restriction factors like Fv1. Originally described as protecting laboratory mice from infection by murine leukemia virus (MLV), Fv1s from some wild mice have also been found to restrict non-MLV retroviruses, suggesting an important role in the protection against viruses in nature. To begin to understand how restriction factors evolve, we surveyed the Fv1 genes of wild mice trapped in Thailand and characterized their restriction activities against a panel of retroviruses. An extra copy of the Fv1 gene, named Fv7, was found on chromosome 6 of three closely related Asian species of mice (Mus caroli, M. cervicolor and M. cookii). The presence of flanking repeats suggested it arose by LINE-mediated retrotransposition. A high degree of natural variation was observed in both Fv1 and Fv7, including numerous single nucleotide polymorphisms resulting in altered amino acids, as well as insertions and deletions that changed the length of the reading frames. These genes exhibited a range of restriction phenotypes with activities directed against feline foamy virus (FFV), equine infectious anemia virus (EIAV) and MLV. It seems likely, at least in the case of M. caroli, that the observed gene duplication confers protection against multiple viruses not possible with a single restriction factor. We suggest that EIAV-, FFV- and MLV-like viruses are endemic within these populations, driving the evolution of the Fv1 and Fv7 genes.nnAuthor SummaryDuring the passage of time all vertebrates will be exposed to infection by a variety of different kinds of virus. To meet this threat, a variety of genes for natural resistance to viral infection have evolved. The prototype of such so-called restriction factors is encoded by the mouse Fv1 gene, which acts to block the life cycle of retroviruses at a stage between virus entry into the cell and integration of the viral genetic material into the nuclear DNA. We have studied the evolution of this gene in certain species of wild mice from South East Asia and describe an example where a duplication of the Fv1 gene has taken place. The two copies of the gene, initially identical, have evolved separately allowing the development of resistance to two rather different kinds of retroviruses, lentiviruses and spumaviruses. Independent selection for resistance to these two kinds of retrovirus suggests that such mice are repeatedly exposed to never-before-reported pathogenic retroviruses of these genera.
]]></description>
<dc:creator>Yap, M. W.</dc:creator>
<dc:creator>Young, G. R.</dc:creator>
<dc:creator>Varnaite, R.</dc:creator>
<dc:creator>Morand, S.</dc:creator>
<dc:creator>Stoye, J. P.</dc:creator>
<dc:date>2019-10-11</dc:date>
<dc:identifier>doi:10.1101/802363</dc:identifier>
<dc:title><![CDATA[Duplication and divergence of the retrovirus restriction gene Fv1 in Mus caroli mice allows protection from multiple retroviruses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/809228v1?rss=1">
<title>
<![CDATA[
Salt-inducible kinases (SIKs) regulate TGFβ-mediated transcriptional and apoptotic responses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/809228v1?rss=1"
</link>
<description><![CDATA[
AbstractThe signalling pathways initiated by members of the transforming growth factor-{beta} (TGF{beta}) family of cytokines control many metazoan cellular processes, including proliferation and differentiation, epithelial-mesenchymal transition (EMT), and apoptosis. TGF{beta} signalling is therefore strictly regulated to ensure appropriate context-dependent physiological responses. In an attempt to identify novel regulatory components of the TGF{beta} signalling pathway, we performed a pharmacological screen using a cell line engineered to report the endogenous transcription of the TGF{beta}-responsive target gene PAI-1. The screen revealed that small-molecular inhibitors of salt-inducible kinases (SIKs) attenuate TGF{beta}-mediated transcription of PAI-1 without affecting receptor-mediated SMAD phosphorylation, SMAD complex formation or nuclear translocation. We provide evidence that genetic inactivation of SIK isoforms also attenuates TGF{beta}-dependent transcriptional responses. Pharmacological inhibition of SIKs using multiple small-molecule inhibitors potentiated apoptotic cell death induced by TGF{beta} stimulation. Our data therefore provides evidence for a novel function of SIKs in modulating TGF{beta}-mediated transcriptional and cellular responses.
]]></description>
<dc:creator>Hutchinson, L. D.</dc:creator>
<dc:creator>Darling, N. J.</dc:creator>
<dc:creator>Nicolaou, S.</dc:creator>
<dc:creator>Gori, I.</dc:creator>
<dc:creator>Squair, D. R.</dc:creator>
<dc:creator>Cohen, P.</dc:creator>
<dc:creator>Hill, C. S.</dc:creator>
<dc:creator>Sapkota, G. P.</dc:creator>
<dc:date>2019-10-17</dc:date>
<dc:identifier>doi:10.1101/809228</dc:identifier>
<dc:title><![CDATA[Salt-inducible kinases (SIKs) regulate TGFβ-mediated transcriptional and apoptotic responses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/821405v1?rss=1">
<title>
<![CDATA[
Divergent excitation two photon microscopy for 3D random access mesoscale imaging at single cell resolution 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/821405v1?rss=1"
</link>
<description><![CDATA[
ABSTACTIn neuroscience, diffraction limited two-photon (2P) microscopy is a cornerstone technique that permits minimally invasive optical monitoring of neuronal activity. However, most conventional 2P microscopes impose significant constraints on the size of the imaging field-of-view and the specific shape of the effective excitation volume, thus limiting the scope of biological questions that can be addressed and the information obtainable. Here, employing  divergent beam optics (DBO), we present an ultra-low-cost, easily implemented and flexible solution to address these limitations, offering a several-fold expanded three-dimensional field of view that also maintains single-cell resolution. We show that this implementation increases both the space-bandwidth product and effective excitation power, and allows for straight-forward tailoring of the point-spread-function. Moreover, rapid laser-focus control via an electrically tunable lens now allows near-simultaneous imaging of remote regions separated in three dimensions and permits the bending of imaging planes to follow natural curvatures in biological structures. Crucially, our core design is readily implemented (and reversed) within a matter of hours, and fully compatible with a wide range of existing 2P customizations, making it highly suitable as a base platform for further development. We demonstrate the application of our system for imaging neuronal activity in a variety of examples in mice, zebrafish and fruit flies.
]]></description>
<dc:creator>Janiak, F. K.</dc:creator>
<dc:creator>Bartel, P.</dc:creator>
<dc:creator>Bale, M.</dc:creator>
<dc:creator>T, Y.</dc:creator>
<dc:creator>Komulainen, E. H.</dc:creator>
<dc:creator>Zhou, M.</dc:creator>
<dc:creator>Staras, K.</dc:creator>
<dc:creator>Prieto-Godino, L. L.</dc:creator>
<dc:creator>Euler, T.</dc:creator>
<dc:creator>Maravall, M.</dc:creator>
<dc:creator>Baden, T.</dc:creator>
<dc:date>2019-10-29</dc:date>
<dc:identifier>doi:10.1101/821405</dc:identifier>
<dc:title><![CDATA[Divergent excitation two photon microscopy for 3D random access mesoscale imaging at single cell resolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/820076v1?rss=1">
<title>
<![CDATA[
Mimicry embedding for advanced neuralnetwork training of 3D biomedical micrographs 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/820076v1?rss=1"
</link>
<description><![CDATA[
The use of deep neural networks (DNNs) for analysis of complex biomedical images shows great promise but is hampered by a lack of large verified datasets for rapid network evolution. Here we present a novel "mimicry embedding" strategy for rapid application of neural network architecture-based analysis of biomedical imaging datasets. Embedding of a novel biological dataset, such that it mimics a verified dataset, enables efficient deep learning and seamless architecture switching. We apply this strategy across various microbiological phenotypes; from super-resolved viruses to in vivo parasitic infections. We demonstrate that mimicry embedding enables efficient and accurate analysis of three-dimensional microscopy datasets. The results suggest that transfer learning from pre-trained network data may be a powerful general strategy for analysis of heterogeneous biomedical imaging datasets.
]]></description>
<dc:creator>Yakimovich, A.</dc:creator>
<dc:creator>Huttunen, M.</dc:creator>
<dc:creator>Samolej, J.</dc:creator>
<dc:creator>Clough, B.</dc:creator>
<dc:creator>Yoshida, N.</dc:creator>
<dc:creator>Mostowy, S.</dc:creator>
<dc:creator>Frickel, E.</dc:creator>
<dc:creator>Mercer, J.</dc:creator>
<dc:date>2019-10-28</dc:date>
<dc:identifier>doi:10.1101/820076</dc:identifier>
<dc:title><![CDATA[Mimicry embedding for advanced neuralnetwork training of 3D biomedical micrographs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/828624v1?rss=1">
<title>
<![CDATA[
In vivo control of Toxoplasma gondii by zebrafish macrophages 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/828624v1?rss=1"
</link>
<description><![CDATA[
Toxoplasma gondii is an obligate intracellular parasite capable of invading any nucleated cell. Three main clonal lineages (type I, II, III) exist and murine models have driven the understanding of general and strain-specific immune mechanisms underlying Toxoplasma infection. However, murine models are limited for studying parasite-leukocyte interactions in vivo, and discrepancies exist between cellular immune responses observed in mouse versus human cells. Here, we develop a zebrafish infection model to study the innate immune response to Toxoplasma in vivo. By infecting the zebrafish hindbrain ventricle, and using high-resolution microscopy techniques coupled with computer vision driven automated image analysis, we reveal that Toxoplasma invades and replicates inside a parasitophorous vacuole to which type I and III parasites recruit host cell mitochondria. We show that type II and III strains maintain a higher infectious burden than type I strains. To understand how parasites are being cleared in vivo, we analyzed Toxoplasma-macrophage interactions using time-lapse and correlative light and electron microscopy. Strikingly, macrophages are recruited to the infection site and play a key role in Toxoplasma control. These results highlight in vivo control of Toxoplasma by macrophages, and illuminate the possibility to exploit zebrafish for discoveries within the field of parasite immunity.
]]></description>
<dc:creator>Yoshida, N.</dc:creator>
<dc:creator>Domart, M.-C.</dc:creator>
<dc:creator>Yakimovich, A.</dc:creator>
<dc:creator>Mazon-Moya, M. J.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:creator>Mercer, J.</dc:creator>
<dc:creator>Frickel, E.</dc:creator>
<dc:creator>Mostowy, S.</dc:creator>
<dc:date>2019-11-02</dc:date>
<dc:identifier>doi:10.1101/828624</dc:identifier>
<dc:title><![CDATA[In vivo control of Toxoplasma gondii by zebrafish macrophages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/829739v1?rss=1">
<title>
<![CDATA[
Subcellular mRNA localization regulates ribosome biogenesis in migrating cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/829739v1?rss=1"
</link>
<description><![CDATA[
Translation of Ribosomal Protein-coding mRNAs (RP-mRNAs) constitutes a key step in ribosome biogenesis, but the mechanisms which modulate RP-mRNAs translation in coordination with other cellular processes are poorly defined. Here we show that the subcellular localization of RP-mRNAs acts as a key regulator of their translation during cell migration. As cells migrate into their surroundings, RP-mRNAs localize to actin-rich protrusions at the front the cells. This localization is mediated by La-related protein 6 (LARP6), an RNA binding protein that is enriched in protrusions. Protrusions act as hotspots of translation for RP-mRNAs, resulting in enhancement of ribosome biogenesis and overall protein synthesis, which is required for sustained migration. In human breast carcinomas, Epithelial to Mesenchymal Transition (EMT) upregulates LARP6 expression to enhance ribosome biogenesis and support invasive growth. Our findings reveal LARP6 mediated mRNA localization as a key regulator of ribosome biogenesis during cell migration, and demonstrate a role for this process in cancer progression downstream of EMT.
]]></description>
<dc:creator>Dermit, M.</dc:creator>
<dc:creator>Dodel, M.</dc:creator>
<dc:creator>Lee, F. C. Y.</dc:creator>
<dc:creator>Azman, M. S.</dc:creator>
<dc:creator>Schwenzer, H.</dc:creator>
<dc:creator>Jones, J. L.</dc:creator>
<dc:creator>Blagden, S. P.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Mardakheh, F. K.</dc:creator>
<dc:date>2019-11-04</dc:date>
<dc:identifier>doi:10.1101/829739</dc:identifier>
<dc:title><![CDATA[Subcellular mRNA localization regulates ribosome biogenesis in migrating cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/823716v1?rss=1">
<title>
<![CDATA[
Escape from nonsense mediated decay associates with anti-tumor immunogenicity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/823716v1?rss=1"
</link>
<description><![CDATA[
Frameshift insertion/deletions (fs-indels) are an infrequent but potentially highly immunogenic mutation subtype. Although fs-indel transcripts are susceptible to degradation through the non-sense mediated decay (NMD) pathway, we hypothesise that some fs-indels escape degradation and lead to an increased abundance of tumor specific neoantigens, that are highly distinct from self. We analysed matched DNA and RNA sequencing data from TCGA, and five separate melanoma cohorts treated with immunotherapy. Using allele-specific expression analysis we show that expressed fs-indels were enriched in genomic positions predicted to escape NMD, and associated with higher protein expression, consistent with degradation escape ("NMD-escape"). Across four independent cohorts, fs-indel NMD-escape mutations were found to be significantly associated with clinical benefit to checkpoint inhibitor (CPI) therapy (Pmeta=0.0039), a stronger association than either nsSNV (Pmeta=0.073) or fs-indel (Pmeta=0.064) count. NMD-escape mutations were additionally shown to have independent predictive power in the "low-TMB" setting, and may serve as a biomarker to rescue patients judged ineligible for CPI based on overall TMB, but still with a high chance of response (low-TMB cohort: NMD-escape-positive % clinical benefit=53%, NMD-escape-negative % clinical benefit=16%, P=0.0098). Furthermore, in an adoptive cell therapy (ACT) treated cohort, NMD-escape mutation count was the most significant biomarker associated with clinical benefit (P=0.021). Analysis of functional T-cell reactivity screens from recent personalized vaccine and CPI studies shows direct evidence of fs-indel derived neoantigens eliciting patient anti-tumor immune response (n=15). We additionally observe a subset of fs-indel mutations, with highly elongated neo open reading frames, which are found to be significantly enriched for immunogenic reactivity in these patient studies (P=0.0032). Finally, consistent with the potency of NMD-escape derived neo-antigens and ongoing immune-editing, NMD-escape fs-indels appear to be under negative selective pressure in untreated TCGA cases. Given the strongly immunogenic potential, and relatively rare nature of NMD-escape fs-indels, these alterations may be attractive candidates in immunotherapy biomarker optimisation and neoantigen ACT or vaccine strategies.
]]></description>
<dc:creator>Litchfield, K.</dc:creator>
<dc:creator>Reading, J.</dc:creator>
<dc:creator>Lim, E.</dc:creator>
<dc:creator>Xu, H.</dc:creator>
<dc:creator>Liu, P.</dc:creator>
<dc:creator>Al-Bakir, M.</dc:creator>
<dc:creator>Wong, S.</dc:creator>
<dc:creator>Rowan, A.</dc:creator>
<dc:creator>Funt, S.</dc:creator>
<dc:creator>Merghoub, T.</dc:creator>
<dc:creator>Lauss, M.</dc:creator>
<dc:creator>Svane, I. M.</dc:creator>
<dc:creator>Jonsson, G.</dc:creator>
<dc:creator>Herrero, J.</dc:creator>
<dc:creator>Larkin, J.</dc:creator>
<dc:creator>Quezada, S. A.</dc:creator>
<dc:creator>Hellmann, M. D.</dc:creator>
<dc:creator>Turajlic, S.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:date>2019-11-07</dc:date>
<dc:identifier>doi:10.1101/823716</dc:identifier>
<dc:title><![CDATA[Escape from nonsense mediated decay associates with anti-tumor immunogenicity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/836114v1?rss=1">
<title>
<![CDATA[
Functional characterisation of gut microbiota and metabolism in Type 2 diabetes indicates that Clostridiales and Enterococcus could play a key role in the disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/836114v1?rss=1"
</link>
<description><![CDATA[
There is growing evidence indicating that gut microbiota contributes to the development of metabolic syndrome and Type 2 Diabetes (T2D). The most widely-used model for T2D research is the leptin deficient db/db mouse model. Yet, a characterisation of the gut microbial composition in this model in relationship with the metabolism is lacking. The objectives of this study were to identify metabolomics and microbial modulations associated with T2D in the db/db mouse model. The majority of microbial changes observed included an increase of Enterobacteriaceae and a decrease of Clostridiales in diabetics. The metabolomics interrogation of caecum indicated a lower proteolytic activity in diabetics, who also showed higher Short-Chain Fatty Acid (SCFA) levels. In the case of faeces, the model identified 9 metabolites, the main ones were acetate, butyrate and Branched Chain Amino Acids (BCAAs). Finally, liver was the organ with more metabolic links with gut-microbiota followed by the Gut-Brain Axis (GBA). In conclusion, the interaction between Clostridiales and Enterococcus with caecal metabolism could play a key role in the onset and development of diabetes. Further studies should investigate whether the role of these bacteria is causal or co-occurring.
]]></description>
<dc:creator>Mora-Ortiz, M.</dc:creator>
<dc:creator>Oregioni, A.</dc:creator>
<dc:creator>Claus, S. P.</dc:creator>
<dc:date>2019-11-09</dc:date>
<dc:identifier>doi:10.1101/836114</dc:identifier>
<dc:title><![CDATA[Functional characterisation of gut microbiota and metabolism in Type 2 diabetes indicates that Clostridiales and Enterococcus could play a key role in the disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/833004v1?rss=1">
<title>
<![CDATA[
Immune surveillance in clinical regression of pre-invasive squamous cell lung cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/833004v1?rss=1"
</link>
<description><![CDATA[
Before squamous cell lung cancer develops, pre-cancerous lesions can be found in the airways. From longitudinal monitoring, we know that only half of such lesions become cancer, whereas a third spontaneously regress. While recent studies have described the presence of an active immune response in high-grade lesions, the mechanisms underpinning clinical regression of pre-cancerous lesions remain unknown. Here, we show that host immune surveillance is strongly implicated in lesion regression. Using bronchoscopic biopsies from human subjects, we find that regressive carcinoma in-situ lesions harbour more infiltrating immune cells than those that progress to cancer. Moreover, molecular profiling of these lesions identifies potential immune escape mechanisms specifically in those that progress to cancer: antigen presentation is impaired by genomic and epigenetic changes, TGF-beta signalling is overactive, and the immunomodulator TNFSF9 is downregulated. Changes appear intrinsic to the CIS lesions as the adjacent stroma of progressive and regressive lesions are transcriptomically similar. This study identifies mechanisms by which pre-cancerous lesions evade immune detection during the earliest stages of carcinogenesis and forms a basis for new therapeutic strategies that treat or prevent early stage lung cancer.
]]></description>
<dc:creator>Pennycuick, A.</dc:creator>
<dc:creator>Teixeira, V. H.</dc:creator>
<dc:creator>AbdulJabbar, K.</dc:creator>
<dc:creator>Raza, S. E. A.</dc:creator>
<dc:creator>Lund, T.</dc:creator>
<dc:creator>Akarca, A.</dc:creator>
<dc:creator>Rosenthal, R.</dc:creator>
<dc:creator>Pipinikas, C. P.</dc:creator>
<dc:creator>Lee-Six, H.</dc:creator>
<dc:creator>Chandrasekharan, D. P.</dc:creator>
<dc:creator>Hynds, R. E.</dc:creator>
<dc:creator>Gowers, K. H. C.</dc:creator>
<dc:creator>Henry, J. Y.</dc:creator>
<dc:creator>Denais, C.</dc:creator>
<dc:creator>Falzon, M.</dc:creator>
<dc:creator>Antoniou, S.</dc:creator>
<dc:creator>Durrenberger, P. F.</dc:creator>
<dc:creator>Furness, A.</dc:creator>
<dc:creator>Carroll, B.</dc:creator>
<dc:creator>Thakrar, R. M.</dc:creator>
<dc:creator>George, P. J.</dc:creator>
<dc:creator>Marafioti, T.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Thirlwell, C.</dc:creator>
<dc:creator>Campbell, P. J.</dc:creator>
<dc:creator>Yuan, Y.</dc:creator>
<dc:creator>Quezada, S. A.</dc:creator>
<dc:creator>McGranahan, N.</dc:creator>
<dc:creator>Janes, S. M.</dc:creator>
<dc:date>2019-11-10</dc:date>
<dc:identifier>doi:10.1101/833004</dc:identifier>
<dc:title><![CDATA[Immune surveillance in clinical regression of pre-invasive squamous cell lung cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/843136v1?rss=1">
<title>
<![CDATA[
RNA modifications detection by comparative Nanopore direct RNA sequencing 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/843136v1?rss=1"
</link>
<description><![CDATA[
RNA molecules undergo a vast array of chemical post-transcriptional modifications (PTMs) that can affect their structure and interaction properties. To date, over 150 naturally occurring PTMs have been identified, however the overwhelming majority of their functions remain elusive. In recent years, a small number of PTMs have been successfully mapped to the transcriptome using experimental approaches relying on high-throughput sequencing. Oxford Nanopore direct-RNA sequencing (DRS) technology has been shown to be sensitive to RNA modifications. We developed and validated Nanocompore, a robust analytical framework to evaluate the presence of modifications in DRS data. To do so, we compare an RNA sample of interest against a non-modified control sample. Our strategy does not require a training set and allows the use of replicates to model biological variability. Here, we demonstrate the ability of Nanocompore to detect RNA modifications at single-molecule resolution in human polyA+ RNAs, as well as in targeted non-coding RNAs. Our results correlate well with orthogonal methods, confirm previous observations on the distribution of N6-methyladenosine sites and provide novel insights into the distribution of RNA modifications in the coding and non-coding transcriptomes. The latest version of Nanocompore can be obtained at https://github.com/tleonardi/nanocompore.
]]></description>
<dc:creator>Leger, A.</dc:creator>
<dc:creator>Amaral, P. P.</dc:creator>
<dc:creator>Pandolfini, L.</dc:creator>
<dc:creator>Capitanchik, C.</dc:creator>
<dc:creator>Capraro, F.</dc:creator>
<dc:creator>Barbieri, I.</dc:creator>
<dc:creator>Migliori, V.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:creator>Enright, A. J.</dc:creator>
<dc:creator>Tzelepis, K.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Fitzgerald, T.</dc:creator>
<dc:creator>Birney, E.</dc:creator>
<dc:creator>Leonardi, T.</dc:creator>
<dc:creator>Kouzarides, T.</dc:creator>
<dc:date>2019-11-15</dc:date>
<dc:identifier>doi:10.1101/843136</dc:identifier>
<dc:title><![CDATA[RNA modifications detection by comparative Nanopore direct RNA sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/843458v1?rss=1">
<title>
<![CDATA[
Integrated Genomic Analysis Reveals Key Features of Long Undecoded Transcript Isoform (LUTI)-based Gene Repression 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/843458v1?rss=1"
</link>
<description><![CDATA[
Long Undecoded Transcript Isoforms (LUTIs) represent a class of non-canonical mRNAs that downregulate gene expression through the combined act of transcriptional and translational repression. While single gene studies revealed some important aspects of LUTI-based repression, how these features impact gene regulation at a global scale is unknown. By using transcript leader and direct RNA sequencing, here we identify 74 LUTI candidates that are expressed specifically during meiotic prophase. Translational repression of these candidates is ubiquitous and dependent on upstream open reading frames. However, LUTI-based transcriptional repression is highly variable. In only 50% of the cases, LUTI transcription causes downregulation of the protein-coding transcript isoform. Higher LUTI expression, enrichment of histone 3 lysine 36 trimethylation, and changes in nucleosome position are the strongest predictors of LUTI-based transcriptional repression. We conclude that LUTIs downregulate gene expression in a manner that integrates translational repression, chromatin state changes, and the magnitude of LUTI expression.
]]></description>
<dc:creator>Tresenrider, A.</dc:creator>
<dc:creator>Jorgensen, V.</dc:creator>
<dc:creator>Chia, M.</dc:creator>
<dc:creator>Liao, H.</dc:creator>
<dc:creator>van Werven, F.</dc:creator>
<dc:creator>Unal, E.</dc:creator>
<dc:date>2019-11-15</dc:date>
<dc:identifier>doi:10.1101/843458</dc:identifier>
<dc:title><![CDATA[Integrated Genomic Analysis Reveals Key Features of Long Undecoded Transcript Isoform (LUTI)-based Gene Repression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/848580v1?rss=1">
<title>
<![CDATA[
FUS gene is dual-coding with both proteins united in molecular hallmarks of amyotrophic lateral sclerosis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/848580v1?rss=1"
</link>
<description><![CDATA[
Novel functional coding sequences (altORFs) are camouflaged within annotated ones (CDS) in a different reading frame. We discovered an altORF nested in the FUS CDS encoding a conserved 169 amino acid protein, altFUS. AltFUS is endogenously expressed in human tissues, notably in the motor cortex and motor neurons. Overexpression of wild-type FUS and/or amyotrophic lateral sclerosis-linked FUS mutants is known to trigger toxic mechanisms in different models. These include an inhibition of autophagy, loss of mitochondrial potential, and accumulation of cytoplasmic aggregates. We show here that altFUS, not FUS, is responsible for the inhibition of autophagy. AltFUS is also pivotal in the mechanisms leading to the mitochondrial potential loss and accumulation of cytoplasmic aggregates. Suppression of altFUS expression in a Drosophila model of FUS-related toxicity protects against neurodegeneration. Some mutations found in ALS patients are overlooked because of their synonymous effect on the FUS protein. Yet we showed they exert a deleterious effect via their missense consequence on the overlapping altFUS protein. These findings demonstrate that FUS is a bicistronic gene and suggest that both proteins, FUS and altFUS, cooperate in toxic mechanisms.
]]></description>
<dc:creator>Brunet, M. A.</dc:creator>
<dc:creator>Jacques, J.-F.</dc:creator>
<dc:creator>Nassari, S.</dc:creator>
<dc:creator>Tyzack, G. E.</dc:creator>
<dc:creator>McGoldrick, P.</dc:creator>
<dc:creator>Zinman, L.</dc:creator>
<dc:creator>Jean, S.</dc:creator>
<dc:creator>Robertson, J.</dc:creator>
<dc:creator>Patani, R.</dc:creator>
<dc:creator>Roucou, X.</dc:creator>
<dc:date>2019-11-20</dc:date>
<dc:identifier>doi:10.1101/848580</dc:identifier>
<dc:title><![CDATA[FUS gene is dual-coding with both proteins united in molecular hallmarks of amyotrophic lateral sclerosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/850321v1?rss=1">
<title>
<![CDATA[
Cyclic peptides can engage a single binding pocket through multiple, entirely divergent modes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/850321v1?rss=1"
</link>
<description><![CDATA[
Cyclic peptide display screening techniques can identify drug leads and biological probes with exceptional affinity and specificity. To date, however, the structural and functional diversity encoded in such peptide libraries remains unexplored. We have used the Random nonstandard Peptide Integrated Discovery (RaPID) system to develop cyclic peptide inhibitors of several acetyllysine-binding bromodomains from the Bromodomain and Extra-Terminal domain (BET) family of epigenetic regulators. These peptides have very high affinities for their targets and exhibit extraordinary selectivity (up to 106-fold), making them the highest-affinity and most specific BET-binding molecules discovered to date. Crystal structures of 13 distinct peptide-bromodomain complexes, which all target the acetyllysine-binding pocket, reveal remarkable diversity in both peptide structure and binding mode, and include both -helical and {beta}-sheet type structures. The peptides can exhibit a high degree of structural pre-organization and bivalent binding of two BDs by one peptide was common, flagging the potential for a new direction in inhibitor design that could bring stronger discrimination between BET-family paralogues. Our data demonstrate for the first time the enormous potential held in these libraries to provide a wide array of modes against a single target, maximizing the opportunity to attain high potency and specificity ligands to a wide variety of proteins.
]]></description>
<dc:creator>Patel, K.</dc:creator>
<dc:creator>Walport, L. J.</dc:creator>
<dc:creator>Walshe, J. L.</dc:creator>
<dc:creator>Solomon, P.</dc:creator>
<dc:creator>Low, J. K. K.</dc:creator>
<dc:creator>Tran, D. H.</dc:creator>
<dc:creator>Mouradian, K. S.</dc:creator>
<dc:creator>Silva, A. P. G.</dc:creator>
<dc:creator>Wilkinson-White, L.</dc:creator>
<dc:creator>Matthews, J. M.</dc:creator>
<dc:creator>Guss, J. M.</dc:creator>
<dc:creator>Payne, R. J.</dc:creator>
<dc:creator>Passioura, T.</dc:creator>
<dc:creator>Suga, H.</dc:creator>
<dc:creator>Mackay, J. P.</dc:creator>
<dc:date>2019-11-21</dc:date>
<dc:identifier>doi:10.1101/850321</dc:identifier>
<dc:title><![CDATA[Cyclic peptides can engage a single binding pocket through multiple, entirely divergent modes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/853218v1?rss=1">
<title>
<![CDATA[
Microtubule Nucleation by Single Human γTuRC in a Partly Open Asymmetric Conformation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/853218v1?rss=1"
</link>
<description><![CDATA[
The {gamma}-tubulin ring complex ({gamma}TuRC) is the major microtubule nucleator in cells. However, the mechanism of its regulation is not understood. Here, we purified human {gamma}TuRC and quantitatively characterized its nucleation properties in a TIRF microscopy-based real-time nucleation assay. We find that microtubule nucleation by {gamma}TuRC is kinetically inhibited compared to microtubule elongation. Determining the cryo-EM structure of {gamma}TuRC at 4 [A] resolution reveals an asymmetric conformation with only part of the complex in a  closed conformation matching the microtubule geometry. Several factors stabilise the closed conformation. One is actin in the core of the complex and others, likely MZT1 or MZT2, line the outer perimeter of the closed part of {gamma}TuRC. The opposed side of {gamma}TuRC is in an  open, nucleation-incompetent conformation, leading to a structural asymmetry, explaining the kinetic inhibition of nucleation by human {gamma}TuRC. Our data suggest possible regulatory mechanisms for microtubule nucleation by {gamma}TuRC closure.
]]></description>
<dc:creator>Consolati, T.</dc:creator>
<dc:creator>Locke, J.</dc:creator>
<dc:creator>Roostalu, J.</dc:creator>
<dc:creator>Asthana, J.</dc:creator>
<dc:creator>Lim, W. M.</dc:creator>
<dc:creator>Gannon, J.</dc:creator>
<dc:creator>Martino, F.</dc:creator>
<dc:creator>Costa, A.</dc:creator>
<dc:creator>Surrey, T.</dc:creator>
<dc:date>2019-11-23</dc:date>
<dc:identifier>doi:10.1101/853218</dc:identifier>
<dc:title><![CDATA[Microtubule Nucleation by Single Human γTuRC in a Partly Open Asymmetric Conformation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/854083v1?rss=1">
<title>
<![CDATA[
Engineering transplantable jejunal mucosal grafts using primary patient-derived organoids from children with intestinal failure 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/854083v1?rss=1"
</link>
<description><![CDATA[
Intestinal failure (IF), following extensive anatomical or functional loss of small intestine (SI), has debilitating long-term effects on infants and children with this condition. Priority of care is to increase the childs length of functional intestine, jejunum in particular, to improve nutritional independence. Here we report a robust protocol for reconstruction of autologous intestinal mucosal grafts using primary IF patient materials. Human jejunal intestinal organoids derived from paediatric IF patients can be expanded efficiently in vitro with region-specific markers preserved after long-term culture. Decellularized human intestinal matrix with intact ultrastructure is used as biological scaffolds. Proteomic and Raman spectroscopic analyses reveal highly analogous biochemical composition of decellularized human SI and colon matrix, implying that they can both be utilised as scaffolds for jejunal graft reconstruction. Indeed, seeding of primary human jejunal organoids to either SI or colonic scaffolds in vitro can efficiently reconstruct functional jejunal grafts with persistent disaccharidase activity as early as 4 days after seeding, which can further survive and mature after transplantation in vivo. Our findings pave the way towards regenerative medicine for IF patients.
]]></description>
<dc:creator>Meran, L.</dc:creator>
<dc:creator>Massie, I.</dc:creator>
<dc:creator>Weston, A.</dc:creator>
<dc:creator>Gaifulina, R.</dc:creator>
<dc:creator>Faull, P.</dc:creator>
<dc:creator>Orford, M.</dc:creator>
<dc:creator>Kucharska, A.</dc:creator>
<dc:creator>Baulies, A.</dc:creator>
<dc:creator>Hirst, E.</dc:creator>
<dc:creator>Konig, J.</dc:creator>
<dc:creator>Tedeschi, A. M.</dc:creator>
<dc:creator>Pellegata, A. F.</dc:creator>
<dc:creator>Eli, S.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:creator>Thapar, N.</dc:creator>
<dc:creator>Thomas, G.</dc:creator>
<dc:creator>Eaton, S.</dc:creator>
<dc:creator>Bonfanti, P.</dc:creator>
<dc:creator>De Coppi, P.</dc:creator>
<dc:creator>Li, V. S. W.</dc:creator>
<dc:date>2019-11-25</dc:date>
<dc:identifier>doi:10.1101/854083</dc:identifier>
<dc:title><![CDATA[Engineering transplantable jejunal mucosal grafts using primary patient-derived organoids from children with intestinal failure]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/855361v1?rss=1">
<title>
<![CDATA[
The breast cancer oncogene IKKε coordinates mitochondrial function and serine metabolism 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/855361v1?rss=1"
</link>
<description><![CDATA[
The I{kappa}B kinase {varepsilon} (IKK{varepsilon}) is a key molecule at the crossroads of inflammation and cancer. Known for its role as an activator of NF{kappa}B and IRF3 signalling leading to cytokine secretion, the kinase is also a breast cancer oncogene, overexpressed in a variety of tumours. However, to what extent IKK{varepsilon} remodels cellular metabolism is currently unknown. Here we used a combination of metabolomics and phosphoproteomics to show that IKK{varepsilon} orchestrates a complex metabolic reprogramming that affects mitochondrial metabolism and serine biosynthesis. Acting independently of its canonical signalling role, IKK{varepsilon} upregulates the serine biosynthesis pathway (SBP) mainly by limiting glucose and pyruvate derived anaplerosis of the TCA cycle. In turn, this elicits activation of the transcription factor ATF4 and upregulation of the SBP genes. Importantly, pharmacological inhibition of the IKK{varepsilon}-induced metabolic phenotype reduces proliferation of breast cancer cells. Finally, we show that in a set of basal ER negative and highly proliferative human breast cancer tumours, IKK{varepsilon} and PSAT1 expression levels are positively correlated corroborating the link between IKK{varepsilon} and the SBP in the clinical context.
]]></description>
<dc:creator>Xu, R.</dc:creator>
<dc:creator>Jones, W.</dc:creator>
<dc:creator>Wilcz-Villega, E.</dc:creator>
<dc:creator>Costa, S.</dc:creator>
<dc:creator>Rajeeve, V.</dc:creator>
<dc:creator>Bentham, R. B.</dc:creator>
<dc:creator>Bryson, K.</dc:creator>
<dc:creator>Nagano, A.</dc:creator>
<dc:creator>Yaman, B.</dc:creator>
<dc:creator>Olendo Barasa, S.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Chelala, C.</dc:creator>
<dc:creator>Cutillas, P.</dc:creator>
<dc:creator>Szabadkai, G.</dc:creator>
<dc:creator>Frezza, C.</dc:creator>
<dc:creator>Bianchi, K.</dc:creator>
<dc:date>2019-11-26</dc:date>
<dc:identifier>doi:10.1101/855361</dc:identifier>
<dc:title><![CDATA[The breast cancer oncogene IKKε coordinates mitochondrial function and serine metabolism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/862391v1?rss=1">
<title>
<![CDATA[
Cryogenic OrbiSIMS Localizes Semi-Volatile Molecules in Biological Tissues 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/862391v1?rss=1"
</link>
<description><![CDATA[
OrbiSIMS is a recently developed instrument for label-free imaging of chemicals with micron spatial resolution and high mass resolution. Here we report a cryogenic workflow for OrbiSIMS (Cryo-OrbiSIMS) that improves chemical detection of lipids and other biomolecules in tissues. Cryo-OrbiSIMS decreases ion-beam induced fragmentation, allowing large molecules such as triglycerides to be more reliably identified. It also increases chemical coverage to include biomolecules with intermediate or high vapor pressures, such as free fatty acids and semi-volatile organic compounds (SVOCs). We find that Cryo-OrbiSIMS reveals the hitherto unknown localization patterns of SVOCs with high spatial and chemical resolution in diverse plant, animal and human tissues. We also show that Cryo-OrbiSIMS can be combined with genetic analysis to identify enzymes regulating SVOC metabolism. Cryo-OrbiSIMS is applicable to high resolution imaging of a wide variety of non-volatile and semi-volatile molecules across many areas of biomedicine.
]]></description>
<dc:creator>Newell, C. L.</dc:creator>
<dc:creator>Vorng, J.-L.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Gilmore, I.</dc:creator>
<dc:creator>Gould, A. P.</dc:creator>
<dc:date>2019-12-02</dc:date>
<dc:identifier>doi:10.1101/862391</dc:identifier>
<dc:title><![CDATA[Cryogenic OrbiSIMS Localizes Semi-Volatile Molecules in Biological Tissues]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/863548v1?rss=1">
<title>
<![CDATA[
The apparent requirement for protein synthesis during G2 phase is due to checkpoint activation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/863548v1?rss=1"
</link>
<description><![CDATA[
Protein synthesis inhibitors (e.g. cycloheximide) prevent cells from entering mitosis, suggesting that cell cycle progression requires protein synthesis until right before mitotic entry. However, cycloheximide is also known to activate p38 MAPK, which can delay mitotic entry through a G2/M checkpoint. Here we asked whether checkpoint activation or a requirement for protein synthesis is responsible for the cycloheximide effect. We found that p38 inhibitors prevent cycloheximide-treated cells from arresting in G2 phase, and that G2 duration is normal in about half of these cells. The Wee1/Myt1 inhibitor PD0166285 also prevents cycloheximide from blocking mitotic entry, raising the possibility that Wee1 and/or Myt1 mediate the cycloheximide-induced G2 arrest. Thus, the ultimate trigger for mitotic entry appears not to be the continued synthesis of mitotic cyclins or other proteins. However, M-phase progression was delayed in cycloheximide-plus-kinase-inhibitor-treated cells, emphasizing the different requirements of protein synthesis for timely entry and completion of mitosis.

Impact statementCycloheximide arrests cells in G2 phase due to activation of p38 MAPK, not inhibition of protein synthesis, arguing that protein synthesis in G2 phase is not required for mitotic entry.
]]></description>
<dc:creator>Lockhead, S.</dc:creator>
<dc:creator>Moskaleva, A.</dc:creator>
<dc:creator>Kamenz, J.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Kang, M.</dc:creator>
<dc:creator>Reddy, A. R. K.</dc:creator>
<dc:creator>Santos, S.</dc:creator>
<dc:creator>Ferrell, J. E.</dc:creator>
<dc:date>2019-12-03</dc:date>
<dc:identifier>doi:10.1101/863548</dc:identifier>
<dc:title><![CDATA[The apparent requirement for protein synthesis during G2 phase is due to checkpoint activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/865725v1?rss=1">
<title>
<![CDATA[
Widespread use of the "ascidian" mitochondrial genetic code in tunicates 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/865725v1?rss=1"
</link>
<description><![CDATA[
BackgroundAscidians, a tunicate class, use a mitochondrial genetic code that is distinct from vertebrates and other invertebrates. Though it has been used to translate the coding sequences from other tunicate species on a case-by-case basis, it is has not been investigated whether this can be done systematically. This is an important because a) some tunicate mitochondrial sequences are currently translated with the invertebrate code by repositories such as NCBIs GenBank, and b) uncertainties about the genetic code to use can complicate or introduce errors in phylogenetic studies based on translated mitochondrial protein sequences.

MethodsWe collected publicly available nucleotide sequences for non-ascidian tunicates including appendicularians such as Oikopleura dioica, translated them using the ascidian mitochondrial code, and built multiple sequence alignments covering all tunicate classes.

ResultsAll tunicates studied here appear to translate AGR codons to glycine instead of serine (invertebrates) or as a stop codon (vertebrates), as initially described in ascidians. Among Oikopleuridae, we suggest further possible changes in the use of the ATA (Ile [-&gt;] Met) and TGA (Trp [-&gt;] Arg) codons.

ConclusionsWe recommend using the ascidian mitochondrial code in automatic translation pipelines of mitochondrial sequences for all tunicates. Further investigation is required for additional species-specific differences.
]]></description>
<dc:creator>Pichon, J.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:creator>Plessy, C.</dc:creator>
<dc:date>2019-12-05</dc:date>
<dc:identifier>doi:10.1101/865725</dc:identifier>
<dc:title><![CDATA[Widespread use of the "ascidian" mitochondrial genetic code in tunicates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/867507v1?rss=1">
<title>
<![CDATA[
A FIJI Macro for quantifying pattern in extracellular matrix 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/867507v1?rss=1"
</link>
<description><![CDATA[
Diverse extracellular matrix patterns are observed in both normal and pathological tissue. However, most current tools for quantitative analysis focus on a single aspect of matrix patterning. Thus, an automated pipeline that simultaneously quantifies a broad range of metrics and enables a comprehensive description of varied matrix patterns is needed. To this end we have developed an ImageJ plugin called TWOMBLI, which stands for The Workflow Of Matrix BioLogy Informatics. TWOMBLI is designed to be quick, versatile and easy-to-use particularly for non-computational scientists. TWOMBLI can be downloaded from https://github.com/wershofe/TWOMBLI together with detailed documentation. Here we present an overview of the pipeline together with examples from a wide range of contexts where matrix patterns are generated.
]]></description>
<dc:creator>Wershof, E.</dc:creator>
<dc:creator>Barry, D. J.</dc:creator>
<dc:creator>Jenkins, R. P.</dc:creator>
<dc:creator>Rullan, A.</dc:creator>
<dc:creator>Wilkins, A.</dc:creator>
<dc:creator>Roxanis, I.</dc:creator>
<dc:creator>Anderson, K. I.</dc:creator>
<dc:creator>Park, D.</dc:creator>
<dc:creator>Bates, P. A.</dc:creator>
<dc:creator>Sahai, E.</dc:creator>
<dc:date>2019-12-06</dc:date>
<dc:identifier>doi:10.1101/867507</dc:identifier>
<dc:title><![CDATA[A FIJI Macro for quantifying pattern in extracellular matrix]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/872846v1?rss=1">
<title>
<![CDATA[
Role of SHIP2 in cell repulsion regulated by Eph receptor and ephrin signaling 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/872846v1?rss=1"
</link>
<description><![CDATA[
Previous studies have found that activation of EphB2 and ephrinB1 that drives cell segregation leads to phosphorylation of the phosphoinositide phosphatase SHIP2 downstream of forward (EphB2) but not reverse (ephrinB1) signaling. We have analysed whether SHIP2 interacts with EphB2 and contributes to cell responses to EphB2-ephrinB1 signaling. We confirm that EphB2 activation leads to SHIP2 phosphorylation on Y1135 and find that they interact through the SH2 domain of SHIP2. There is thus a distinct mode of interaction from EphA2, which binds SHIP2 via its SAM domain. Knockdown of SHIP2 in EphB2 cells leads to decreased segregation from ephrinB1 cells, and a decrease in the repulsion response of EphB2 cells. SHIP2 knockdown in ephrinB1 cells also decreases their repulsion response, but does not disrupt segregation which is largely driven by forward signaling. These findings show that activation of EphB2 leads to recruitment and phosphorylation of SHIP2, and that SHIP2 contributes to cell repulsion responses that underlie cell segregation.
]]></description>
<dc:creator>Ashlin, T. G.</dc:creator>
<dc:creator>Wu, Z.</dc:creator>
<dc:creator>Xu, Q.</dc:creator>
<dc:creator>Wilkinson, D.</dc:creator>
<dc:date>2019-12-11</dc:date>
<dc:identifier>doi:10.1101/872846</dc:identifier>
<dc:title><![CDATA[Role of SHIP2 in cell repulsion regulated by Eph receptor and ephrin signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2019.12.21.884601v1?rss=1">
<title>
<![CDATA[
A switch in cilia-mediated Hedgehog signaling controls muscle stem cell quiescence and cell cycle progression 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2019.12.21.884601v1?rss=1"
</link>
<description><![CDATA[
Tissue homeostasis requires a tight control of stem cells to maintain quiescence in normal conditions, and ensure a balance between progenitor cell production and the need to preserve a stem cell pool in repair conditions. Using ex-vivo and in-vivo genetic approaches, we provide evidence that primary cilium-mediated repressive Hedgehog (Hh) signalling is required to maintain skeletal muscle stem cells (MuSCs) in a quiescent state. De-repression and further activation of Hh signalling initiates MuSC entry and progression through the cell cycle, and controls self-renewal to ensure efficient repair of injured muscles. We propose a model whereby disassembly of primary cilia upon MuSC activation induces a switch in Hh signalling from a repressive to active state that controls exit from quiescence. Positive Hh response in bi-potential muscle progenitor cells regulates also cell cycle progression and drives MuSC self-renewal. These findings identify Hh signalling as a major regulator of MuSC activity.

HighlightsO_LICilia-containing quiescent MuSCs are Hh signalling suppressed
C_LIO_LIMuSC activation coincides with a switch to active Hh signalling
C_LIO_LISmo mutation delays cell cycle entry and progression, and causes impaired self-renewal
C_LIO_LIPtch1 mutation promotes exit from quiescence, rapid cell cycle and increased self-renewal
C_LI

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=147 HEIGHT=200 SRC="FIGDIR/small/884601v1_ufig1.gif" ALT="Figure 1">
View larger version (31K):
org.highwire.dtl.DTLVardef@1cbaa81org.highwire.dtl.DTLVardef@273898org.highwire.dtl.DTLVardef@f4b26corg.highwire.dtl.DTLVardef@edda2c_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Cruz-Migoni, S. B.</dc:creator>
<dc:creator>Mohd Imran, K.</dc:creator>
<dc:creator>Wahid, A.</dc:creator>
<dc:creator>Rahman, O.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Borycki, A.-G.</dc:creator>
<dc:date>2019-12-22</dc:date>
<dc:identifier>doi:10.1101/2019.12.21.884601</dc:identifier>
<dc:title><![CDATA[A switch in cilia-mediated Hedgehog signaling controls muscle stem cell quiescence and cell cycle progression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2019.12.24.887935v1?rss=1">
<title>
<![CDATA[
Transcription levels of a long noncoding RNA orchestrate opposing regulatory and cell fate outcomes in yeast 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2019.12.24.887935v1?rss=1"
</link>
<description><![CDATA[
Many long noncoding RNAs (lncRNAs) act in cis through transcription-coupled chromatin alterations that drive changes in local gene expression. How some cis-acting lncRNAs promote and others repress gene expression remains poorly understood. Here we report that in S. cerevisiae transcription levels of the lncRNA IRT2, located upstream in the promoter of the inducer of meiosis gene, regulate opposing chromatin and transcription states. Low IRT2 transcription displays enhancer RNA-like features. At these levels, IRT2 promotes histone exchange delivering acetylated histone H3 lysine 56 to chromatin thereby facilitating recruitment of a transcription factor and consequently activating transcription. Conversely, increasing IRT2 transcription enhances chromatin assembly and transcriptional repression. The opposing functions of IRT2 direct a regulatory circuit, which ensures that cells expressing opposite, but not one of either, mating-type loci enter meiosis. Our data demonstrate that the transcription levels of an lncRNA are key to controlling gene expression and cell fate outcomes.
]]></description>
<dc:creator>Moretto, F.</dc:creator>
<dc:creator>Wood, N. E.</dc:creator>
<dc:creator>Chia, M.</dc:creator>
<dc:creator>Li, C.</dc:creator>
<dc:creator>Luscombe, N.</dc:creator>
<dc:creator>van Werven, F.</dc:creator>
<dc:date>2019-12-27</dc:date>
<dc:identifier>doi:10.1101/2019.12.24.887935</dc:identifier>
<dc:title><![CDATA[Transcription levels of a long noncoding RNA orchestrate opposing regulatory and cell fate outcomes in yeast]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2019.12.29.889543v1?rss=1">
<title>
<![CDATA[
Species-specific developmental timing is associated with global differences in protein stability in mouse and human 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2019.12.29.889543v1?rss=1"
</link>
<description><![CDATA[
What determines the pace of embryonic development? Although many molecular mechanisms controlling developmental processes are evolutionarily conserved, the speed at which these operate can vary substantially between species. For example, the same genetic programme, comprising sequential changes in transcriptional states, governs the differentiation of motor neurons in mouse and human, but the tempo at which it operates differs between species. Using in vitro directed differentiation of embryonic stem cells to motor neurons, we show that the programme runs twice as fast in mouse as in human. We provide evidence that this is neither due to differences in signalling, nor the genomic sequence of genes or their regulatory elements. Instead, we find an approximately two-fold increase in protein stability and cell cycle duration in human cells compared to mouse. This can account for the slower pace of human development, indicating that global differences in key kinetic parameters play a major role in interspecies differences in developmental tempo.
]]></description>
<dc:creator>Rayon, T.</dc:creator>
<dc:creator>Stamataki, D.</dc:creator>
<dc:creator>Perez-Carrasco, R.</dc:creator>
<dc:creator>Garcia-Perez, L.</dc:creator>
<dc:creator>Barrington, C.</dc:creator>
<dc:creator>Melchionda, M.</dc:creator>
<dc:creator>Exelby, K.</dc:creator>
<dc:creator>Tybulewicz, V.</dc:creator>
<dc:creator>Fisher, E. M. C.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:date>2019-12-31</dc:date>
<dc:identifier>doi:10.1101/2019.12.29.889543</dc:identifier>
<dc:title><![CDATA[Species-specific developmental timing is associated with global differences in protein stability in mouse and human]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.01.06.896241v1?rss=1">
<title>
<![CDATA[
Regulation of mitophagy by the NSL complex underlies genetic risk for Parkinson's disease at Chr16q11.2 and on the MAPT H1 allele. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.01.06.896241v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is a common incurable neurodegenerative disease. The identification of genetic variants via genome-wide association studies (GWAS) has considerably advanced our understanding of the PD genetic risk. Understanding the functional significance of the risk loci is now a critical step towards translating these genetic advances into an enhanced biological understanding of the disease. Impaired mitophagy is a key causative pathway in familial PD, but its relevance to idiopathic PD is unclear. We used a mitophagy screening assay to evaluate the functional significance of risk genes identified through GWAS. We identified two new regulators of PINK1-mitophagy, KAT8 and KANSL1, previously shown to modulate lysine acetylation. We show that KAT8 and KANSL1 modulate PINK1 gene expression and subsequent PINK1-mitophagy. These findings suggest PINK1-mitophagy is a contributing factor to idiopathic PD. KANSL1 is located on chromosome 17q21 where the risk associated gene has long been considered to be MAPT. While our data does not exclude a possible association between the MAPT gene and PD, it provides strong evidence that KANSL1 plays a crucial role in the disease. Finally, these results enrich our understanding of physiological events regulating mitophagy and establish a novel pathway for drug targeting in neurodegeneration.
]]></description>
<dc:creator>Soutar, M.</dc:creator>
<dc:creator>Melandri, D.</dc:creator>
<dc:creator>Annuario, E.</dc:creator>
<dc:creator>Monaghan, A.</dc:creator>
<dc:creator>Welsh, N.</dc:creator>
<dc:creator>D'Sa, K.</dc:creator>
<dc:creator>Guelfi, S.</dc:creator>
<dc:creator>Zhang, D.</dc:creator>
<dc:creator>Pittman, A.</dc:creator>
<dc:creator>Trabzuni, D.</dc:creator>
<dc:creator>Pan, K.</dc:creator>
<dc:creator>Kia, D. A.</dc:creator>
<dc:creator>Bictash, M.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Houlden, H.</dc:creator>
<dc:creator>Cookson, M. R.</dc:creator>
<dc:creator>Wood, N.</dc:creator>
<dc:creator>Singleton, A.</dc:creator>
<dc:creator>Hardy, J.</dc:creator>
<dc:creator>Whiting, P.</dc:creator>
<dc:creator>Blauwendraat, C.</dc:creator>
<dc:creator>Whitworth, A.</dc:creator>
<dc:creator>Manzoni, C.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:creator>Lewis, P.</dc:creator>
<dc:creator>Plun-Favreau, H.</dc:creator>
<dc:date>2020-01-07</dc:date>
<dc:identifier>doi:10.1101/2020.01.06.896241</dc:identifier>
<dc:title><![CDATA[Regulation of mitophagy by the NSL complex underlies genetic risk for Parkinson's disease at Chr16q11.2 and on the MAPT H1 allele.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.01.07.897694v1?rss=1">
<title>
<![CDATA[
Local retinoic acid directs emergence of the extraocular muscle functional unit 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.01.07.897694v1?rss=1"
</link>
<description><![CDATA[
Coordinated development of muscles, tendons, and their attachment sites ensures emergence of functional musculoskeletal units that are adapted to diverse anatomical demands among different species. How these different tissues are patterned and functionally assembled during embryogenesis is poorly understood. Here, we investigated the morphogenesis of extraocular muscles (EOMs), an evolutionary conserved cranial muscle group that is crucial for the coordinated movement of the eyeballs and for visual acuity. By means of lineage analysis, we redefined the cellular origins of periocular connective tissues interacting with the EOMs, which do not arise exclusively from neural crest mesenchyme as previously thought. Using 3D imaging approaches, we established an integrative blueprint for the EOM functional unit. By doing so, we identified a developmental time window where individual EOMs emerge from a unique muscle anlage and establish insertions in the sclera, which sets these muscles apart from classical muscle-to-bone type of insertions. Further, we demonstrate that the eyeballs are a source of diffusible retinoic acid that allow their targeting by the EOMs in a temporal and dose dependent manner. Using genetically modified mice and inhibitor treatments, we find that endogenous local variations in the concentration of retinoids contribute to the establishment of tendon condensations and attachment sites that precede the initiation of muscle patterning. Collectively, our results highlight how global and site-specific programs are deployed for the assembly of muscle functional units with precise definition of muscle shapes and topographical wiring of their tendon attachments.
]]></description>
<dc:creator>Comai, G. E.</dc:creator>
<dc:creator>Tesarova, M.</dc:creator>
<dc:creator>Dupe, V.</dc:creator>
<dc:creator>Rhinn, M.</dc:creator>
<dc:creator>Vallecillo Garcia, P.</dc:creator>
<dc:creator>da Silva, F.</dc:creator>
<dc:creator>Feret, B.</dc:creator>
<dc:creator>Exelby, K.</dc:creator>
<dc:creator>Dolle, P.</dc:creator>
<dc:creator>Carlsson, L.</dc:creator>
<dc:creator>Pryce, B.</dc:creator>
<dc:creator>Spitz, F.</dc:creator>
<dc:creator>Stricker, S.</dc:creator>
<dc:creator>Zikmund, T.</dc:creator>
<dc:creator>Kaiser, J.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Schedl, A.</dc:creator>
<dc:creator>Ghyselinck, N. B.</dc:creator>
<dc:creator>Schweitzer, R.</dc:creator>
<dc:creator>Tajbakhsh, S.</dc:creator>
<dc:date>2020-01-07</dc:date>
<dc:identifier>doi:10.1101/2020.01.07.897694</dc:identifier>
<dc:title><![CDATA[Local retinoic acid directs emergence of the extraocular muscle functional unit]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.01.06.896571v1?rss=1">
<title>
<![CDATA[
Identifying strategies to target the metabolic flexibility of tumours. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.01.06.896571v1?rss=1"
</link>
<description><![CDATA[
Plasticity of cancer metabolism can be a major obstacle for efficient targeting of tumour-specific metabolic vulnerabilities. Here, we identify and quantify the compensatory mechanisms following the inhibition of major pathways of central carbon metabolism in c-MYC-induced liver tumours. We find that glutaminase isoform Gls2, expressed in normal liver, compensates for the deletion of Gls1 isoform expressed in tumours. Inhibiting both glutaminases significantly delays tumourigenesis but does not completely block glutamine catabolism through the Krebs cycle. We reveal that glutamine catabolism is then driven by amidotransferases. Consistently, the synergistic effect of glutaminase and amidotransferase inhibitors on proliferation of mouse and human tumour cells is observed in vitro and in vivo. Furthermore, when Gls1 is deleted the Krebs cycle activity and tumour formation can also be significantly affected if glycolysis is co-inhibited (Gls1KO/Hk2KO). Finally, the inhibition of either serine (Psat1KO) or fatty acid (FasnKO) biosynthesis can be compensated by uptake of circulating nutrients. Thus, removing these nutrients from the diet produces synergistic effects on suppression of tumourigenesis. These results highlight the high flexibility of tumour metabolism and demonstrate how targeting compensatory mechanisms can improve a therapeutic outcome.
]]></description>
<dc:creator>Yuneva, M.</dc:creator>
<dc:creator>Mendez-Lucas, A.</dc:creator>
<dc:creator>Lin, W.</dc:creator>
<dc:creator>Driscoll, P. C.</dc:creator>
<dc:creator>Legrave, N.</dc:creator>
<dc:creator>Novellasdemunt Vilaseca, L.</dc:creator>
<dc:creator>Xie, C.</dc:creator>
<dc:creator>Charles, M.</dc:creator>
<dc:creator>Wilson, Z.</dc:creator>
<dc:creator>Jones, N. P.</dc:creator>
<dc:creator>Rayport, S. G.</dc:creator>
<dc:creator>Rodriguez-Justo, M.</dc:creator>
<dc:creator>Li, V. S. W.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Hay, N.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:date>2020-01-07</dc:date>
<dc:identifier>doi:10.1101/2020.01.06.896571</dc:identifier>
<dc:title><![CDATA[Identifying strategies to target the metabolic flexibility of tumours.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.01.08.898478v1?rss=1">
<title>
<![CDATA[
Intergenic RNA mainly derives from nascent transcripts of known genes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.01.08.898478v1?rss=1"
</link>
<description><![CDATA[
BackgroundEukaryotic genomes undergo pervasive transcription, leading to the production of many types of stable and unstable RNAs. Transcription is not restricted to regions with annotated gene features but includes almost any genomic context. Currently, the source and function of most RNAs originating from intergenic regions in the human genome remains unclear.

ResultsWe hypothesised that many intergenic RNA can be ascribed to the presence of as-yet unannotated genes or the  fuzzy transcription of known genes that extends beyond the annotated boundaries. To elucidate the contributions of these two sources, we assembled a dataset of >2.5 billion publicly available RNA-seq reads across 5 human cell lines and multiple cellular compartments to annotate transcriptional units in the human genome. About 80% of transcripts from unannotated intergenic regions can be attributed to the fuzzy transcription of existing genes; the remaining transcripts originate mainly from putative long non-coding RNA loci that are rarely spliced. We validated the transcriptional activity of these intergenic RNA using independent measurements, including transcriptional start sites, chromatin signatures, and genomic occupancies of RNA polymerase II in various phosphorylation states. We also analysed the nuclear localisation and sensitivities of intergenic transcripts to nucleases to illustrate that they tend to be rapidly degraded either  on-chromatin by XRN2 or  off-chromatin by the exosome.

ConclusionsWe provide a curated atlas of intergenic RNAs that distinguishes between alternative processing of well annotated genes from independent transcriptional units based on the combined analysis of chromatin signatures, nuclear RNA localisation and degradation pathways.
]]></description>
<dc:creator>Agostini, F.</dc:creator>
<dc:creator>Zagalak, J. A.</dc:creator>
<dc:creator>Attig, J.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:date>2020-01-09</dc:date>
<dc:identifier>doi:10.1101/2020.01.08.898478</dc:identifier>
<dc:title><![CDATA[Intergenic RNA mainly derives from nascent transcripts of known genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.01.10.898726v1?rss=1">
<title>
<![CDATA[
ARID2 deficiency promotes tumor progression and is associated with higher sensitivity to PARP inhibition in lung cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.01.10.898726v1?rss=1"
</link>
<description><![CDATA[
The survival rate in lung cancer remains stubbornly low and there is an urgent need for the identification of new therapeutic targets. Last decades research has evidenced a clear role of chromatin structure in cancer development and several members of the SWI/SNF chromatin remodeling complexes have been described altered in different tumor types. Nevertheless, the precise mechanisms of their impact on cancer progression, as well as the application of this knowledge to cancer patient management are largely unknown.

In this study, we have performed targeted sequencing of a cohort of lung cancer patients on genes involved in chromatin structure, as well as functional experiments to identify the molecular mechanisms linking alterations of chromatin remodeling genes and tumor development.

We have identified ARID2 production loss in 20% of lung cancer patients. Additionally, we have shown that ARID2-deficiency provokes profound chromatin structural changes, alters the transcriptional programme and impairs DNA repair which bolster the proliferative and metastatic potential of the cells both in vitro and in vivo. Moreover, we have demonstrated that ARID2 deficiency significantly affects the sensitivity of the cells to PARP inhibition.

All these results support that ARID2 is a bona-fide tumor suppressor gene in lung cancer that might be exploited therapeutically.
]]></description>
<dc:creator>Moreno, T.</dc:creator>
<dc:creator>Gonzalez-Silva, L.</dc:creator>
<dc:creator>Monterde, B.</dc:creator>
<dc:creator>Betancor-Fernandez, I.</dc:creator>
<dc:creator>Revilla, C.</dc:creator>
<dc:creator>Agraz-Doblas, A.</dc:creator>
<dc:creator>Freire, J.</dc:creator>
<dc:creator>Isidro, P.</dc:creator>
<dc:creator>Quevedo, L.</dc:creator>
<dc:creator>Montes-Moreno, S.</dc:creator>
<dc:creator>Cereceda, L.</dc:creator>
<dc:creator>Astudillo, A.</dc:creator>
<dc:creator>Casar, B.</dc:creator>
<dc:creator>Crespo, P.</dc:creator>
<dc:creator>Morales Torres, C.</dc:creator>
<dc:creator>Scaffidi, P.</dc:creator>
<dc:creator>Gomez-Roman, J.</dc:creator>
<dc:creator>Salido, E.</dc:creator>
<dc:creator>Varela, I.</dc:creator>
<dc:date>2020-01-10</dc:date>
<dc:identifier>doi:10.1101/2020.01.10.898726</dc:identifier>
<dc:title><![CDATA[ARID2 deficiency promotes tumor progression and is associated with higher sensitivity to PARP inhibition in lung cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.01.13.904912v1?rss=1">
<title>
<![CDATA[
Regulated repression, and not activation, governs the cell fate promoter controlling yeast meiosis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.01.13.904912v1?rss=1"
</link>
<description><![CDATA[
Intrinsic signals and cues from the external environment drive cell fate decisions. In budding yeast, the decision to enter meiosis is controlled by nutrient and mating-type signals that regulate expression of the master transcription factor for meiotic entry, IME1. How nutrient signals control IME1 expression remains poorly understood. Here we show that IME1 transcription is regulated by multiple sequence-specific transcription factors that mediate association of Tup1-Cyc8 co-repressor to its promoter. We find that at least eight transcription factors bind the IME1 promoter when nutrients are ample. Remarkably, association of these transcription factors is highly regulated by different nutrient cues. Mutant cells lacking three transcription factors (Sok2/Phd1/Yap6) displayed reduced Tup1-Cyc8 association, increased IME1 expression and earlier onset of meiosis. Our data demonstrate that the promoter of a master regulator is primed for rapid activation while repression by multiple transcription factors mediating Tup1-Cyc8 recruitment dictates the fate decision to enter meiosis.
]]></description>
<dc:creator>Tam, J.</dc:creator>
<dc:creator>van Werven, F.</dc:creator>
<dc:date>2020-01-14</dc:date>
<dc:identifier>doi:10.1101/2020.01.13.904912</dc:identifier>
<dc:title><![CDATA[Regulated repression, and not activation, governs the cell fate promoter controlling yeast meiosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.01.20.912311v1?rss=1">
<title>
<![CDATA[
Super-Beacons: open-source probes with spontaneous tunable blinking compatible with live-cell super-resolution microscopy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.01.20.912311v1?rss=1"
</link>
<description><![CDATA[
Localization based super-resolution microscopy relies on the detection of individual molecules cycling between fluorescent and non-fluorescent states. These transitions are commonly regulated by high-intensity illumination, imposing constrains to imaging hardware and producing sample photodamage. Here, we propose single-molecule self-quenching as a mechanism to generate spontaneous photoswitching independent of illumination. To demonstrate this principle, we developed a new class of DNA-based open-source Super-Resolution probes named Super-Beacons, with photoswitching kinetics that can be tuned structurally, thermally and chemically. The potential of these probes for live-cell friendly Super-Resolution Microscopy without high-illumination or toxic imaging buffers is revealed by imaging Interferon Inducible Transmembrane proteins (IFITMs) at sub-100nm resolutions.
]]></description>
<dc:creator>Pereira, P. M.</dc:creator>
<dc:creator>Gustafsson, N.</dc:creator>
<dc:creator>Marsh, M.</dc:creator>
<dc:creator>Mhlanga, M. M.</dc:creator>
<dc:creator>Henriques, R.</dc:creator>
<dc:date>2020-01-20</dc:date>
<dc:identifier>doi:10.1101/2020.01.20.912311</dc:identifier>
<dc:title><![CDATA[Super-Beacons: open-source probes with spontaneous tunable blinking compatible with live-cell super-resolution microscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.01.19.911891v1?rss=1">
<title>
<![CDATA[
Human AKTIP interacts with ESCRT proteins and functions at the midbody in cytokinesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.01.19.911891v1?rss=1"
</link>
<description><![CDATA[
To complete mitosis, the intercellular bridge that links daughter cells needs to be cleaved. This abscission step is carried out by the sequential recruitment of ESCRT proteins at the midbody. We report here that a new factor, named AKTIP, works in association with ESCRTs. We find that AKTIP binds to the ESCRT I subunit VPS28, and show by high resolution microscopy that AKTIP forms a ring in the dark zone of the intercellular bridge. This ring is positioned in between the circular structures formed by ESCRTs type III. Functionally, we observe that the reduction of AKTIP impinges on the recruitment of the ESCRT III member IST1 at the midbody and causes abscission defects. Taken together, these data indicate that AKTIP is a new factor that contributes to the formation of the ESCRT complex at the midbody and is implicated in the performance of the ESCRT machinery during cytokinetic abscission.
]]></description>
<dc:creator>Merigliano, C.</dc:creator>
<dc:creator>Burla, R.</dc:creator>
<dc:creator>La Torre, M.</dc:creator>
<dc:creator>Del Giudice, S.</dc:creator>
<dc:creator>Teo, H. L.</dc:creator>
<dc:creator>Liew, C. W.</dc:creator>
<dc:creator>Goh, W. I.</dc:creator>
<dc:creator>Chojnowski, A.</dc:creator>
<dc:creator>Olmos, Y.</dc:creator>
<dc:creator>Chiolo, I. E.</dc:creator>
<dc:creator>Carlton, J. G.</dc:creator>
<dc:creator>Raimondo, D.</dc:creator>
<dc:creator>Stewart, C. L.</dc:creator>
<dc:creator>Rhodes, D.</dc:creator>
<dc:creator>Wright, G. D.</dc:creator>
<dc:creator>Burke, B.</dc:creator>
<dc:creator>Saggio, I.</dc:creator>
<dc:date>2020-01-20</dc:date>
<dc:identifier>doi:10.1101/2020.01.19.911891</dc:identifier>
<dc:title><![CDATA[Human AKTIP interacts with ESCRT proteins and functions at the midbody in cytokinesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.01.22.915363v1?rss=1">
<title>
<![CDATA[
Pyphe: A python toolbox for assessing microbial growth and cell viability in high-throughput colony screens 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.01.22.915363v1?rss=1"
</link>
<description><![CDATA[
Microbial fitness screens are a key technique in functional genomics. We present an all-in-one solution, pyphe, for automating and improving data analysis pipelines associated with large-scale fitness screens, including image acquisition and quantification, data normalisation, and statistical analysis. Pyphe is versatile and processes fitness data from colony sizes, viability scores from phloxine B staining or colony growth curves, all obtained with inexpensive transilluminating flatbed scanners. We apply pyphe to show that the fitness information contained in late endpoint measurements of colony sizes is similar to maximum growth slopes from time series. We phenotype gene-deletion strains of fission yeast in 59,350 individual fitness assays in 70 conditions, revealing that colony size and viability provide complementary, independent information. Viability scores obtained from quantifying the redness of phloxine-stained colonies accurately reflect the fraction of live cells within colonies. Pyphe is user-friendly, open-source and fully-documented, illustrated by applications to diverse fitness analysis scenarios.
]]></description>
<dc:creator>Kamrad, S.</dc:creator>
<dc:creator>Rodriguez-Lopez, M.</dc:creator>
<dc:creator>Cotobal, C.</dc:creator>
<dc:creator>Correia-Melo, C.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:creator>Bahler, J.</dc:creator>
<dc:date>2020-01-23</dc:date>
<dc:identifier>doi:10.1101/2020.01.22.915363</dc:identifier>
<dc:title><![CDATA[Pyphe: A python toolbox for assessing microbial growth and cell viability in high-throughput colony screens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.02.09.940536v1?rss=1">
<title>
<![CDATA[
Wheat pathogen Zymoseptoria tritici N-myristoyltransferase inhibitors: on-target antifungal activity and an unusual metabolic defense mechanism 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.02.09.940536v1?rss=1"
</link>
<description><![CDATA[
Zymoseptoria tritici is the causative agent of Septoria tritici blotch (STB), which costs billions of dollars annually to major wheat-producing countries in terms of both fungicide use and crop loss. Agricultural pathogenic fungi have acquired resistance to most commercially available fungicide classes, and the rate of discovery and development of new fungicides has stalled, demanding new approaches and insights. Here we investigate a potential mechanism of targeting an important wheat pathogen Z. tritici via inhibition of N-myristoyltransferase (NMT). We characterize Z. tritici NMT biochemically for the first time, profile the in vivo Z. tritici myristoylated proteome and identify and validate the first Z. tritici NMT inhibitors. Proteomic investigation of the downstream effects of NMT inhibition identified an unusual and novel mechanism of defense against chemical toxicity in Z. tritici through the application of comparative bioinformatics to deconvolute function from the previously largely unannotated Z. tritici proteome. Research into novel fungicidal modes-of-action is essential to satisfy an urgent unmet need for novel fungicide targets, and we anticipate that this study will serve as a useful proteomics and bioinformatics resource for researchers studying Z. tritici.
]]></description>
<dc:creator>Fedoryshchak, R. O.</dc:creator>
<dc:creator>Ocasio, C. A.</dc:creator>
<dc:creator>Strutton, B.</dc:creator>
<dc:creator>Mattocks, J.</dc:creator>
<dc:creator>Corran, A. J.</dc:creator>
<dc:creator>Tate, E. W.</dc:creator>
<dc:date>2020-02-10</dc:date>
<dc:identifier>doi:10.1101/2020.02.09.940536</dc:identifier>
<dc:title><![CDATA[Wheat pathogen Zymoseptoria tritici N-myristoyltransferase inhibitors: on-target antifungal activity and an unusual metabolic defense mechanism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.02.14.928432v1?rss=1">
<title>
<![CDATA[
A divergent cyclin/cyclin-dependent kinase complex controls the atypical replication of Plasmodium berghei during gametogony and parasite transmission. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.02.14.928432v1?rss=1"
</link>
<description><![CDATA[
Cell cycle transitions are generally triggered by variation in the activity of cyclin-dependent kinases (CDKs) bound to cyclins. Malaria-causing parasites have a life cycle with unique cell-division cycles, and a repertoire of divergent CDKs and cyclins of poorly understood function and interdependency. We show that Plasmodium berghei CDK-related kinase 5 (CRK5), is a critical regulator of atypical mitosis in the gametogony and is required for mosquito transmission. It phosphorylates canonical CDK motifs of components in the pre-replicative complex and is essential for DNA replication. During a replicative cycle, CRK5 stably interacts with a single Plasmodium-specific cyclin (SOC2), although we obtained no evidence of SOC2 cycling by transcription, translation or degradation. Our results provide evidence that during Plasmodium male gametogony, this divergent cyclin/CDK pair fills the functional space of other eukaryotic cell-cycle kinases controlling DNA replication.
]]></description>
<dc:creator>Balestra, A. C.</dc:creator>
<dc:creator>Zeeshan, M.</dc:creator>
<dc:creator>Rea, E.</dc:creator>
<dc:creator>Pasquarello, C.</dc:creator>
<dc:creator>Klages, N.</dc:creator>
<dc:creator>Mourier, T.</dc:creator>
<dc:creator>Subudhi, A. K.</dc:creator>
<dc:creator>Arboit, P.</dc:creator>
<dc:creator>Brusini, L. A.</dc:creator>
<dc:creator>Pandey, R.</dc:creator>
<dc:creator>Brady, D.</dc:creator>
<dc:creator>Vaughan, S.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:creator>Pain, A.</dc:creator>
<dc:creator>Ferguson, D.</dc:creator>
<dc:creator>Hainard, A.</dc:creator>
<dc:creator>Tewari, R.</dc:creator>
<dc:creator>Brochet, M.</dc:creator>
<dc:date>2020-02-14</dc:date>
<dc:identifier>doi:10.1101/2020.02.14.928432</dc:identifier>
<dc:title><![CDATA[A divergent cyclin/cyclin-dependent kinase complex controls the atypical replication of Plasmodium berghei during gametogony and parasite transmission.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.02.13.947424v1?rss=1">
<title>
<![CDATA[
Defective heart chamber growth and myofibrillogenesis after knockout of adprhl1 gene function by targeted disruption of the ancestral catalytic active site. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.02.13.947424v1?rss=1"
</link>
<description><![CDATA[
ADP-ribosylhydrolase-like 1 (Adprhl1) is a pseudoenzyme expressed in the developing heart myocardium of all vertebrates. In the amphibian Xenopus laevis, knockdown of the two cardiac Adprhl1 protein species (40 and 23 kDa) causes failure of chamber outgrowth but this has only been demonstrated using antisense morpholinos that interfere with RNA-splicing. Transgenic production of 40 kDa Adprhl1 provides only part rescue of these defects. CRISPR/Cas9 technology now enables targeted mutation of the adprhl1 gene in G0-generation embryos with routine cleavage of all alleles. Testing multiple gRNAs distributed across the locus reveals exonic locations that encode critical amino acids for Adprhl1 function. The gRNA recording the highest frequency of a specific ventricle outgrowth phenotype directs Cas9 cleavage of an exon 6 sequence, where microhomology mediated end-joining biases subsequent DNA repairs towards three small in-frame deletions. Mutant alleles encode discrete loss of 1, 3 or 4 amino acids from a di-arginine (Arg271-Arg272) containing peptide loop at the centre of the ancestral ADP-ribosylhydrolase site. Thus despite lacking catalytic activity, it is the modified (adenosine-ribose) substrate binding cleft of Adprhl1 that fulfils an essential role during heart formation. Mutation results in striking loss of myofibril assembly in ventricle cardiomyocytes. The defects suggest Adprhl1 participation from the earliest stage of cardiac myofibrillogenesis and are consistent with previous MO results and Adprhl1 protein localization to actin filament Z-disc boundaries. A single nucleotide change to the gRNA sequence renders it inactive. Mice lacking Adprhl1 exons 3-4 are normal but production of the smaller ADPRHL1 species is unaffected, providing further evidence that cardiac activity is concentrated at the C-terminal protein portion.

HighlightsComparison of adprhl1 morpholinos. Knockdown of the two Xenopus cardiac Adprhl1 protein species (40 and 23 kDa) causes failure of ventricle outgrowth.

CRISPR/Cas9 targeted gene mutation of adprhl1 with multiple gRNAs reveals exonic locations that encode critical amino acids for Adprhl1 function.

Repair of DSBs at exon 6 yields small in-frame deletions that cause specific ventricle myofibril assembly defects.

The deletions disturb a conserved di-arginine containing peptide loop at the centre of the ancestral substrate binding cleft/ADP-ribosylhydrolase site of this pseudoenzyme.

Mice lacking Adprhl1 exons 3-4 are normal but production of the smaller ADPRHL1 species is unaffected, providing further evidence that cardiac activity is concentrated at the C-terminal protein portion.
]]></description>
<dc:creator>Smith, S. J.</dc:creator>
<dc:creator>Towers, N.</dc:creator>
<dc:creator>Demetriou, K.</dc:creator>
<dc:creator>Mohun, T. J.</dc:creator>
<dc:date>2020-02-14</dc:date>
<dc:identifier>doi:10.1101/2020.02.13.947424</dc:identifier>
<dc:title><![CDATA[Defective heart chamber growth and myofibrillogenesis after knockout of adprhl1 gene function by targeted disruption of the ancestral catalytic active site.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.02.14.949859v1?rss=1">
<title>
<![CDATA[
Enteric glia as a source of neural progenitors in adult zebrafish 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.02.14.949859v1?rss=1"
</link>
<description><![CDATA[
The presence and identity of neural progenitors in the enteric nervous system (ENS) of vertebrates is a matter of intense debate. Here we demonstrate that the non-neuronal ENS cell compartment of teleosts shares molecular and morphological characteristics with mammalian enteric glia but cannot be identified by the expression of canonical glia markers. However, unlike their mammalian counterparts, which are generally quiescent and do not undergo neuronal differentiation during homeostasis, we show that a relatively high proportion of zebrafish enteric glia proliferate under physiological conditions giving rise to progeny that differentiate into enteric neurons. We also provide evidence that, similar to brain neural stem cells, the activation and neuronal differentiation of enteric glia are regulated by Notch signalling. Our experiments reveal remarkable similarities between enteric glia and brain neural stem cells in teleosts and open new possibilities for use of mammalian enteric glia as a potential source of neurons to restore the activity of intestinal neural circuits compromised by injury or disease.
]]></description>
<dc:creator>McCallum, S.</dc:creator>
<dc:creator>Obata, Y.</dc:creator>
<dc:creator>Fourli, E.</dc:creator>
<dc:creator>Boeing, S.</dc:creator>
<dc:creator>Peddie, C. J.</dc:creator>
<dc:creator>Xu, Q.</dc:creator>
<dc:creator>Horswell, S.</dc:creator>
<dc:creator>Kelsh, R.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:creator>Wilkinson, D.</dc:creator>
<dc:creator>Pin, C.</dc:creator>
<dc:creator>Pachnis, V.</dc:creator>
<dc:creator>Heanue, T.</dc:creator>
<dc:date>2020-02-15</dc:date>
<dc:identifier>doi:10.1101/2020.02.14.949859</dc:identifier>
<dc:title><![CDATA[Enteric glia as a source of neural progenitors in adult zebrafish]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.02.18.954289v1?rss=1">
<title>
<![CDATA[
A lipocalin mediates unidirectional haem biomineralization in malaria parasites 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.02.18.954289v1?rss=1"
</link>
<description><![CDATA[
During blood stage development, malaria parasites are challenged with the detoxification of enormous amounts of haem released during the proteolytic catabolism of erythrocytic haemoglobin. They tackle this problem by sequestering haem into bioinert crystals known as haemozoin. The mechanisms underlying this biomineralization process remain enigmatic. Here, we demonstrate that both rodent and human malaria parasite species secrete and internalize a lipocalin-like protein, PV5, to control haem crystallization. Transcriptional deregulation of PV5 in the rodent parasite Plasmodium berghei results in inordinate elongation of haemozoin crystals, while conditional PV5 inactivation in the human malaria agent Plasmodium falciparum causes excessive multi-directional crystal branching. Although haemoglobin processing remains unaffected, PV5-deficient parasites generate less haemozoin. Electron diffraction analysis indicates that despite the distinct changes in crystal morphology neither the crystalline order nor unit cell of haemozoin are affected by impaired PV5 function. Deregulation of PV5 expression renders P. berghei hypersensitive to the antimalarial drugs artesunate, chloroquine, and atovaquone, resulting in accelerated parasite clearance following drug treatment in vivo. Together, our findings demonstrate the Plasmodium-tailored role of a lipocalin family member in haemozoin formation and underscore the haem biomineralization pathway as an attractive target for therapeutic exploitation.

SIGNIFICANCEDuring blood stage development, the malaria parasite replicates inside erythrocytes of the vertebrate host, where it engulfs and digests most of the available haemoglobin. This results in release of the oxygen-binding prosthetic group haem, which is highly toxic in its unbound form. The parasite crystallizes the haem into an insoluble pigment called haemozoin, a process that is vital for parasite survival and which is exploited in antimalarial therapy. We demonstrate that the parasite uses a protein called PV5 in haemozoin formation and that interfering with PV5 expression can increase the parasites sensitivity to antimalarial drugs during blood infection. An improved understanding of the mechanisms underlying haem sequestration will provide valuable insights for future drug development efforts.
]]></description>
<dc:creator>Matz, J. M.</dc:creator>
<dc:creator>Drepper, B.</dc:creator>
<dc:creator>Blum, T. B.</dc:creator>
<dc:creator>van Genderen, E.</dc:creator>
<dc:creator>Burrell, A.</dc:creator>
<dc:creator>Martin, P.</dc:creator>
<dc:creator>Stach, T.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:creator>Abrahams, J. P.</dc:creator>
<dc:creator>Matuschewski, K.</dc:creator>
<dc:creator>Blackman, M. J.</dc:creator>
<dc:date>2020-02-19</dc:date>
<dc:identifier>doi:10.1101/2020.02.18.954289</dc:identifier>
<dc:title><![CDATA[A lipocalin mediates unidirectional haem biomineralization in malaria parasites]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.01.29.918037v1?rss=1">
<title>
<![CDATA[
Patient-specific Alzheimer-like pathology in trisomy 21 cerebral organoids reveals BACE2 as a gene-dose-sensitive AD-suppressor in human brain 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.01.29.918037v1?rss=1"
</link>
<description><![CDATA[
A population of >6 million people worldwide at high risk of Alzheimers disease (AD) are those with Down Syndrome (DS, caused by trisomy 21 (T21)), 70% of whom develop dementia during lifetime, caused by an extra copy of {beta}-amyloid-(A{beta})-precursor-protein gene. We report AD-like pathology in cerebral organoids grown in vitro from non-invasively sampled strands of hair from 71% of DS donors. The pathology consisted of extracellular diffuse and fibrillar A{beta} deposits, hyperphosphorylated/pathologically conformed Tau, and premature neuronal loss.

Presence/absence of AD-like pathology was donor-specific (reproducible between individual organoids/iPSC lines/experiments). Pathology could be triggered in pathology-negative T21 organoids by CRISPR/Cas9-mediated elimination of the third copy of chromosome-21-gene BACE2, but prevented by combined chemical {beta} and {gamma}-secretase inhibition. We found that T21-organoids secrete increased proportions of A{beta}-preventing (A{beta}1-19) and A{beta}-degradation products (A{beta}1-20 and A{beta}1-34). We show these profiles mirror in cerebrospinal fluid of people with DS. We demonstrate that this protective mechanism is mediated by BACE2-trisomy and cross-inhibited by clinically trialled BACE1-inhibitors. Combined, our data prove the physiological role of BACE2 as a dose-sensitive AD-suppressor gene, potentially explaining the dementia delay in [~]30% of people with DS. We also show that DS cerebral organoids could be explored as pre-morbid AD-risk population detector and a system for hypothesis-free drug screens as well as identification of natural suppressor genes for neurodegenerative diseases.
]]></description>
<dc:creator>Alic, I.</dc:creator>
<dc:creator>Goh, P. A.</dc:creator>
<dc:creator>Murray, A.</dc:creator>
<dc:creator>Portelius, E.</dc:creator>
<dc:creator>Gkanatsiou, E.</dc:creator>
<dc:creator>Gough, G.</dc:creator>
<dc:creator>Mok, K. Y.</dc:creator>
<dc:creator>Koschut, D.</dc:creator>
<dc:creator>Brunmeir, R.</dc:creator>
<dc:creator>Yeap, Y. J.</dc:creator>
<dc:creator>O'Brien, N. L.</dc:creator>
<dc:creator>Groet, J.</dc:creator>
<dc:creator>Shao, X.</dc:creator>
<dc:creator>Havlicek, S.</dc:creator>
<dc:creator>Dunn, N. R.</dc:creator>
<dc:creator>Kvartsberg, H.</dc:creator>
<dc:creator>Brinkmalm, G.</dc:creator>
<dc:creator>Hithersay, R.</dc:creator>
<dc:creator>Startin, C.</dc:creator>
<dc:creator>Hamburg, S.</dc:creator>
<dc:creator>Phillips, M.</dc:creator>
<dc:creator>Pervushin, K.</dc:creator>
<dc:creator>Turmaine, M.</dc:creator>
<dc:creator>Wallon, D.</dc:creator>
<dc:creator>Rovelet-Lecrux, A.</dc:creator>
<dc:creator>Soininen, H.</dc:creator>
<dc:creator>Volpi, E.</dc:creator>
<dc:creator>Martin, J. E.</dc:creator>
<dc:creator>Foo, J. N.</dc:creator>
<dc:creator>Becker, D. L.</dc:creator>
<dc:creator>Rostagno, A.</dc:creator>
<dc:creator>Ghiso, J.</dc:creator>
<dc:creator>Krsnik, Z.</dc:creator>
<dc:creator>Simic, G.</dc:creator>
<dc:creator>Kostovic, I.</dc:creator>
<dc:creator>Mitrecic, D.</dc:creator>
<dc:creator>LonDownS Consortium,</dc:creator>
<dc:creator>Francis, P. T.</dc:creator>
<dc:creator>Blennow, K.</dc:creator>
<dc:creator>Strydom, A.</dc:creator>
<dc:creator>Hardy, J.</dc:creator>
<dc:creator>Zetterberg, H.</dc:creator>
<dc:creator>Nizetic, D.</dc:creator>
<dc:date>2020-01-31</dc:date>
<dc:identifier>doi:10.1101/2020.01.29.918037</dc:identifier>
<dc:title><![CDATA[Patient-specific Alzheimer-like pathology in trisomy 21 cerebral organoids reveals BACE2 as a gene-dose-sensitive AD-suppressor in human brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.03.04.972604v1?rss=1">
<title>
<![CDATA[
Visual Biochemistry: modular microfluidics enables kinetic insight from time-resolved cryo-EM 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.03.04.972604v1?rss=1"
</link>
<description><![CDATA[
Mechanistic understanding of biochemical reactions requires structural and kinetic characterization of the underlying chemical processes. However, no single experimental technique can provide this information in a broadly applicable manner and thus structural studies of static macromolecules are often complemented by biophysical analysis. Moreover, the common strategy of utilizing mutants or crosslinking probes to stabilize otherwise short-lived reaction intermediates is prone to trapping off-pathway artefacts and precludes determining the order of molecular events. To overcome these limitations and allow visualisation of biochemical processes at near-atomic spatial resolution and millisecond time scales, we developed a time-resolved sample preparation method for cryo-electron microscopy (trEM). We integrated a modular microfluidic device, featuring a 3D-mixing unit and a delay line of variable length, with a gas-assisted nozzle and motorised plunge-freeze set-up that enables automated, fast, and blot-free sample vitrification. This sample preparation not only preserves high-resolution structural detail but also substantially improves protein distribution across the vitreous ice. We validated the method by examining the formation of RecA filaments on single-stranded DNA. We could reliably visualise reaction intermediates of early filament growth across three orders of magnitude on sub-second timescales. Quantification of the trEM data allowed us to characterize the kinetics of RecA filament growth. The trEM method reported here is versatile, easy to reproduce and thus readily adaptable to a broad spectrum of fundamental questions in biology.
]]></description>
<dc:creator>Maeots, M.-E.</dc:creator>
<dc:creator>Lee, B.</dc:creator>
<dc:creator>Nans, A.</dc:creator>
<dc:creator>Jeong, S.-G.</dc:creator>
<dc:creator>Esfahani, M. M. N.</dc:creator>
<dc:creator>Smith, D. J.</dc:creator>
<dc:creator>Lee, C.-S.</dc:creator>
<dc:creator>Lee, S. S.</dc:creator>
<dc:creator>Peter, M.</dc:creator>
<dc:creator>Enchev, R. I.</dc:creator>
<dc:date>2020-03-05</dc:date>
<dc:identifier>doi:10.1101/2020.03.04.972604</dc:identifier>
<dc:title><![CDATA[Visual Biochemistry: modular microfluidics enables kinetic insight from time-resolved cryo-EM]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.03.05.978478v1?rss=1">
<title>
<![CDATA[
Semantic Segmentation of HeLa Cells: An Objective Comparison between one Traditional Algorithm and Three Deep-Learning Architectures 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.03.05.978478v1?rss=1"
</link>
<description><![CDATA[
In this work, images of a HeLa cancer cell were semantically segmented with one traditional image-processing algorithm and three deep learning architectures: VGG16, ResNet18 and Inception-ResNet-v2. Three hundred slices, each 2000 x 2000 pixels, of a HeLa Cell were acquired with Serial Block Face Scanning Electron Microscopy. The deep learning architectures were pre-trained with ImageNet and then fine-tuned with transfer learning. The image-processing algorithm followed a pipeline of several traditional steps like edge detection, dilation and morphological operators. The algorithms were compared by measuring pixel-based segmentation accuracy and Jaccard index against a labelled ground truth. The results indicated a superior performance of the traditional algorithm (Accuracy = 99%, Jaccard = 93%) over the deep learning architectures: VGG16 (93%, 90%), ResNet18 (94%, 88%), Inception-ResNet-v2 (94%, 89%).
]]></description>
<dc:creator>Karabag, C.</dc:creator>
<dc:creator>Jones, M. L.</dc:creator>
<dc:creator>Peddie, C. J.</dc:creator>
<dc:creator>Weston, A. E.</dc:creator>
<dc:creator>Collinson, L. M.</dc:creator>
<dc:creator>Reyes-Aldasoro, C. C.</dc:creator>
<dc:date>2020-03-05</dc:date>
<dc:identifier>doi:10.1101/2020.03.05.978478</dc:identifier>
<dc:title><![CDATA[Semantic Segmentation of HeLa Cells: An Objective Comparison between one Traditional Algorithm and Three Deep-Learning Architectures]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.03.09.983163v1?rss=1">
<title>
<![CDATA[
Transcriptomic characterization of tuberculous sputum reveals a host Warburg effect and microbial cholesterol catabolism 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.03.09.983163v1?rss=1"
</link>
<description><![CDATA[
The crucial transmission phase of tuberculosis (TB) relies on infectious sputum yet cannot easily be modeled. We applied one-step RNA-Sequencing to sputum from infectious TB patients to investigate the host and microbial environments underlying transmission of Mycobacterium tuberculosis (Mtb). In such TB sputa, compared to non-TB controls, transcriptional upregulation of inflammatory responses and a metabolic shift towards glycolysis was observed in the host. Amongst all bacterial sequences in the sputum, only less than 1.5% originated from Mtb and its abundance is associated with HIV-1 coinfection status. The transcriptome of sputum Mtb more closely resembled aerobic replication and was characterized by evidence of cholesterol utilization, zinc deprivation and reduced expression of the virulence-associated PhoP regulon. Our study provides a comprehensive analysis of the transcriptional landscape associated with infectious sputum and demonstrates the feasibility of applying advanced sequencing technology to readily accessible pathological specimens in the study of host-pathogen adaptation.
]]></description>
<dc:creator>Lai, R. P.</dc:creator>
<dc:creator>Cortes, T.</dc:creator>
<dc:creator>Marais, S.</dc:creator>
<dc:creator>Rockwood, N.</dc:creator>
<dc:creator>Burke, M. L.</dc:creator>
<dc:creator>Garza-Garcia, A.</dc:creator>
<dc:creator>Horswell, S.</dc:creator>
<dc:creator>O'Garra, A.</dc:creator>
<dc:creator>Young, D. B.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:date>2020-03-09</dc:date>
<dc:identifier>doi:10.1101/2020.03.09.983163</dc:identifier>
<dc:title><![CDATA[Transcriptomic characterization of tuberculous sputum reveals a host Warburg effect and microbial cholesterol catabolism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.03.09.967257v1?rss=1">
<title>
<![CDATA[
Identification of Conserved Evolutionary Trajectories in Tumors 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.03.09.967257v1?rss=1"
</link>
<description><![CDATA[
MotivationAs multi-region, time-series, and single cell sequencing data become more widely available, it is becoming clear that certain tumors share evolutionary characteristics with others. In the last few years, several computational methods have been developed with the goal of inferring the subclonal composition and evolutionary history of tumors from tumor biopsy sequencing data. However, the phylogenetic trees that they report differ significantly between tumors (even those with similar characteristics).

ResultsIn this paper, we present a novel combinatorial optimization method, CONETT, for detection of recurrent tumor evolution trajectories. Our method constructs a consensus tree of conserved evolutionary trajectories based on the information about temporal order of alteration events in a set of tumors. We apply our method to previously published datasets of 100 clear-cell renal cell carcinoma and 99 non-small-cell lung cancer patients and identify both conserved trajectories that were reported in the original studies, as well as new trajectories.

AvailabilityCONETT is implemented in C++ and available at https://github.com/ehodzic/CONETT.
]]></description>
<dc:creator>Hodzic, E.</dc:creator>
<dc:creator>Shrestha, R.</dc:creator>
<dc:creator>Malikic, S.</dc:creator>
<dc:creator>Collins, C. C.</dc:creator>
<dc:creator>Litchfield, K.</dc:creator>
<dc:creator>Turajlic, S.</dc:creator>
<dc:creator>Sahinalp, C.</dc:creator>
<dc:date>2020-03-09</dc:date>
<dc:identifier>doi:10.1101/2020.03.09.967257</dc:identifier>
<dc:title><![CDATA[Identification of Conserved Evolutionary Trajectories in Tumors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.03.10.986000v1?rss=1">
<title>
<![CDATA[
LED Zappelin': An open source LED controller for arbitrary spectrum visual stimulation and optogenetics during 2-photon imaging. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.03.10.986000v1?rss=1"
</link>
<description><![CDATA[
Two-photon (2P) microscopy is a cornerstone technique in neuroscience research. However, combining 2P imaging with spectrally arbitrary light stimulation can be challenging due to crosstalk between stimulation light and fluorescence detection. To overcome this limitation, we present a simple and low-cost electronic solution based on an ESP32 microcontroller and a TLC5947 LED driver to rapidly time-interleave stimulation and detection epochs during scans. Implemented for less than $100, our design can independently drive up to 24 arbitrary spectrum LEDs to meet user requirements. We demonstrate the utility of our stimulator for colour vision experiments on the in vivo tetrachromatic zebrafish retina and for optogenetic circuit mapping in Drosophila.

https://github.com/BadenLab/LED-Zappelin
]]></description>
<dc:creator>Zimmermann, M. J.</dc:creator>
<dc:creator>Chagas, A. M.</dc:creator>
<dc:creator>Bartel, P.</dc:creator>
<dc:creator>Pop, S.</dc:creator>
<dc:creator>Prieto-Godino, L. L.</dc:creator>
<dc:creator>Baden, T.</dc:creator>
<dc:date>2020-03-11</dc:date>
<dc:identifier>doi:10.1101/2020.03.10.986000</dc:identifier>
<dc:title><![CDATA[LED Zappelin': An open source LED controller for arbitrary spectrum visual stimulation and optogenetics during 2-photon imaging.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.03.12.987107v1?rss=1">
<title>
<![CDATA[
Progressive changes in hippocampal stem cell properties ensure lifelong neurogenesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.03.12.987107v1?rss=1"
</link>
<description><![CDATA[
Neural stem cell numbers fall rapidly in the hippocampus of juvenile mice but stabilise during adulthood, ensuring lifelong hippocampal neurogenesis. We show that this reduction in stem cell depletion rate is the result of multiple coordinated changes in stem cell behaviour. In particular, while active neural stem cells divide only once or twice before differentiating rapidly in juveniles, they increasingly return to a resting state of shallow quiescence and progress through additional self-renewing divisions in adulthood. Single-cell transcriptomic, mathematical modelling and label-retention analyses indicate that resting cells have a higher activation rate and greater contribution to neurogenesis than dormant cells, which have not left quiescence. These progressive changes in stem cell behaviour result from reduced expression of the pro-activation protein ASCL1 due to increased post-translational degradation. These mechanisms help reconcile current contradictory models of hippocampal NSC dynamics and may contribute to the different rates of decline of hippocampal neurogenesis in mammalian species including humans.
]]></description>
<dc:creator>Harris, L.</dc:creator>
<dc:creator>Rigo, P.</dc:creator>
<dc:creator>Stiehl, T.</dc:creator>
<dc:creator>Gaber, Z.</dc:creator>
<dc:creator>Austin, S. H. L.</dc:creator>
<dc:creator>Masdeu, M. d. M.</dc:creator>
<dc:creator>Edwards, A.</dc:creator>
<dc:creator>Urban, N.</dc:creator>
<dc:creator>Marciniak-Czochra, A.</dc:creator>
<dc:creator>Guillemot, F.</dc:creator>
<dc:date>2020-03-12</dc:date>
<dc:identifier>doi:10.1101/2020.03.12.987107</dc:identifier>
<dc:title><![CDATA[Progressive changes in hippocampal stem cell properties ensure lifelong neurogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.03.12.988956v1?rss=1">
<title>
<![CDATA[
A high-coverage Neandertal genome from Chagyrskaya Cave 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.03.12.988956v1?rss=1"
</link>
<description><![CDATA[
We sequenced the genome of a Neandertal from Chagyrskaya Cave in the Altai Mountains, Russia, to 27-fold genomic coverage. We estimate that this individual lived ~80,000 years ago and was more closely related to Neandertals in western Eurasia (1,2) than to Neandertals who lived earlier in Denisova Cave (3), which is located about 100 km away. About 12.9% of the Chagyrskaya genome is spanned by homozygous regions that are between 2.5 and 10 centiMorgans (cM) long. This is consistent with that Siberian Neandertals lived in relatively isolated populations of less than 60 individuals. In contrast, a Neandertal from Europe, a Denisovan from the Altai Mountains and ancient modern humans seem to have lived in populations of larger sizes. The availability of three Neandertal genomes of high quality allows a first view of genetic features that were unique to Neandertals and that are likely to have been at high frequency among them. We find that genes highly expressed in the striatum in the basal ganglia of the brain carry more amino acid-changing substitutions than genes expressed elsewhere in the brain, suggesting that the striatum may have evolved unique functions in Neandertals.
]]></description>
<dc:creator>Mafessoni, F.</dc:creator>
<dc:creator>Grote, S.</dc:creator>
<dc:creator>de Filippo, C.</dc:creator>
<dc:creator>Slon, V.</dc:creator>
<dc:creator>Kolobova, K. A.</dc:creator>
<dc:creator>Viola, B.</dc:creator>
<dc:creator>Markin, S.</dc:creator>
<dc:creator>Chintalapati, M.</dc:creator>
<dc:creator>Peyregne, S.</dc:creator>
<dc:creator>Skov, L.</dc:creator>
<dc:creator>Skoglund, P.</dc:creator>
<dc:creator>Krivoshapkin, A. I.</dc:creator>
<dc:creator>Derevianko, A. P.</dc:creator>
<dc:creator>Meyer, M.</dc:creator>
<dc:creator>Kelso, J.</dc:creator>
<dc:creator>Peter, B.</dc:creator>
<dc:creator>Prüfer, K.</dc:creator>
<dc:creator>Pääbo, S.</dc:creator>
<dc:date>2020-03-13</dc:date>
<dc:identifier>doi:10.1101/2020.03.12.988956</dc:identifier>
<dc:title><![CDATA[A high-coverage Neandertal genome from Chagyrskaya Cave]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.03.15.992701v1?rss=1">
<title>
<![CDATA[
psiCLIP reveals dynamic RNA binding by DEAH-box helicases before and after exon ligation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.03.15.992701v1?rss=1"
</link>
<description><![CDATA[
Eight RNA helicases remodel the spliceosome to effect pre-mRNA splicing but their mechanism of action remains poorly understood. We have developed "purified spliceosome iCLIP" (psiCLIP) to define helicase-RNA contacts in specific spliceosomal states. psiCLIP reveals previously unappreciated dynamics of spliceosomal helicases. The binding profile of the helicase Prp16 is influenced by the distance between the branch-point and 3 splice site, while Prp22 binds diffusely on the intron before exon ligation but switches to more narrow binding downstream of the exon junction after exon ligation. Notably, depletion of the exon-ligation factor Prp18 destabilizes Prp22 binding to the pre-mRNA, demonstrating that psiCLIP can be used to study the relationships between helicases and auxiliary splicing factors. Thus, psiCLIP is sensitive to spliceosome dynamics and complements the insights from structural and imaging studies by providing crucial positional information on helicase-RNA contacts during spliceosomal remodeling.
]]></description>
<dc:creator>Strittmatter, L. M.</dc:creator>
<dc:creator>Capitanchik, C.</dc:creator>
<dc:creator>Newman, A. J.</dc:creator>
<dc:creator>Hallegger, M.</dc:creator>
<dc:creator>Norman, C. M.</dc:creator>
<dc:creator>Fica, S. M.</dc:creator>
<dc:creator>Oubridge, C.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Nagai, K.</dc:creator>
<dc:date>2020-03-15</dc:date>
<dc:identifier>doi:10.1101/2020.03.15.992701</dc:identifier>
<dc:title><![CDATA[psiCLIP reveals dynamic RNA binding by DEAH-box helicases before and after exon ligation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.03.18.996538v1?rss=1">
<title>
<![CDATA[
A Novel Machine Learning Approach Uncovers New and Distinctive Inhibitors for Cyclin-Dependent Kinase 9 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.03.18.996538v1?rss=1"
</link>
<description><![CDATA[
We present a novel combination of generative and predictive machine learning models for discovering unique protein inhibitors. The new method is assessed on its ability to generate unique inhibitors for the cancer associated protein kinase, CDK9. We validate our method by performing biochemical assays, attaining a hit rate of more than 10%, demonstrating the method to be a notable improvement upon a more standard, and somewhat naive approach. Moreover, we imposed the additional challenge of finding inhibitors that are readily synthesized. Importantly, two new inhibitors are found, with one being distinct from reported CDK9 inhibitors. We discuss the results in the context of modern machine learning principles and the desire expressed by the rational drug design community to secure molecules that are structurally different, yet with high binding affinities, to structurally determined protein targets.
]]></description>
<dc:creator>Assmann, M.</dc:creator>
<dc:creator>Bal, M.</dc:creator>
<dc:creator>Craig, M.</dc:creator>
<dc:creator>D'Oyley, J.</dc:creator>
<dc:creator>Phillips, L.</dc:creator>
<dc:creator>Triendl, H.</dc:creator>
<dc:creator>Bates, P. A.</dc:creator>
<dc:creator>Bashir, U.</dc:creator>
<dc:creator>Ruprah, P.</dc:creator>
<dc:creator>Shaker, N.</dc:creator>
<dc:creator>Stojevic, V.</dc:creator>
<dc:date>2020-03-19</dc:date>
<dc:identifier>doi:10.1101/2020.03.18.996538</dc:identifier>
<dc:title><![CDATA[A Novel Machine Learning Approach Uncovers New and Distinctive Inhibitors for Cyclin-Dependent Kinase 9]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.03.20.000133v1?rss=1">
<title>
<![CDATA[
ZeroCostDL4Mic: an open platform to simplify access and use of Deep-Learning in Microscopy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.03.20.000133v1?rss=1"
</link>
<description><![CDATA[
The resources and expertise needed to use Deep Learning (DL) in bioimaging remain significant barriers for most laboratories. We present https://github.com/HenriquesLab/ZeroCostDL4Mic/wiki, a platform simplifying access to DL by exploiting the free, cloud-based computational resources of Google Colab. https://github.com/HenriquesLab/ZeroCostDL4Mic/wiki allows researchers to train, evaluate, and apply key DL networks to perform tasks including segmentation, detection, denoising, restoration, resolution enhancement and image-to-image translation. We demonstrate the application of the platform to study multiple biological processes.
]]></description>
<dc:creator>Chamier, L. v.</dc:creator>
<dc:creator>Jukkala, J.</dc:creator>
<dc:creator>Spahn, C.</dc:creator>
<dc:creator>Lerche, M.</dc:creator>
<dc:creator>Hernandez-perez, S.</dc:creator>
<dc:creator>Mattila, P.</dc:creator>
<dc:creator>Karinou, E.</dc:creator>
<dc:creator>Holden, S.</dc:creator>
<dc:creator>Can Solak, A.</dc:creator>
<dc:creator>Krull, A.</dc:creator>
<dc:creator>Buchholz, T.-O.</dc:creator>
<dc:creator>Jug, F.</dc:creator>
<dc:creator>Royer, L. A.</dc:creator>
<dc:creator>Heilemann, M.</dc:creator>
<dc:creator>Laine, R. F.</dc:creator>
<dc:creator>Jacquemet, G.</dc:creator>
<dc:creator>Henriques, R.</dc:creator>
<dc:date>2020-03-20</dc:date>
<dc:identifier>doi:10.1101/2020.03.20.000133</dc:identifier>
<dc:title><![CDATA[ZeroCostDL4Mic: an open platform to simplify access and use of Deep-Learning in Microscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.04.02.021733v1?rss=1">
<title>
<![CDATA[
Geometric differences between nuclear envelopes of Wild-type and Chlamydia trachomatis-infected HeLa cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.04.02.021733v1?rss=1"
</link>
<description><![CDATA[
In this work, the geometrical characteristics of two different types of cells observed with Electron Microscopy were analysed. The nuclear envelope of Wild-type HeLa cells and Chlamydia trachomatis-infected HeLa cells were automatically segmented and then modelled against a spheroid and converted to a two-dimensional surface. Geometric measurements from this surface and the volumetric nuclear envelope were extracted to compare the two types of cells. The measurements included the nuclear volume, the sphericity of the nucleus, its flatness or spikiness. In total 13 different cells were segmented: 7 Wild-type and 6 Chlamydia trachomatis-infected. The cells were statistically different in the following measurements. Wild-type HeLa cells have greater volumes than that of Chlamydia trachomatis-infected HeLa cells and they are more spherical as Jaccard index suggests. Standard deviation ({sigma}), and range of values for the nuclear envelope, which shows the distance of the highest peaks and deepest valleys from the spheroid, were also extracted from the modelling against a spheroid and these metrics were used to compare two different data sets in order to draw conclusions.
]]></description>
<dc:creator>Karabag, C.</dc:creator>
<dc:creator>Jones, M. L.</dc:creator>
<dc:creator>Peddie, C. J.</dc:creator>
<dc:creator>Weston, A. E.</dc:creator>
<dc:creator>Collinson, L. M.</dc:creator>
<dc:creator>Reyes-Aldasoro, C. C.</dc:creator>
<dc:date>2020-04-02</dc:date>
<dc:identifier>doi:10.1101/2020.04.02.021733</dc:identifier>
<dc:title><![CDATA[Geometric differences between nuclear envelopes of Wild-type and Chlamydia trachomatis-infected HeLa cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.04.09.035279v1?rss=1">
<title>
<![CDATA[
Differential protein phosphorylation affects the localisation of two secreted Toxoplasma proteins and is widespread during stage conversion 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.04.09.035279v1?rss=1"
</link>
<description><![CDATA[
The intracellular parasite Toxoplasma gondii resides within a membrane bound parasitophorous vacuole (PV) and secretes an array of proteins to establish this replicative niche. It has been shown previously that Toxoplasma both secretes kinases and that numerous proteins are phosphorylated after secretion. Here we assess the role of phosphorylation of SFP1 and the related GRA29, two secreted proteins with unknown function. We show that both proteins form stranded structures in the PV that are independent of the previously described intravacuolar network or actin. GRA29 likely acts as a seed for SFP1 strand formation, and these structures can form independently of other Toxoplasma secreted proteins. We show that an unstructured region at the C-terminus of SFP1 and GRA29 is required for the formation of strands and that mimicking phosphorylation of this domain negatively regulates strand development. When tachyzoites convert to chronic stage bradyzoites, both proteins show a dispersed localisation throughout the cyst matrix. Many secreted proteins are reported to dynamically redistribute as the cyst forms and secreted kinases are known to play a role in cyst formation. Using quantitative phosphoproteome and proteome analysis comparing tachyzoite and early bradyzoite stages, we reveal widespread differential phosphorylation of secreted proteins. These data support a model in which secreted kinases and phosphatases are important to dynamically regulate parasite secreted proteins during stage conversion.

IMPORTANCEToxoplasma gondii is a common parasite that infects up to one third of the human population. Initially the parasite grows rapidly, infecting and destroying cells of the host, but subsequently switches to a slow-growing form and establishes chronic infection. In both stages the parasite lives within a membrane bound vacuole within the host cell, but in the chronic stage a durable cyst wall is synthesized that provides protection to the parasite during transmission to a new host. Toxoplasma secretes proteins into the vacuole to build its replicative niche and previous studies identified many of these proteins as phosphorylated. We investigate two secreted proteins and show that phosphorylation plays an important role in their regulation. We also observed widespread phosphorylation of secreted proteins when parasites convert from acute to chronic stages, providing new insight into how the cyst wall may be dynamically regulated.
]]></description>
<dc:creator>Young, J.</dc:creator>
<dc:creator>Broncel, M.</dc:creator>
<dc:creator>Teague, H.</dc:creator>
<dc:creator>Russell, M. R.</dc:creator>
<dc:creator>McGovern, O.</dc:creator>
<dc:creator>Renshaw, M.</dc:creator>
<dc:creator>Frith, D.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Collinson, L. M.</dc:creator>
<dc:creator>Carruthers, V. B.</dc:creator>
<dc:creator>Ewald, S. E.</dc:creator>
<dc:creator>Treeck, M.</dc:creator>
<dc:date>2020-04-10</dc:date>
<dc:identifier>doi:10.1101/2020.04.09.035279</dc:identifier>
<dc:title><![CDATA[Differential protein phosphorylation affects the localisation of two secreted Toxoplasma proteins and is widespread during stage conversion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.04.12.038414v1?rss=1">
<title>
<![CDATA[
Co-activation of NF-κB and MYC renders cancer cells addicted to IL6 for survival and phenotypic stability 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.04.12.038414v1?rss=1"
</link>
<description><![CDATA[
NF-{kappa}B and MYC are found co-deregulated in human B and plasma-cell cancers. In physiology, NF-{kappa}B is necessary for terminal B-to-plasma cell differentiation, whereas MYC repression is required. It is thus unclear if NF-{kappa}B/MYC co-deregulation is developmentally compatible in carcinogenesis and/or impacts cancer cell differentiation state, possibly uncovering unique sensitivities. Using a mouse system to trace cell lineage and oncogene activation we found that NF-{kappa}B/MYC co-deregulation originated cancers with a plasmablast-like phenotype, alike human plasmablastic-lymphoma and was linked to t(8;14)[MYC-IGH] multiple myeloma. Notably, in contrast to NF-{kappa}B or MYC activation alone, co-deregulation rendered cells addicted to IL6 for survival and phenotypic stability. We propose that conflicting oncogene-driven differentiation pressures can be accommodated at a cost in poorly-differentiated cancers.

SignificanceOur studies improve the understanding of cancer pathogenesis by demonstrating that co-deregulation of NF-{kappa}B and MYC synergize in forming a cancer with a poorly-differentiated state. The cancers in the mouse system share features with human Plasmablastic lymphoma that has a dismal prognosis and no standard of care, and with t(8;14)[MYC-IGH] Multiple myeloma, which is in overall resistant to standard therapy. Notably, we found that NF-{kappa}B and MYC co-deregulation uniquely render cells sensitive to IL6 deprivation, providing a road-map for patient selection. Because of the similarity of the cancers arising in the compound mutant mouse model with that of human Plasmablastic lymphoma and t(8;14)[MYC-IGH] Multiple myeloma, this model could serve in preclinical testing to investigate novel therapies for these hard-to-treat diseases.

HighlightsO_LINF-{kappa}B and MYC co-activation originates (pre)plasmablast-like cancer
C_LIO_LINF-{kappa}B/MYC+ renders cancer cells addicted to IL6 for survival and phenotypic stability
C_LIO_LINF-{kappa}B/MYC+ cancers are alike a fraction of human plasmablastic lymphoma
C_LIO_LIt(8;14)[MYC-IGH] multiple myeloma is linked to a NF-{kappa}B/MYC co-activation signature
C_LI
]]></description>
<dc:creator>Barbosa, R.</dc:creator>
<dc:creator>Xu, A. Q.</dc:creator>
<dc:creator>D'Andrea, D.</dc:creator>
<dc:creator>Copley, F.</dc:creator>
<dc:creator>Patel, H.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Clear, A.</dc:creator>
<dc:creator>Calaminici, M.</dc:creator>
<dc:creator>Janz, M.</dc:creator>
<dc:creator>Zhang, B.</dc:creator>
<dc:creator>Schmidt-Supprian, M.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Gribben, J.</dc:creator>
<dc:creator>Tooze, R.</dc:creator>
<dc:creator>Fitzgibbon, J.</dc:creator>
<dc:creator>Franzoso, G.</dc:creator>
<dc:creator>Rajewsky, K.</dc:creator>
<dc:creator>Calado, D. P.</dc:creator>
<dc:date>2020-04-13</dc:date>
<dc:identifier>doi:10.1101/2020.04.12.038414</dc:identifier>
<dc:title><![CDATA[Co-activation of NF-κB and MYC renders cancer cells addicted to IL6 for survival and phenotypic stability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.04.12.038380v1?rss=1">
<title>
<![CDATA[
Jchain-driven cre enables specific genetic manipulation and timestamping of plasma cell in their niche. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.04.12.038380v1?rss=1"
</link>
<description><![CDATA[
Plasma cells (PC)s are essential for protection from infection, and at the origin of incurable cancers. Current studies do not circumvent limitations of removing PCs from their microenvironment and confound formation and maintenance. This is in part due to the lack of tools to perform specific genetic manipulation in vivo. Also, studies of PC population dynamics have mostly relied on the use of nucleotide analog incorporation that does not label quiescent cells, a property of most PCs. Here we characterize in detail a genetic tool (JchaincreERT2) that permits first-ever specific genetic manipulation in PC in vivo, across immunoglobulin isotypes. Using this tool we found that PC numbers remained constant over-time and that PC decay was compensated by the emergence of new cells, supporting an homeostatic turnover of the population. The JchaincreERT2 genetic tool paves the way for in-depth mechanistic understanding of PC biology and pathology in vivo, in their microenvironment.

HighlightsJchain expression occurs in most plasma cells across immunoglobulin isotypes

JchaincreERT2 mediated genetic manipulation is effective only in plasma cells

Genetic timestamping of plasma cells reveals homeostatic regulation
]]></description>
<dc:creator>Xu, A. Q.</dc:creator>
<dc:creator>Barbosa, R.</dc:creator>
<dc:creator>Calado, D. P.</dc:creator>
<dc:date>2020-04-13</dc:date>
<dc:identifier>doi:10.1101/2020.04.12.038380</dc:identifier>
<dc:title><![CDATA[Jchain-driven cre enables specific genetic manipulation and timestamping of plasma cell in their niche.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.04.14.040634v1?rss=1">
<title>
<![CDATA[
A pan-cancer landscape of somatic substitutions in non-unique regions of the human genome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.04.14.040634v1?rss=1"
</link>
<description><![CDATA[
Around 13% of the human genome displays high sequence similarity with at least one other chromosomal position and thereby poses challenges for computational analyses such as detection of somatic events in cancer. We here extract features of sequencing data from across non-unique regions and employ a machine learning pipeline to describe a landscape of somatic substitutions in 2,658 cancers from the PCAWG cohort. We show mutations in non-unique regions are consistent with mutations in unique regions in terms of mutation load and substitution profiles, and can be validated with linked-read sequencing. This uncovers hidden mutations in ~1,700 coding sequences and thousands of regulatory elements, including known cancer genes, immunoglobulins, and highly mutated gene families.
]]></description>
<dc:creator>Tarabichi, M.</dc:creator>
<dc:creator>Demeulemeester, J.</dc:creator>
<dc:creator>Verfaillie, A.</dc:creator>
<dc:creator>Flanagan, A. M.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>Konopka, T.</dc:creator>
<dc:date>2020-04-15</dc:date>
<dc:identifier>doi:10.1101/2020.04.14.040634</dc:identifier>
<dc:title><![CDATA[A pan-cancer landscape of somatic substitutions in non-unique regions of the human genome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.04.15.042291v1?rss=1">
<title>
<![CDATA[
The deubiquitylase USP9X controls ribosomal stalling 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.04.15.042291v1?rss=1"
</link>
<description><![CDATA[
When a ribosome stalls during translation, it runs the risk of collision with a trailing ribosome. Such an encounter leads to the formation of a stable di-ribosome complex, which needs to be resolved by a dedicated machinery. The initial stalling and the subsequent resolution of di-ribosomal complexes requires activity of Makorin and ZNF598 ubiquitin E3 ligases respectively, through ubiquitylation of the eS10 and uS10 sub-units of the ribosome. It is common for the stability of RING E3 ligases to be regulated by an interacting deubiquitylase (DUB), which often opposes auto-ubiquitylation of the E3. Here, we show that the DUB USP9X directly interacts with ZNF598 and regulates its abundance through the control of protein stability in human cells. We have developed a highly specific small molecule inhibitor of USP9X. Proteomics analysis, following inhibitor treatment of HCT116 cells, confirms previous reports linking USP9X with centrosome associated protein stability and reveals loss of ZNF598 and Makorin 2. In the absence of USP9X or following chemical inhibition of its catalytic activity, steady state levels of Makorins and ZNF598 are diminished and the ribosomal quality control pathway is impaired.
]]></description>
<dc:creator>Clancy, A.</dc:creator>
<dc:creator>Heride, C.</dc:creator>
<dc:creator>Pinto-Fernandez, A.</dc:creator>
<dc:creator>Kallinos, A.</dc:creator>
<dc:creator>Kayser-Bricker, K.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Smith, V.</dc:creator>
<dc:creator>Elcocks, H.</dc:creator>
<dc:creator>Davis, S.</dc:creator>
<dc:creator>Fessler, S.</dc:creator>
<dc:creator>McKinnon, C.</dc:creator>
<dc:creator>Katz, M.</dc:creator>
<dc:creator>Hammonds, T.</dc:creator>
<dc:creator>Jones, N. P.</dc:creator>
<dc:creator>O'Connell, J.</dc:creator>
<dc:creator>Follows, B.</dc:creator>
<dc:creator>Mischke, S.</dc:creator>
<dc:creator>Caravella, J. A.</dc:creator>
<dc:creator>Ioannidis, S.</dc:creator>
<dc:creator>Dinsmore, C.</dc:creator>
<dc:creator>Kim, S.</dc:creator>
<dc:creator>Behrens, A.</dc:creator>
<dc:creator>Komander, D.</dc:creator>
<dc:creator>Kessler, B. M.</dc:creator>
<dc:creator>Urbe, S.</dc:creator>
<dc:creator>Clague, M. J.</dc:creator>
<dc:date>2020-04-16</dc:date>
<dc:identifier>doi:10.1101/2020.04.15.042291</dc:identifier>
<dc:title><![CDATA[The deubiquitylase USP9X controls ribosomal stalling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.04.20.050419v1?rss=1">
<title>
<![CDATA[
Endophilin A2 regulates B cell protein trafficking and humoral responses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.04.20.050419v1?rss=1"
</link>
<description><![CDATA[
Antigen-specific B cell responses require endosomal trafficking to regulate antigen uptake and presentation to helper T cells, and to control expression and signaling of immune receptors. However, the molecular composition of B cell endosomal trafficking pathways and their specific roles in B cell responses have not been systematically investigated. Here we report high-throughput identification of genes regulating B cell receptor (BCR)-mediated antigen internalization using genome-wide functional screens. We show that antigen internalization depends both on clathrin-coated pits and on clathrin-independent endocytosis mediated by endophilin A2. Although endophilin A2 is dispensable for presentation of the endocytosed antigen, it is required for metabolic support of germinal center (GC) B cell proliferation, in part through regulation of iron uptake. Consequently, endophilin A2 deficient mice show selective defects in GC B cell responses and production of high-affinity IgG. The requirement for endophilin A2 highlights a unique importance of clathrin-independent intracellular trafficking in GC B cell clonal expansion and antibody responses.

HIGHLIGHTSO_LIGenome-wide CRISPR screens comprehensively identify genes regulating antigen uptake in B cells
C_LIO_LIB cell receptor-mediated antigen internalization is mediated by both epsin1- dependent clathrin-coated pits and a novel fast endophilin A2-mediated endocytosis.
C_LIO_LIEndophilin A2 is required for peripheral B cell development, antigen-specific germinal center responses and high-affinity IgG production.
C_LIO_LIEndophilin A2 is broadly essential for B cell intracellular trafficking pathways providing metabolic support of germinal center B cell proliferation, in part through regulation of iron uptake.
C_LI
]]></description>
<dc:creator>Malinova, D.</dc:creator>
<dc:creator>Wasim, L.</dc:creator>
<dc:creator>Tolar, P.</dc:creator>
<dc:date>2020-04-20</dc:date>
<dc:identifier>doi:10.1101/2020.04.20.050419</dc:identifier>
<dc:title><![CDATA[Endophilin A2 regulates B cell protein trafficking and humoral responses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.04.21.052944v1?rss=1">
<title>
<![CDATA[
A Structure-Based Mechanism for DNA Entry into the Cohesin Ring 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.04.21.052944v1?rss=1"
</link>
<description><![CDATA[
Despite key roles in sister chromatid cohesion and chromosome organization, the mechanism by which cohesin rings are loaded onto DNA is still unknown. Here, we combine biophysical approaches and cryo-EM to visualize a cohesin loading intermediate in which DNA is locked between two gates that lead into the cohesin ring. Building on this structural framework, we design biochemical experiments to establish the order of events during cohesin loading. In an initial step, DNA traverses an N-terminal kleisin gate that is first opened upon ATP binding and then closed as the cohesin loader locks the DNA against a shut ATPase gate. ATP hydrolysis leads to ATPase gate opening to complete DNA entry. Whether DNA loading is successful, or rather results in loop extrusion, might be dictated by a conserved kleisin N-terminal tail that guides the DNA through the kleisin gate. Our results establish the molecular basis for cohesin loading onto DNA.
]]></description>
<dc:creator>Higashi, T. L.</dc:creator>
<dc:creator>Eickhoff, P.</dc:creator>
<dc:creator>Simoes, J. S.</dc:creator>
<dc:creator>Locke, J.</dc:creator>
<dc:creator>Nans, A.</dc:creator>
<dc:creator>Flynn, H. R.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Papageorgiou, G.</dc:creator>
<dc:creator>O'Reilly, N.</dc:creator>
<dc:creator>Chen, Z. A.</dc:creator>
<dc:creator>O'Reilly, F. J.</dc:creator>
<dc:creator>Rappsilber, J.</dc:creator>
<dc:creator>Costa, A.</dc:creator>
<dc:creator>Uhlmann, F.</dc:creator>
<dc:date>2020-04-21</dc:date>
<dc:identifier>doi:10.1101/2020.04.21.052944</dc:identifier>
<dc:title><![CDATA[A Structure-Based Mechanism for DNA Entry into the Cohesin Ring]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.04.22.054791v1?rss=1">
<title>
<![CDATA[
Unifying the mechanism of mitotic exit control in a spatio-temporal logical model 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.04.22.054791v1?rss=1"
</link>
<description><![CDATA[
1The transition from mitosis into the first gap phase of the cell cycle in budding yeast is controlled by the Mitotic Exit Network (MEN). The network interprets spatio-temporal cues about the progression of mitosis and ensures that release of Cdc14 phosphatase occurs only after completion of key mitotic events. The MEN has been studied intensively however a unified understanding of how localization and protein activity function together as a system is lacking. In this paper we present a compartmental, logical model of the MEN that is capable of representing spatial aspects of regulation in parallel to control of enzymatic activity. Through optimization of the model, we reveal insights into role of Cdc5 in Cdc15 localization and the importance of Lte1 regulation in control of Bfa1. We show that our model is capable of correctly predicting the phenotype of[~] 80% of mutants we tested, including mutants representing mislocalizing proteins. We use a continuous time implementation of the model to demonstrate the role of Cdc14 Early Anaphase Release (FEAR) to ensure robust timing of anaphase and verify our findings in living cells. We show that our model can represent measured cell-cell variation in Spindle Position Checkpoint (SPoC) mutants. Finally, we use the model to predict the impact of forced localization of MEN proteins and validate these predictions experimentally. This model represents a unified view of the mechanism of mitotic exit control.
]]></description>
<dc:creator>Howell, R. S.</dc:creator>
<dc:creator>Klemm, C.</dc:creator>
<dc:creator>Thorpe, P. H.</dc:creator>
<dc:creator>Csikasz-Nagy, A.</dc:creator>
<dc:date>2020-04-23</dc:date>
<dc:identifier>doi:10.1101/2020.04.22.054791</dc:identifier>
<dc:title><![CDATA[Unifying the mechanism of mitotic exit control in a spatio-temporal logical model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.04.23.054833v1?rss=1">
<title>
<![CDATA[
Ribosome profiling reveals ribosome stalling on tryptophan codons upon oxidative stress in fission yeast 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.04.23.054833v1?rss=1"
</link>
<description><![CDATA[
Modulation of translation is an essential response to stress conditions. We have investigated the translational programmes launched by the fission yeast Schizosaccharomyces pombe subject to five environmental stresses: oxidative stress, heavy metal, heat shock, osmotic shock and DNA damage. We also explored the contribution of two major defence pathways to these programmes: The Integrated Stress Response, which directly regulates translation initiation, and the stress-response MAPK pathway. To obtain a genome-wide and high-resolution view of this phenomenon, we performed ribosome profiling of control cells and of cells subject to each of the five stresses mentioned above, both in wild type background and in cells in which the Integrated Stress Response or the MAPK pathway were inactivated.

Translational changes were partially dependent on the integrity of both signalling pathways. Interestingly, we found that the transcription factor Fil1, a functional homologue of the Gcn4 and Atf4 proteins (from budding yeast and mammals, respectively), was highly upregulated in most stresses. Consistent with this result, Fil1 was required for the normal response to most stresses. A large group of mRNAs were translationally downregulated, including many required for ribosome biogenesis. Overall, our data suggest that severe stresses lead to the implementation of a universal translational response, which includes energy-saving measures (reduction of ribosome production) and induction of a Fil1-mediated transcriptional programme.

Surprisingly, ribosomes stalled on tryptophan codons specifically upon oxidative stress, a phenomenon that is likely caused by a decrease in charged tRNA-Tryptophan. Tryptophan stalling led to a mild translation elongation reduction and contributed to the inhibition of initiation by the Integrated Stress Response. Taken together, our results show that different stresses elicit common and specific translational responses, revealing a special and so far unknown role in Tryptophan-tRNA availability.
]]></description>
<dc:creator>Rubio, A.</dc:creator>
<dc:creator>Ghosh, S.</dc:creator>
<dc:creator>Mulleder, M.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:creator>Mata, J.</dc:creator>
<dc:date>2020-04-24</dc:date>
<dc:identifier>doi:10.1101/2020.04.23.054833</dc:identifier>
<dc:title><![CDATA[Ribosome profiling reveals ribosome stalling on tryptophan codons upon oxidative stress in fission yeast]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.04.23.057208v1?rss=1">
<title>
<![CDATA[
Metabolic precision labeling enables selective probing of O-linked N-acetylgalactosamine glycosylation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.04.23.057208v1?rss=1"
</link>
<description><![CDATA[
Protein glycosylation events that happen early in the secretory pathway are often dysregulated during tumorigenesis. These events can be probed, in principle, by monosaccharides with bioorthogonal tags that would ideally be specific for distinct glycan subtypes. However, metabolic interconversion into other monosaccharides drastically reduces such specificity in the living cell. Here, we use a structure-based design process to develop the monosaccharide probe GalNAzMe that is specific for cancer-relevant Ser/Thr-N-acetylgalactosamine (O-GalNAc) glycosylation. By virtue of a branched N-acylamide side chain, GalNAzMe is not interconverted by epimerization to the corresponding N-acetylglucosamine analog like conventional GalNAc-based probes. GalNAzMe enters O-GalNAc glycosylation but does not enter other major cell surface glycan types including Asn (N)-linked glycans. We equip cells with the capacity to biosynthesize the nucleotide-sugar donor UDP-GalNAzMe from a caged precursor. Tagged with a bioorthogonal azide group, GalNAzMe serves as an O-glycan specific reporter in superresolution microscopy, chemical glycoproteomics, a genome-wide CRISPR knock-out (KO) screen, and imaging of intestinal organoids. GalNAzMe is a precision tool that allows a detailed view into the biology of a major type of cancer-relevant protein glycosylation.

Significance statementA large portion of all secreted and cell surface proteins in humans are modified by Ser/Thr(O)-linked glycosylation with N-acetylgalactosamine (GalNAc). While of fundamental importance in health and disease, O-GalNAc glycosylation is technically challenging to study because of a lack of specific tools to be used in biological assays. Here, we design an O-GalNAc specific reporter molecule termed GalNAzMe to selectively label O-GalNAc glycoproteins in living human cells. GalNAzMe is compatible with a range of experiments in quantitative biology to broaden our understanding of glycosylation. We further demonstrate that labeling is genetically programmable by expression of a mutant glycosyltransferase, allowing application even to experiments with low inherent sensitivity.
]]></description>
<dc:creator>Debets, M. F.</dc:creator>
<dc:creator>Tastan, O. Y.</dc:creator>
<dc:creator>Wisnovsky, S. P.</dc:creator>
<dc:creator>Malaker, S. A.</dc:creator>
<dc:creator>Angelis, N.</dc:creator>
<dc:creator>Moeckl, L. K. R.</dc:creator>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>Flynn, H.</dc:creator>
<dc:creator>Wagner, L. J. S.</dc:creator>
<dc:creator>Bineva-Todd, G.</dc:creator>
<dc:creator>Antononopoulos, A.</dc:creator>
<dc:creator>Cioce, A.</dc:creator>
<dc:creator>Browne, W. M.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Briggs, D. C.</dc:creator>
<dc:creator>Douglas, H. L.</dc:creator>
<dc:creator>Hess, G. T.</dc:creator>
<dc:creator>Agbay, A. J.</dc:creator>
<dc:creator>Roustan, C.</dc:creator>
<dc:creator>Kjaer, S.</dc:creator>
<dc:creator>Haslam, S. M.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Bassik, M. C.</dc:creator>
<dc:creator>Moerner, W. E.</dc:creator>
<dc:creator>Li, V. S. W.</dc:creator>
<dc:creator>Bertozzi, C. R.</dc:creator>
<dc:creator>Schumann, B.</dc:creator>
<dc:date>2020-04-25</dc:date>
<dc:identifier>doi:10.1101/2020.04.23.057208</dc:identifier>
<dc:title><![CDATA[Metabolic precision labeling enables selective probing of O-linked N-acetylgalactosamine glycosylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.04.25.061564v1?rss=1">
<title>
<![CDATA[
Ultra-sparse connectivity within the lateral hypothalamus 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.04.25.061564v1?rss=1"
</link>
<description><![CDATA[
The lateral hypothalamus (LH) contains neuronal populations which generate fundamental behavioural actions such as feeding, sleep, movement, attack and evasion. Their activity is also correlated with various appetitive and consummatory behaviours as well as reward seeking. It is unknown how neural activity within and among these populations is coordinated. One hypothesis postulates that they communicate using inhibitory and excitatory synapses, forming local microcircuits. We inspected this hypothesis using quadruple whole cell recordings and optogenetics to screen thousands of potential connections in brain slices. In contrast to the neocortex, we found near zero connectivity within the LH. In line with its ultra-sparse intrinsic connectivity, we found that the LH does not generate local beta and gamma oscillations. This suggests that LH neurons integrate incoming input within individual neurons rather than through local network interactions, and that input from other brain structures is decisive for selecting active populations in LH.
]]></description>
<dc:creator>Burdakov, D.</dc:creator>
<dc:creator>Karnani, M. M.</dc:creator>
<dc:date>2020-04-27</dc:date>
<dc:identifier>doi:10.1101/2020.04.25.061564</dc:identifier>
<dc:title><![CDATA[Ultra-sparse connectivity within the lateral hypothalamus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.04.28.066043v1?rss=1">
<title>
<![CDATA[
Extensive transcriptional and chromatin changes underlie astrocyte maturation in vivo and in culture 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.04.28.066043v1?rss=1"
</link>
<description><![CDATA[
Astrocytes have diverse functions in brain homeostasis. Many of these functions are acquired during late stages of differentiation when astrocytes become fully mature. The mechanisms underlying astrocyte maturation are not well understood. Here we identified extensive transcriptional changes that occur during astrocyte maturation and are accompanied by chromatin remodelling at enhancer elements. Investigating astrocyte maturation in a cell culture model revealed that in vitro-differentiated astrocytes lacked expression of many mature astrocyte-specific genes, including genes for the transcription factors Rorb, Dbx2, Lhx2 and Fezf2. Forced expression of these factors in vitro induced distinct sets of mature astrocytes-specific transcripts. Culturing astrocytes with FGF2 in a three-dimensional gel induced expression of Rorb, Dbx2 and Lhx2 and improved their maturity based on transcriptional and chromatin profiles. Therefore extrinsic signals orchestrate the expression of multiple intrinsic regulators, which in turn induce in a modular manner the transcriptional and chromatin changes underlying astrocyte maturation.
]]></description>
<dc:creator>Lattke, M.</dc:creator>
<dc:creator>Goldstone, R.</dc:creator>
<dc:creator>Guillemot, F.</dc:creator>
<dc:date>2020-04-30</dc:date>
<dc:identifier>doi:10.1101/2020.04.28.066043</dc:identifier>
<dc:title><![CDATA[Extensive transcriptional and chromatin changes underlie astrocyte maturation in vivo and in culture]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.05.01.071704v1?rss=1">
<title>
<![CDATA[
Cryo-EM of human Arp2/3 complexes provides structural insights into actin nucleation modulation by ARPC5 isoforms 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.05.01.071704v1?rss=1"
</link>
<description><![CDATA[
The Arp2/3 complex regulates many cellular processes by stimulating formation of branched actin filament networks. Because three of its seven subunits exist as two different isoforms, mammals produce a family of Arp2/3 complexes with different properties that may be suited to different physiological contexts. To shed light on how isoform diversification affects Arp2/3 function, we determined a 4.2 [A] resolution cryo-EM structure of the most active human Arp2/3 complex containing ARPC1B and ARPC5L, and compared it with the structure of the least active ARPC1A-ARPC5-containing complex. The architecture of each isoform-specified Arp2/3 is the same. Strikingly, however, the N-terminal half of ARPC5L is partially disordered compared to ARPC5, suggesting that this region of ARPC5/ARPC5L is an important determinant of complex activity. Confirming this idea, the nucleation activity of Arp2/3 complexes containing hybrid ARPC5/ARPC5L subunits is higher when the ARPC5L N-terminus is present, thereby explaining activity differences between the different Arp2/3 complexes.
]]></description>
<dc:creator>von Loeffelholz, O.</dc:creator>
<dc:creator>Purkiss, A.</dc:creator>
<dc:creator>Cao, L.</dc:creator>
<dc:creator>Kjaer, S.</dc:creator>
<dc:creator>Kogata, N.</dc:creator>
<dc:creator>Romet-Lemonne, G.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:creator>Moores, C. A.</dc:creator>
<dc:date>2020-05-01</dc:date>
<dc:identifier>doi:10.1101/2020.05.01.071704</dc:identifier>
<dc:title><![CDATA[Cryo-EM of human Arp2/3 complexes provides structural insights into actin nucleation modulation by ARPC5 isoforms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.05.01.072637v1?rss=1">
<title>
<![CDATA[
Gliding motility of Plasmodium merozoites 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.05.01.072637v1?rss=1"
</link>
<description><![CDATA[
Plasmodium malaria parasites are obligate intracellular protozoans that use a unique form of locomotion, termed gliding motility, to move through host tissues and invade cells. The process is substrate-dependent and powered by an actomyosin motor that drives the posterior translocation of extracellular adhesins which in turn propel the parasite forward. Gliding motility is essential for tissue translocation in the sporozoite and ookinete stages; however, the short-lived erythrocyte-invading merozoite stage has never been observed to undergo gliding movement. Here we show Plasmodium merozoites possess the ability to undergo gliding motility and that this mechanism is likely an important precursor step for successful parasite invasion. We demonstrate that two human infective species, P. falciparum and P. knowlesi, have distinct merozoite motility profiles which may reflect distinct invasion strategies. Additionally, we develop and validate a higher throughput assay to evaluate the effects of genetic and pharmacological perturbations on both the molecular motor and complex signaling cascade that regulates motility in merozoites. The discovery of merozoite motility provides a new model to study the glideosome and may facilitate the pursuit of new targets for malaria treatment.

Significance statementPlasmodium malaria parasites use a unique substrate-dependent locomotion termed gliding motility to translocate through tissues and invade cells. Dogma has suggested that the small labile invasive stages that invade erythrocytes, merozoites, use this motility solely to penetrate target erythrocytes. Here we reveal that merozoites use gliding motility for translocation across host cells prior to invasion. This forms an important pre-invasion step that is powered by a conserved actomyosin motor and is regulated by a complex signaling pathway. This work fundamentally changes our understanding of the role of gliding motility and invasion in the blood and will have a significant impact on our understanding of blood stage host-pathogen interactions, parasite biology, and could have implications for vaccine development.
]]></description>
<dc:creator>Yahata, K.</dc:creator>
<dc:creator>Hart, M. N.</dc:creator>
<dc:creator>Davies, H.</dc:creator>
<dc:creator>Asada, M.</dc:creator>
<dc:creator>Templeton, T. J.</dc:creator>
<dc:creator>Treeck, M.</dc:creator>
<dc:creator>Moon, R. W.</dc:creator>
<dc:creator>Kaneko, O.</dc:creator>
<dc:date>2020-05-03</dc:date>
<dc:identifier>doi:10.1101/2020.05.01.072637</dc:identifier>
<dc:title><![CDATA[Gliding motility of Plasmodium merozoites]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.05.04.070177v1?rss=1">
<title>
<![CDATA[
RNA binding regulates TRIM25-mediated RIG-I ubiquitylation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.05.04.070177v1?rss=1"
</link>
<description><![CDATA[
TRIM25 is a ubiquitin E3 ligase active in innate immunity and cell fate decisions. Mounting evidence suggests that TRIM25's E3 ligase activity is regulated by RNAs. However, while mutations affecting RNA binding have been described, neither the precise RNA binding site has been identified nor which domains are involved. Here, we present biophysical evidence for the presence of RNA binding sites on both TRIM25 PRY/SPRY and coiled-coil domains, and map the binding site on the PRY/SPRY with residue resolution. Cooperative RNA-binding of both domains enhances their otherwise transient interaction in solution and increases the E3 ligase activity of TRIM25. We also show that TRIM25 not only binds RNA in mammalian cells but that interfering with RNA binding has an effect on cellular RIG-I ubiquitination.
]]></description>
<dc:creator>Haubrich, K.</dc:creator>
<dc:creator>Augsten, S.</dc:creator>
<dc:creator>Simon, B.</dc:creator>
<dc:creator>Masiewicz, P.</dc:creator>
<dc:creator>Perez, K.</dc:creator>
<dc:creator>Lethier, M.</dc:creator>
<dc:creator>Rittinger, K.</dc:creator>
<dc:creator>Gabel, F.</dc:creator>
<dc:creator>Cusack, S.</dc:creator>
<dc:creator>Hennig, J.</dc:creator>
<dc:date>2020-05-05</dc:date>
<dc:identifier>doi:10.1101/2020.05.04.070177</dc:identifier>
<dc:title><![CDATA[RNA binding regulates TRIM25-mediated RIG-I ubiquitylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.05.05.078360v1?rss=1">
<title>
<![CDATA[
Type I and III interferons disrupt lung epithelial repair during recovery from viral infection 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.05.05.078360v1?rss=1"
</link>
<description><![CDATA[
Excessive cytokine signalling frequently exacerbates lung tissue damage during respiratory viral infection. Type I and III interferons (IFN-/{beta} and IFN-{lambda}) are host-produced antiviral cytokines and currently considered as COVID-19 therapy. Prolonged IFN-/{beta} responses can lead to harmful proinflammatory effects, whereas IFN-{lambda} mainly signals in epithelia, inducing localised antiviral immunity. Here we show that IFN signalling interferes with lung repair during influenza recovery, with IFN-{lambda} driving these effects most potently. IFN-induced p53 directly reduces epithelial proliferation and differentiation, increasing disease severity and susceptibility to bacterial superinfections. Hence, excessive or prolonged IFN-production aggravates viral infection by impairing lung epithelial regeneration. Therefore, timing and duration are critical parameters of endogenous IFN action, and should be considered carefully for IFN therapeutic strategies against viral infections like influenza and COVID-19.

One Sentence SummaryA novel IFN-mediated mechanism of immunopathology during respiratory virus infection by interference with lung tissue repair.
]]></description>
<dc:creator>Major, J.</dc:creator>
<dc:creator>Crotta, S.</dc:creator>
<dc:creator>Llorian, M.</dc:creator>
<dc:creator>McCabe, T. M.</dc:creator>
<dc:creator>Gad, H. H.</dc:creator>
<dc:creator>Hartmann, R.</dc:creator>
<dc:creator>Wack, A.</dc:creator>
<dc:date>2020-05-05</dc:date>
<dc:identifier>doi:10.1101/2020.05.05.078360</dc:identifier>
<dc:title><![CDATA[Type I and III interferons disrupt lung epithelial repair during recovery from viral infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.05.07.082032v1?rss=1">
<title>
<![CDATA[
Lack of adaptation to centriolar defects leads to p53-independent microcephaly in the absence of Cep135 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.05.07.082032v1?rss=1"
</link>
<description><![CDATA[
Autosomal Recessive Primary Microcephaly (MCPH) is a rare disease associated to proteins involved in centrosomal and spindle dynamics including Cep135 (MCPH8). Although Cep135 has been associated to centriolar assembly, the mechanisms associated to the pathogenesis underlying MCPH8 mutations are unclear. By using a series of CRISPR/Cas9-edited murine Cep135 alleles, we report here that lack of Cep135 results in perinatal lethality accompanied by significant microcephaly in a dosis-dependent manner. Cep135 deficiency, but not that of other centrosomal microcephaly proteins such as Aspm or Cdk5rap2, induces centrosome duplication defects, and perturbed centriole structure and dynamics. Whereas other cell types are able to quickly adapt to these defects, neural progenitors display a prolonged response leading to chromosomal instability and cell death in later developmental stages. Genetic ablation of Trp53 in these mutant embryos prevents apoptotic cell death but does not rescue the microcephaly induced by Cep135 loss. These results suggest that microcephaly can arise from the lack of adaptation to centriole defects in neural progenitors of the developing neocortex in a p53-independent manner.
]]></description>
<dc:creator>Malumbres, M.</dc:creator>
<dc:creator>Gonzalez-Martinez, J.</dc:creator>
<dc:creator>Cwersch, A.</dc:creator>
<dc:creator>Martinez-Alonso, D.</dc:creator>
<dc:creator>Lopez, L. R.</dc:creator>
<dc:creator>Almagro, J.</dc:creator>
<dc:creator>Megias, D.</dc:creator>
<dc:creator>Boskovic, J.</dc:creator>
<dc:creator>Gilabert, J.</dc:creator>
<dc:creator>Grana Castro, O.</dc:creator>
<dc:creator>Pierani, A.</dc:creator>
<dc:creator>Behrens, A.</dc:creator>
<dc:creator>Ortega, S.</dc:creator>
<dc:date>2020-05-08</dc:date>
<dc:identifier>doi:10.1101/2020.05.07.082032</dc:identifier>
<dc:title><![CDATA[Lack of adaptation to centriolar defects leads to p53-independent microcephaly in the absence of Cep135]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.05.11.088203v1?rss=1">
<title>
<![CDATA[
Genetic determinants of receptor-binding preference and zoonotic potential of H9N2 avian influenza viruses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.05.11.088203v1?rss=1"
</link>
<description><![CDATA[
Receptor recognition and binding is the first step of viral infection and a key determinant of host specificity. The inability of avian influenza viruses to effectively bind human-like sialylated receptors is a major impediment to their efficient transmission in humans and pandemic capacity. Influenza H9N2 viruses are endemic in poultry across Asia and parts of Africa where they occasionally infect humans and are therefore considered viruses with zoonotic potential. We previously described H9N2 viruses, including several isolated from human zoonotic cases, showing a preference for human-like receptors. Here we take a mutagenesis approach, making viruses with single or multiple substitutions in H9 haemagglutinin to determine the genetic basis of preferences for alternative avian receptors and for human-like receptors. We describe amino acid motifs at positions 190, 226 and 227 that play a major role in determining receptor specificity, and several other residues such as 159, 188, 193, 196, 198 and 225 play a smaller role. Furthermore, we show changes at residues 135, 137, 147, 157, 158, 184, 188, and 192 can also modulate virus receptor avidity and that substitutions that increased or decreased the net positive charge around the haemagglutinin receptor-binding site show increases and decreases in avidity, respectively. The motifs we identify as increasing preference for the human-receptor will help guide future H9N2 surveillance efforts and facilitate our understanding of the emergence of influenza viruses with high zoonotic potential.

Author SummaryAs of 2020, over 60 infections of humans by H9N2 influenza viruses have been recorded in countries were the virus is endemic. Avian-like cellular receptors are the primary target for these viruses. However, given that human infections have been detected on an almost monthly basis since 2015, there may be a capacity for H9N2 viruses to evolve and gain the ability to target human-like cellular receptors. Here we identify molecular signatures that can cause viruses to bind human-like receptors, and we identify the molecular basis for the distinctive preference for sulphated receptors displayed by the majority of recent H9N2 viruses. This work will help guide future surveillance by providing markers that signify the emergence of viruses with enhanced zoonotic potential as well as improving understanding the basis of influenza virus receptor-binding.
]]></description>
<dc:creator>Peacock, T. P.</dc:creator>
<dc:creator>Sealy, J. E.</dc:creator>
<dc:creator>Harvey, W. T.</dc:creator>
<dc:creator>Benton, D. J.</dc:creator>
<dc:creator>Reeve, R.</dc:creator>
<dc:creator>Iqbal, M.</dc:creator>
<dc:date>2020-05-12</dc:date>
<dc:identifier>doi:10.1101/2020.05.11.088203</dc:identifier>
<dc:title><![CDATA[Genetic determinants of receptor-binding preference and zoonotic potential of H9N2 avian influenza viruses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.05.14.095414v1?rss=1">
<title>
<![CDATA[
Pre-existing and de novo humoral immunity to SARS-CoV-2 in humans 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.05.14.095414v1?rss=1"
</link>
<description><![CDATA[
Several related human coronaviruses (HCoVs) are endemic in the human population, causing mild respiratory infections1. Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the etiologic agent of Coronavirus disease 2019 (COVID-19), is a recent zoonotic infection that has quickly reached pandemic proportions2,3. Zoonotic introduction of novel coronaviruses is thought to occur in the absence of pre-existing immunity in the target human population. Using diverse assays for detection of antibodies reactive with the SARS-CoV-2 spike (S) glycoprotein, we demonstrate the presence of pre-existing humoral immunity in uninfected and unexposed humans to the new coronavirus. SARS-CoV-2 S-reactive antibodies were readily detectable by a sensitive flow cytometry-based method in SARS-CoV-2-uninfected individuals and were particularly prevalent in children and adolescents. These were predominantly of the IgG class and targeted the S2 subunit. In contrast, SARS-CoV-2 infection induced higher titres of SARS-CoV-2 S-reactive IgG antibodies, targeting both the S1 and S2 subunits, as well as concomitant IgM and IgA antibodies, lasting throughout the observation period of 6 weeks since symptoms onset. SARS-CoV-2-uninfected donor sera also variably reacted with SARS-CoV-2 S and nucleoprotein (N), but not with the S1 subunit or the receptor binding domain (RBD) of S on standard enzyme immunoassays. Notably, SARS-CoV-2-uninfected donor sera exhibited specific neutralising activity against SARS-CoV-2 and SARS-CoV-2 S pseudotypes, according to levels of SARS-CoV-2 S-binding IgG and with efficiencies comparable to those of COVID-19 patient sera. Distinguishing pre-existing and de novo antibody responses to SARS-CoV-2 will be critical for our understanding of susceptibility to and the natural course of SARS-CoV-2 infection.
]]></description>
<dc:creator>Ng, K.</dc:creator>
<dc:creator>Faulkner, N.</dc:creator>
<dc:creator>Cornish, G.</dc:creator>
<dc:creator>Rosa, A.</dc:creator>
<dc:creator>Earl, C.</dc:creator>
<dc:creator>Wrobel, A.</dc:creator>
<dc:creator>Benton, D.</dc:creator>
<dc:creator>Roustan, C.</dc:creator>
<dc:creator>Bolland, W.</dc:creator>
<dc:creator>Thompson, R.</dc:creator>
<dc:creator>Agua-Doce, A.</dc:creator>
<dc:creator>Hobson, P.</dc:creator>
<dc:creator>Heaney, J.</dc:creator>
<dc:creator>Rickman, H.</dc:creator>
<dc:creator>Paraskevopoulou, S.</dc:creator>
<dc:creator>Houlihan, C. F.</dc:creator>
<dc:creator>Thomson, K.</dc:creator>
<dc:creator>Sanchez, E.</dc:creator>
<dc:creator>Shin, G. Y.</dc:creator>
<dc:creator>Spyer, M. J.</dc:creator>
<dc:creator>Walker, P. A.</dc:creator>
<dc:creator>Kjaer, S.</dc:creator>
<dc:creator>Riddell, A.</dc:creator>
<dc:creator>Beale, R.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Stockinger, B.</dc:creator>
<dc:creator>Gamblin, S.</dc:creator>
<dc:creator>McCoy, L. E.</dc:creator>
<dc:creator>Cherepanov, P.</dc:creator>
<dc:creator>Nastouli, E.</dc:creator>
<dc:creator>Kassiotis, G.</dc:creator>
<dc:date>2020-05-15</dc:date>
<dc:identifier>doi:10.1101/2020.05.14.095414</dc:identifier>
<dc:title><![CDATA[Pre-existing and de novo humoral immunity to SARS-CoV-2 in humans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.05.14.096115v1?rss=1">
<title>
<![CDATA[
Non-canonical autophagy drives alternative ATG8 conjugation to phosphatidylserine 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.05.14.096115v1?rss=1"
</link>
<description><![CDATA[
Autophagy is a fundamental catabolic process essential for development, homeostasis and proper immune function 1. During autophagy, a cascade of ATG proteins target intracellular cargoes for lysosomal degradation and recycling 2. This pathway utilises a unique post-translational modification, the conjugation of ATG8 proteins to phosphatidylethanolamine (PE) at autophagosomes, which modulates cargo selection and maturation. ATG8 lipidation also occurs during non-canonical autophagy, a parallel pathway involving Single Membrane ATG8 Conjugation (SMAC) to endolysosomal compartments, which plays a key role in phagocytosis and other processes 3. It has been widely assumed that SMAC involves the same lipidation of ATG8 to PE, but this has yet to be formally tested. Here, we show that ATG8 undergoes alternative lipidation to phosphatidylserine (PS) during non-canonical autophagy/SMAC. Using mass spectrometry, we find that activation of SMAC, by pharmacological agents 4,5, or during non-canonical autophagy processes such as LC3-associated phagocytosis 6,7 and Influenza A virus infection 8, induces the covalent conjugation of ATG8 to PS, as well as PE. This alternative lipidation event is dependent on the ATG16L1 WD40 domain, and occurs at PS enriched endolysosomal membranes. Importantly, we find that the ATG8-PS and ATG8-PE adducts are differentially delipidated by isoforms of the ATG4 family, indicating significant molecular distinctions and mechanisms between these two species.

Together, these results provide an important new insight into autophagy signalling, revealing an alternative form of the hallmark ATG8-lipidation event, so widely used to define and assay autophagy. Furthermore, ATG8-PS lipidation provides a specific  molecular signature for non-canonical autophagy, uncovering a novel means of detecting and monitoring this emerging pathway.
]]></description>
<dc:creator>Durgan, J.</dc:creator>
<dc:creator>Lystad, A. H.</dc:creator>
<dc:creator>Sloan, K.</dc:creator>
<dc:creator>Carlsson, S. R.</dc:creator>
<dc:creator>Wilson, M. I.</dc:creator>
<dc:creator>Marcassa, E.</dc:creator>
<dc:creator>Ulferts, R.</dc:creator>
<dc:creator>Webster, J.</dc:creator>
<dc:creator>Lopez-Clavijo, A. F.</dc:creator>
<dc:creator>Wakelam, M. J.</dc:creator>
<dc:creator>Beale, R.</dc:creator>
<dc:creator>Simonsen, A.</dc:creator>
<dc:creator>Oxley, D.</dc:creator>
<dc:creator>Florey, O.</dc:creator>
<dc:date>2020-05-15</dc:date>
<dc:identifier>doi:10.1101/2020.05.14.096115</dc:identifier>
<dc:title><![CDATA[Non-canonical autophagy drives alternative ATG8 conjugation to phosphatidylserine]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.05.18.097618v1?rss=1">
<title>
<![CDATA[
The PAXFOXO1s trigger fast trans-differentiation of chick embryonic neural cells into alveolar rhabdomyosarcoma with tissue invasive properties limited by S phase entry inhibition 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.05.18.097618v1?rss=1"
</link>
<description><![CDATA[
The chromosome translocations generating PAX3FOXO1 and PAX7FOXO1 chimeric proteins are the primary hallmarks of the paediatric cancer, Alveolar Rhabdomyosarcoma (ARMS). Despite the ability of these transcription factors to remodel chromatin landscapes and promote the expression of tumour driver genes, they only inefficiently promote malignant transformation in vivo. The reason for this is unclear. To address this, we developed an in ovo model to follow the response of spinal cord progenitors to PAXFOXO1s. Our data demonstrate that PAXFOXO1s, but not wild-type PAX3 and PAX7, trigger the trans-differentiation of neural cells into ARMS-like cells with myogenic characteristics. In parallel expression of PAXFOXO1s remodels the neural pseudo-stratified epithelium into a cohesive mesenchyme capable of tissue invasion. Surprisingly, gain for PAXFOXO1s, as for wild-type PAX3/7, reduces the levels of CDK-CYCLIN activity and arrests cells in G1. Introduction of CYCLIN D1 or MYCN overcomes PAXFOXO1s mediated cell cycle inhibition and promotes tumour growth. Together, our findings reveal a mechanism underpinning the apparent limited oncogenicity of PAXFOXO1 fusion transcription factors and support a neural origin for ARMS.
]]></description>
<dc:creator>GONZALEZ CURTO, G.</dc:creator>
<dc:creator>Der Vartanian, A.</dc:creator>
<dc:creator>FRARMA, Y.</dc:creator>
<dc:creator>Manceau, L.</dc:creator>
<dc:creator>Baldi, L.</dc:creator>
<dc:creator>Prisco, S.</dc:creator>
<dc:creator>ELAROUCI, N.</dc:creator>
<dc:creator>Causeret, F.</dc:creator>
<dc:creator>RIGOLET, M.</dc:creator>
<dc:creator>AURADE, F.</dc:creator>
<dc:creator>DE REYNIES, A.</dc:creator>
<dc:creator>CONTREMOULINS, V.</dc:creator>
<dc:creator>RELAIX, F.</dc:creator>
<dc:creator>FAKLARIS, O.</dc:creator>
<dc:creator>BRISCOE, J.</dc:creator>
<dc:creator>GILARDI-HEBENSTREIT, P.</dc:creator>
<dc:creator>RIBES, V.</dc:creator>
<dc:date>2020-05-20</dc:date>
<dc:identifier>doi:10.1101/2020.05.18.097618</dc:identifier>
<dc:title><![CDATA[The PAXFOXO1s trigger fast trans-differentiation of chick embryonic neural cells into alveolar rhabdomyosarcoma with tissue invasive properties limited by S phase entry inhibition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.05.23.108977v1?rss=1">
<title>
<![CDATA[
Frizzled-dependent Planar Cell Polarity without Wnt Ligands 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.05.23.108977v1?rss=1"
</link>
<description><![CDATA[
Planar cell polarity (PCP) organizes the orientation of cellular protrusions and migratory activity within the tissue plane. PCP establishment involves the subcellular polarization of core PCP components. It has been suggested Wnt gradients could provide a global cue that coordinates local PCP with tissue axes. Here we dissect the role of Wnt ligands in the orientation of hairs of Drosophila wings, an established system for study of PCP. We found that PCP was normal in quintuple mutant wings that rely solely on membrane-tethered Wingless for Wnt signaling, suggesting that a Wnt gradient is not required. We then used a nanobody-based approach to trap Wntless in the endoplasmic reticulum, and hence prevent all Wnt secretion, specifically during the period of PCP establishment. PCP was still established. We conclude that, even though Wnt ligands could contribute to PCP, they are not essential, and another global cue must exist for tissue-wide polarization.
]]></description>
<dc:creator>Yu, J. J. S.</dc:creator>
<dc:creator>Maugarny-Cales, A.</dc:creator>
<dc:creator>Pelletier, S.</dc:creator>
<dc:creator>Alexandre, C.</dc:creator>
<dc:creator>Bellaiche, Y.</dc:creator>
<dc:creator>Vincent, J.-P.</dc:creator>
<dc:creator>McGough, I. J.</dc:creator>
<dc:date>2020-05-24</dc:date>
<dc:identifier>doi:10.1101/2020.05.23.108977</dc:identifier>
<dc:title><![CDATA[Frizzled-dependent Planar Cell Polarity without Wnt Ligands]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.05.25.114504v1?rss=1">
<title>
<![CDATA[
FAM83F regulates canonical Wnt signalling through an interaction with CK1α 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.05.25.114504v1?rss=1"
</link>
<description><![CDATA[
The function of the FAM83F protein, like the functions of many members of the FAM83 family, is poorly understood. Here we show that injection of Fam83f mRNA into Xenopus embryos causes axis duplication, a phenotype indicative of enhanced Wnt signalling. Consistent with this, overexpression of FAM83F activates Wnt signalling, whilst ablation of FAM83F from human colorectal cancer (CRC) cells attenuates it. We demonstrate that FAM83F is farnesylated and interacts and co-localises with CK1 at the plasma membrane. This interaction with CK1 is essential for FAM83F to activate Wnt signalling, and FAM83F mutants that do not interact with CK1 fail to induce axis duplication in Xenopus embryos and to activate Wnt signalling in cells. FAM83F acts upstream of the {beta}-catenin destruction complex, because the attenuation of Wnt signalling caused by loss of FAM83F can be rescued by GSK-3 inhibition. Introduction of a farnesyl-deficient mutant mis-localises the FAM83F-CK1 complex to the nucleus and significantly attenuates Wnt signalling, indicating that FAM83F exerts its effects on Wnt signalling at the plasma membrane.
]]></description>
<dc:creator>Dunbar, K.</dc:creator>
<dc:creator>Jones, R. A.</dc:creator>
<dc:creator>Dingwell, K. S.</dc:creator>
<dc:creator>Macartney, T.</dc:creator>
<dc:creator>Smith, J. C.</dc:creator>
<dc:creator>Sapkota, G. P.</dc:creator>
<dc:date>2020-05-26</dc:date>
<dc:identifier>doi:10.1101/2020.05.25.114504</dc:identifier>
<dc:title><![CDATA[FAM83F regulates canonical Wnt signalling through an interaction with CK1α]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.05.29.123513v1?rss=1">
<title>
<![CDATA[
CD147 (BSG) but not ACE2 expression is detectable in vascular endothelial cells within single cell RNA sequencing datasets derived from multiple tissues in healthy individuals 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.05.29.123513v1?rss=1"
</link>
<description><![CDATA[
Coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and is associated with a wide range of systemic manifestations. Several observations support a role for vascular endothelial dysfunction in the pathogenesis including an increased incidence of thrombotic events and coagulopathy and the presence of vascular risk factors as an independent predictor of poor prognosis. It has recently been reported that endothelitis is associated with viral inclusion bodies suggesting a direct role for SARS-CoV-2 in the pathogenesis. The ACE2 receptor has been shown to mediate SARS-CoV-2 uptake and it has been proposed that CD147 (BSG) can function as an alternative cell surface receptor. To define the endothelial cell populations that are susceptible to infection with SARS-CoV-2, we investigated the expression of ACE2 as well as other genes implicated in the cellular entry of SARS-Cov-2 in the vascular endothelium through the analysis of single cell sequencing data derived from multiple human tissues (skin, liver, kidney, lung and intestine). We found that CD147 (BSG) but not ACE2 is detectable in vascular endothelial cells within single cell sequencing datasets derived from multiple tissues in healthy individuals. This implies that either ACE2 is not expressed in healthy tissue but is instead induced in response to SARS-Cov2 or that SARS-Cov2 enters endothelial cells via an alternative receptor such as CD147.
]]></description>
<dc:creator>Ganier, C.</dc:creator>
<dc:creator>Du-Harpur, X.</dc:creator>
<dc:creator>Harun, N.</dc:creator>
<dc:creator>Wan, B.</dc:creator>
<dc:creator>Arthurs, C.</dc:creator>
<dc:creator>Luscombe, N.</dc:creator>
<dc:creator>Watt, F.</dc:creator>
<dc:creator>Lynch, M.</dc:creator>
<dc:date>2020-05-31</dc:date>
<dc:identifier>doi:10.1101/2020.05.29.123513</dc:identifier>
<dc:title><![CDATA[CD147 (BSG) but not ACE2 expression is detectable in vascular endothelial cells within single cell RNA sequencing datasets derived from multiple tissues in healthy individuals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.03.131391v1?rss=1">
<title>
<![CDATA[
20 years of African Neuroscience: Waking a sleeping giant 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.03.131391v1?rss=1"
</link>
<description><![CDATA[
Understanding the function and dysfunction of the brain remains one of the key challenges of our time. However, an overwhelming majority of brain research is carried out in the Global North, by a minority of well-funded and intimately interconnected labs. In contrast, with an estimated one neuroscientist per million people in Africa, news about neuroscience research from the Global South remains sparse. Clearly, devising new policies to boost Africa’s neuroscience landscape is imperative. However, the policy must be based on accurate data, which is largely lacking. Such data must reflect the extreme heterogeneity of research outputs across the continent’s 54 countries distributed over an area larger than USA, Europe and China combined. Here, we analysed all of Africa’s Neuroscience output over the past 21 years. Uniquely, we individually verified in each of 12,326 publications that the work was indeed performed in Africa and led by African-based researchers. This step is critical: previous estimates grossly inflated figures, because many of Africa’s high-visibility publications are in fact the result of internationally led collaborations, with most work done outside of Africa. The remaining number of African-led Neuroscience publications was 5,219, on average only ~5 per country and year. From here, we extracted metrics such as the journal and citations, as well as detailed information on funding, international collaborations and the techniques and model systems used. We link these metrics to demographic data and indicators of mobility and economy. For reference, we also extracted the same metrics from 220 randomly selected publications each from the UK, USA, Australia, Japan and Brazil. Our unique dataset allows us to gain accurate and in-depth information on the current state of African Neuroscience research, and to put it into a global context. This in turn allows us to make actionable recommendations on how African research might best be supported in the future.Competing Interest StatementThe authors have declared no competing interest.View Full Text
]]></description>
<dc:creator>Maina, M. B.</dc:creator>
<dc:creator>Ahmad, U.</dc:creator>
<dc:creator>Harun, I. A.</dc:creator>
<dc:creator>Kwairanga, H. S.</dc:creator>
<dc:creator>Nasr, F. E.</dc:creator>
<dc:creator>Salihu, A. T.</dc:creator>
<dc:creator>Abushouk, A. I.</dc:creator>
<dc:creator>Muhammad, A.</dc:creator>
<dc:creator>Awadelkareem, M. A.</dc:creator>
<dc:creator>Amin, A.</dc:creator>
<dc:creator>Imam, A.</dc:creator>
<dc:creator>Akinrinade, I. D.</dc:creator>
<dc:creator>Yakubu, A. H.</dc:creator>
<dc:creator>Azeez, I. A.</dc:creator>
<dc:creator>Mohammed, Y. G.</dc:creator>
<dc:creator>Adamu, A. A.</dc:creator>
<dc:creator>Ibrahim, B. H.</dc:creator>
<dc:creator>Bukar, A. M.</dc:creator>
<dc:creator>Yaro, A. U.</dc:creator>
<dc:creator>Prieto-Godino, L. L.</dc:creator>
<dc:creator>Baden, T.</dc:creator>
<dc:date>2020-06-04</dc:date>
<dc:identifier>doi:10.1101/2020.06.03.131391</dc:identifier>
<dc:title><![CDATA[20 years of African Neuroscience: Waking a sleeping giant]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.04.133306v1?rss=1">
<title>
<![CDATA[
Systematic analysis of mutational spectra associated with DNA repair deficiency in C. elegans 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.04.133306v1?rss=1"
</link>
<description><![CDATA[
Genome integrity is particularly important in germ cells to faithfully preserve genetic information across generations. As yet little is known about the contribution of various DNA repair pathways to prevent mutagenesis. Using the C. elegans model we analyse mutational spectra that arise in wild-type and 61 DNA repair and DNA damage response mutants cultivated over multiple generations. Overall, 44% of lines show >2-fold increased mutagenesis with a broad spectrum of mutational outcomes including changes in single or multiple types of base substitutions induced by defects in base excision or nucleotide excision repair, or elevated levels of 50-400 bp deletions in translesion polymerase mutants rev-3(pol {zeta}) and polh-1(pol {eta}). Mutational signatures associated with defective homologous recombination fall into two classes: 1) mutants lacking brc-1/BRCA1 or rad-51/RAD51 paralogs show elevated base substitutions, indels and structural variants, while 2) deficiency for MUS-81/MUS81 and SLX-1/SLX1 nucleases, and HIM-6/BLM, HELQ-1/HELQ and RTEL-1/RTEL1 helicases primarily cause structural variants. Genome-wide investigation of mutagenesis patterns identified elevated rates of tandem duplications often associated with inverted repeats in helq-1 mutants, and a unique pattern of  translocation events involving homeologous sequences in rip-1 paralog mutants. atm-1/ATM DNA damage checkpoint mutants harboured complex structural variants enriched in subtelomeric regions, and chromosome end-to-end fusions. Finally, while inactivation of the p53-like gene cep-1 did not affect mutagenesis, combined brc-1 cep-1 deficiency displayed increased, locally clustered mutagenesis. In summary, we provide a global view of how DNA repair pathways prevent germ cell mutagenesis.
]]></description>
<dc:creator>Meier, B.</dc:creator>
<dc:creator>Volkova, N. N.</dc:creator>
<dc:creator>Hong, Y.</dc:creator>
<dc:creator>Bertolini, S.</dc:creator>
<dc:creator>Gonzalez-Huici, V.</dc:creator>
<dc:creator>Petrova, T.</dc:creator>
<dc:creator>Boulton, S.</dc:creator>
<dc:creator>Campbell, P. J.</dc:creator>
<dc:creator>Gerstung, M.</dc:creator>
<dc:creator>Gartner, A.</dc:creator>
<dc:date>2020-06-04</dc:date>
<dc:identifier>doi:10.1101/2020.06.04.133306</dc:identifier>
<dc:title><![CDATA[Systematic analysis of mutational spectra associated with DNA repair deficiency in C. elegans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.05.135913v1?rss=1">
<title>
<![CDATA[
Frequent loss-of-heterozygosity in CRISPR-Cas9-edited early human embryos 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.05.135913v1?rss=1"
</link>
<description><![CDATA[
CRISPR-Cas9 genome editing is a promising technique for clinical applications, such as the correction of disease-associated alleles in somatic cells. The use of this approach has also been discussed in the context of heritable editing of the human germline. However, studies assessing gene correction in early human embryos report low efficiency of mutation repair, high rates of mosaicism and the possibility of unintended editing outcomes that may have pathologic consequences. We developed computational pipelines to assess single-cell genomics and transcriptomics datasets from OCT4 (POU5F1) CRISPR-Cas9-targeted and control human preimplantation embryos. This allowed us to evaluate on-target mutations that would be missed by more conventional genotyping techniques. We observed loss-of-heterozygosity in edited cells that spanned regions beyond the POU5F1 on-target locus, as well as segmental loss and gain of chromosome 6, on which the POU5F1 gene is located. Unintended genome editing outcomes were present in approximately 16% of the human embryo cells analysed and spanned 4 to 20kb. Our observations are consistent with recent findings indicating complexity at on-target sites following CRISPR-Cas9 genome editing. Our work underscores the importance of further basic research to assess the safety of genome editing techniques in human embryos, which will inform debates about the potential clinical use of this technology.
]]></description>
<dc:creator>Alanis-Lobato, G.</dc:creator>
<dc:creator>Zohren, J.</dc:creator>
<dc:creator>McCarthy, A.</dc:creator>
<dc:creator>Fogarty, N. M. E.</dc:creator>
<dc:creator>Kubikova, N.</dc:creator>
<dc:creator>Hardman, E.</dc:creator>
<dc:creator>Greco, M.</dc:creator>
<dc:creator>Wells, D.</dc:creator>
<dc:creator>Turner, J. M. A.</dc:creator>
<dc:creator>Niakan, K.</dc:creator>
<dc:date>2020-06-05</dc:date>
<dc:identifier>doi:10.1101/2020.06.05.135913</dc:identifier>
<dc:title><![CDATA[Frequent loss-of-heterozygosity in CRISPR-Cas9-edited early human embryos]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.08.139808v1?rss=1">
<title>
<![CDATA[
Repurposing the orphan drug nitisinone to control the transmission of African trypanosomiasis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.08.139808v1?rss=1"
</link>
<description><![CDATA[
Tsetse transmit African trypanosomiasis, which is a disease fatal to both humans and animals. A vaccine to protect against this disease does not exist so transmission control relies on eliminating tsetse populations. Although neurotoxic insecticides are the gold standard for insect control, they negatively impact the environment and reduce insect pollinator species. Here we present a promising, environment-friendly alternative that targets insect tyrosine metabolism pathway. A bloodmeal contains high levels of tyrosine, which is toxic to haematophagous insects if it is not degraded. RNAi silencing of either the first two enzymes in the tyrosine degradation pathway (TAT and HPPD) was lethal to tsetse. Furthermore, nitisinone (NTBC), an FDA-approved tyrosine catabolism inhibitor, killed tsetse regardless if the drug was orally or topically applied. However, it did not affect bumblebee survival. A mathematical model shows that NTBC could reduce the transmission of African trypanosomiasis in sub-Saharan Africa, thus accelerating current elimination programmes.
]]></description>
<dc:creator>Acosta-Serrano, A.</dc:creator>
<dc:creator>Sterkel, M.</dc:creator>
<dc:creator>Haines, L.</dc:creator>
<dc:creator>Casas-Sanchez, A.</dc:creator>
<dc:creator>Owino, V.</dc:creator>
<dc:creator>Vionette-Amaral, R.</dc:creator>
<dc:creator>Quek, S.</dc:creator>
<dc:creator>Rose, C.</dc:creator>
<dc:creator>Silva dos Santos, M.</dc:creator>
<dc:creator>Garcia Escude, N.</dc:creator>
<dc:creator>Ismail, H.</dc:creator>
<dc:creator>Paine, M. I.</dc:creator>
<dc:creator>Barribeau, S. M.</dc:creator>
<dc:creator>Wagstaff, S.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Masiga, D.</dc:creator>
<dc:creator>Yakob, L.</dc:creator>
<dc:creator>Oliveira, P. L.</dc:creator>
<dc:date>2020-06-08</dc:date>
<dc:identifier>doi:10.1101/2020.06.08.139808</dc:identifier>
<dc:title><![CDATA[Repurposing the orphan drug nitisinone to control the transmission of African trypanosomiasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.08.140228v1?rss=1">
<title>
<![CDATA[
An optogenetic method for interrogating YAP1 and TAZ nuclear-cytoplasmic shuttling 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.08.140228v1?rss=1"
</link>
<description><![CDATA[
The shuttling of transcription factors and transcriptional regulators in and out of the nucleus is central to the regulation of many biological processes. Here we describe a new method for studying the rates of nuclear entry and exit of transcriptional regulators. A photo-responsive AsLOV (Avena sativa Light Oxygen Voltage) domain is used to sequester fluorescently-labelled transcriptional regulators YAP1 and TAZ/WWTR1 on the surface of mitochondria and reversibly release them upon blue light illumination. After dissociation, fluorescent signals from mitochondria, cytoplasm and nucleus are extracted with a bespoke app and used to generate rates of nuclear entry and exit. Using this method, we demonstrate that phosphorylation of YAP1 on canonical sites enhances its rate of nuclear export. Moreover, we provide evidences that, despite high intercellular variability, YAP1 import and export rates correlated within the same cell. By simultaneously releasing YAP1 and TAZ from sequestration, we show that their rates of entry and exit are correlated. Furthermore, tracking of light-sensitive YAP1 with lattice lightsheet microscopy revealed high heterogeneity of YAP1 dynamics within different subcellular regions, suggesting that implementing high resolution volumetric 3D data could shed light on new mechanisms of nuclear-cytoplasmic shuttling of proteins.
]]></description>
<dc:creator>Dowbaj, A. M.</dc:creator>
<dc:creator>Jenkins, R. P.</dc:creator>
<dc:creator>Hahn, K.</dc:creator>
<dc:creator>Montagner, M.</dc:creator>
<dc:creator>Sahai, E. M.</dc:creator>
<dc:date>2020-06-09</dc:date>
<dc:identifier>doi:10.1101/2020.06.08.140228</dc:identifier>
<dc:title><![CDATA[An optogenetic method for interrogating YAP1 and TAZ nuclear-cytoplasmic shuttling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.09.142638v1?rss=1">
<title>
<![CDATA[
SOX2 is required independently in both stem and differentiated cells for pituitary tumour genesis in p27 null mice. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.09.142638v1?rss=1"
</link>
<description><![CDATA[
Loss of P27 predominantly results in development of murine pituitary intermediate lobe (IL) tumours. We previously showed that the pleiotropic protein P27 can drive repression of the transcription factor Sox2. This interaction plays an important role during development of p27-/- IL tumours because loss of one copy of Sox2 diminishes tumorigenesis. Here, we have explored the cellular origin and mechanisms underlying melanotroph tumorigenesis in p27-/- IL. We show that IL hyperplasia is associated with reduced cellular differentiation, while levels of SOX2 increase in both stem cells (SC) and melanotrophs. Using loss-of-function and lineage tracing approaches, we demonstrate that SOX2 is required cell-autonomously in p27-/- melanotrophs and SCs for tumorigenesis. This is supported by studies deleting the Sox2 regulatory region 2 (Srr2), which is the target of P27 repressive action. Single cell transcriptomic analysis reveals that activation of a SOX2-dependent MAPK pathway in SCs is important for p27-/- tumorigenesis. Our data highlight different roles of SOX2 following loss of p27, according to the cellular context. Furthermore, we uncover a tumor-promoting function for SCs, which is SOX2-dependant. In conclusion, our results imply that targeting SCs, in addition to tumour cells themselves, may represent an efficient anti-tumoral strategy in certain contexts.
]]></description>
<dc:creator>Moncho-Amor, V.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Galichet, C.</dc:creator>
<dc:creator>Matheu, A.</dc:creator>
<dc:creator>Lovell-Badge, R.</dc:creator>
<dc:creator>Rizzoti, K.</dc:creator>
<dc:date>2020-06-10</dc:date>
<dc:identifier>doi:10.1101/2020.06.09.142638</dc:identifier>
<dc:title><![CDATA[SOX2 is required independently in both stem and differentiated cells for pituitary tumour genesis in p27 null mice.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.11.147017v1?rss=1">
<title>
<![CDATA[
Critical role of WNK1 in MYC-dependent early thymocyte development 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.11.147017v1?rss=1"
</link>
<description><![CDATA[
WNK1, a kinase that controls kidney salt homeostasis, also regulates adhesion and migration in CD4+ T cells. Wnk1 is highly expressed in thymocytes, and since migration is important for thymocyte maturation, we investigated a role for WNK1 in thymocyte development. We find that WNK1 is required for the transition of double negative (DN) thymocytes through the {beta}-selection checkpoint and subsequent proliferation and differentiation into double positive (DP) thymocytes. Furthermore, we show that WNK1 negatively regulates LFA1-mediated adhesion and positively regulates CXCL12-induced migration in DN thymocytes. Despite this, migration defects of WNK1-deficient thymocytes do not account for the developmental arrest. Instead, we show that in DN thymocytes WNK1 transduces pre-TCR signals via OXSR1 and STK39 kinases and the SLC12A2 ion co-transporter that are required for post-transcriptional upregulation of MYC and subsequent proliferation and differentiation into DP thymocytes. Thus, a pathway regulating ion homeostasis is a critical regulator of thymocyte development.
]]></description>
<dc:creator>Köchl, R.</dc:creator>
<dc:creator>Vanes, L.</dc:creator>
<dc:creator>Llorian Sopena, M.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Hartweger, H.</dc:creator>
<dc:creator>Fountain, K.</dc:creator>
<dc:creator>White, A.</dc:creator>
<dc:creator>Cowan, J.</dc:creator>
<dc:creator>Anderson, G.</dc:creator>
<dc:creator>Tybulewicz, V.</dc:creator>
<dc:date>2020-06-12</dc:date>
<dc:identifier>doi:10.1101/2020.06.11.147017</dc:identifier>
<dc:title><![CDATA[Critical role of WNK1 in MYC-dependent early thymocyte development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.11.145672v1?rss=1">
<title>
<![CDATA[
Dentate gyrus development requires a cortical hem-derived astrocytic scaffold. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.11.145672v1?rss=1"
</link>
<description><![CDATA[
During embryonic development, radial glial cells give rise to neurons, then to astrocytes following the gliogenic switch. Timely regulation of the switch, operated by several transcription factors, is fundamental for allowing coordinated interactions between neurons and glia. We deleted the gene for one such factor, SOX9, early during mouse brain development and observed a significantly compromised dentate gyrus (DG). We dissected the origin of the defect, targeting embryonic Sox9 deletion to either the DG neuronal progenitor domain or the adjacent cortical hem (CH). We identified in the latter previously uncharacterized ALDH1L1+ astrocytic progenitors, which form a fimbrial-specific glial scaffold necessary for neuronal progenitor migration towards the developing DG. Our results highlight an early crucial role of SOX9 for DG development through regulation of astroglial potential acquisition in the CH. Moreover, we illustrate how formation of a local network, amidst astrocytic and neuronal progenitors originating from adjacent domains, underlays brain morphogenesis.
]]></description>
<dc:creator>Caramello, A.</dc:creator>
<dc:creator>Galichet, C.</dc:creator>
<dc:creator>Lovell-Badge, R.</dc:creator>
<dc:creator>Rizzoti, K.</dc:creator>
<dc:date>2020-06-12</dc:date>
<dc:identifier>doi:10.1101/2020.06.11.145672</dc:identifier>
<dc:title><![CDATA[Dentate gyrus development requires a cortical hem-derived astrocytic scaffold.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.12.147983v1?rss=1">
<title>
<![CDATA[
Pan-cancer detection of driver genes at the single-patient resolution 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.12.147983v1?rss=1"
</link>
<description><![CDATA[
BackgroundIdentifying the complete repertoire of genes that drive cancer in individual patients is crucial for precision oncology. Most established methods identify driver genes that are recurrently altered across patient cohorts. However, mapping these genes back to patients leaves a sizeable fraction with few or no drivers, hindering our understanding of cancer mechanisms and limiting the choice of therapeutic interventions.

ResultsWe present sysSVM2, a machine learning software that integrates cancer genetic alterations with gene systems-level properties to predict drivers in individual patients. Using simulated pan-cancer data, we optimise sysSVM2 for application to any cancer type. We benchmark its performance on real cancer data and validate its applicability to a rare cancer type with few known driver genes. We show that drivers predicted by sysSVM2 have a low false-positive rate, are stable and disrupt well-known cancer-related pathways.

ConclusionssysSVM2 can be used to identify driver alterations in patients lacking sufficient canonical drivers or belonging to rare cancer types for which assembling a large enough cohort is challenging, furthering the goals of precision oncology. As resources for the community, we provide the code to implement sysSVM2 and the pre-trained models in all TCGA cancer types (https://github.com/ciccalab/sysSVM2).
]]></description>
<dc:creator>Nulsen, J.</dc:creator>
<dc:creator>Misetic, H.</dc:creator>
<dc:creator>Yau, C.</dc:creator>
<dc:creator>Ciccarelli, F. D.</dc:creator>
<dc:date>2020-06-12</dc:date>
<dc:identifier>doi:10.1101/2020.06.12.147983</dc:identifier>
<dc:title><![CDATA[Pan-cancer detection of driver genes at the single-patient resolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.12.149534v1?rss=1">
<title>
<![CDATA[
Structure of the cysteine-rich domain of Plasmodium falciparum P113 identifies the location of the RH5 binding site 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.12.149534v1?rss=1"
</link>
<description><![CDATA[
Plasmodium falciparum RH5 is a secreted parasite ligand that is essential for erythrocyte invasion through direct interaction with the host erythrocyte receptor basigin. RH5 forms a tripartite complex with two other secreted parasite proteins: CyRPA and RIPR, and is tethered to the surface of the parasite through membrane-anchored P113. Antibodies against RH5, CyRPA and RIPR inhibit parasite invasion, suggesting that vaccines containing these three components have the potential to prevent blood-stage malaria. To further explore the role of the P113-RH5 interaction, we selected monoclonal antibodies against P113 that were either inhibitory or non-inhibitory for RH5 binding. Using a Fab fragment as a crystallisation chaperone, we determined the crystal structure of the RH5-binding region of P113 and showed that it is composed of two domains with structural similarities to rhamnose-binding lectins. We identified the RH5 binding site on P113 by using a combination of hydrogen-deuterium exchange mass spectrometry and site directed mutagenesis. We found that a monoclonal antibody to P113 that bound to this interface and inhibited the RH5-P113 interaction did not inhibit parasite blood-stage growth. These findings provide further structural information on the protein interactions of RH5 and will be helpful in the development of blood-stage malaria vaccines that target RH5.

ImportanceMalaria is a deadly infectious disease primarily caused by the parasite Plasmodium falciparum. It remains a major global health problem and there is no highly effective vaccine. A parasite protein called RH5 is centrally involved in the invasion of host red blood cells, making it - and the other parasite proteins it interacts with - promising vaccine targets. We recently identified a protein called P113 that binds RH5 suggesting that it anchors RH5 to the parasite surface. In this paper, we use structural biology to locate and characterize the RH5 binding region on P113. These findings will be important to guide the development of new anti-malarial vaccines to ultimately prevent this disease which affects some of the poorest people on the planet.
]]></description>
<dc:creator>Campeotto, I.</dc:creator>
<dc:creator>Galaway, F.</dc:creator>
<dc:creator>Mehmood, S.</dc:creator>
<dc:creator>Barfod, L.</dc:creator>
<dc:creator>Quinkert, D.</dc:creator>
<dc:creator>Kotraiah, V.</dc:creator>
<dc:creator>Phares, T. W.</dc:creator>
<dc:creator>Wright, K. E.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Draper, S. J.</dc:creator>
<dc:creator>Higgins, M.</dc:creator>
<dc:creator>Wright, G. J.</dc:creator>
<dc:date>2020-06-13</dc:date>
<dc:identifier>doi:10.1101/2020.06.12.149534</dc:identifier>
<dc:title><![CDATA[Structure of the cysteine-rich domain of Plasmodium falciparum P113 identifies the location of the RH5 binding site]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.12.145151v1?rss=1">
<title>
<![CDATA[
Integrating genotypes and phenotypes improves long-term forecasts of seasonal influenza A/H3N2 evolution 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.12.145151v1?rss=1"
</link>
<description><![CDATA[
Seasonal influenza virus A/H3N2 is a major cause of death globally. Vaccination remains the most effective preventative. Rapid mutation of hemagglutinin allows viruses to escape adaptive immunity. This antigenic drift necessitates regular vaccine updates. Effective vaccine strains need to represent H3N2 populations circulating one year after strain selection. Experts select strains based on experimental measurements of antigenic drift and predictions made by models from hemagglutinin sequences. We developed a novel influenza forecasting framework that integrates phenotypic measures of antigenic drift and functional constraint with previously published sequence-only fitness estimates. Forecasts informed by phenotypic measures of antigenic drift consistently outperformed previous sequence-only estimates, while sequence-only estimates of functional constraint surpassed more comprehensive experimentally-informed estimates. Importantly, the best models integrated estimates of both functional constraint and either antigenic drift phenotypes or recent population growth.
]]></description>
<dc:creator>Huddleston, J.</dc:creator>
<dc:creator>Barnes, J. R.</dc:creator>
<dc:creator>Rowe, T.</dc:creator>
<dc:creator>Kondor, R.</dc:creator>
<dc:creator>Wentworth, D. E.</dc:creator>
<dc:creator>Whittaker, L.</dc:creator>
<dc:creator>Ermetal, B.</dc:creator>
<dc:creator>Daniels, R. S.</dc:creator>
<dc:creator>McCauley, J. W.</dc:creator>
<dc:creator>Fujisaki, S.</dc:creator>
<dc:creator>Nakamura, K.</dc:creator>
<dc:creator>Kishida, N.</dc:creator>
<dc:creator>Watanabe, S.</dc:creator>
<dc:creator>Hasegawa, H.</dc:creator>
<dc:creator>Barr, I.</dc:creator>
<dc:creator>Subbarao, K.</dc:creator>
<dc:creator>Neher, R.</dc:creator>
<dc:creator>Bedford, T.</dc:creator>
<dc:date>2020-06-13</dc:date>
<dc:identifier>doi:10.1101/2020.06.12.145151</dc:identifier>
<dc:title><![CDATA[Integrating genotypes and phenotypes improves long-term forecasts of seasonal influenza A/H3N2 evolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.13.149716v1?rss=1">
<title>
<![CDATA[
Phase separation induced by cohesin SMC protein complexes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.13.149716v1?rss=1"
</link>
<description><![CDATA[
Cohesin is a key protein complex that organizes the spatial structure of chromosomes during interphase. Here, we show that yeast cohesin shows pronounced clustering on DNA in an ATP-independent manner, exhibiting all the hallmarks of phase separation. In vitro visualization of cohesin on DNA shows DNA-cohesin clusters that exhibit liquid-like behavior. This includes mutual fusion and reversible dissociation upon depleting the cohesin concentration, increasing the ionic strength, or adding 1,6-hexanediol, conditions that disrupt weak interactions. We discuss how bridging-induced phase separation can explain the DNA-cohesin clustering through DNA-cohesin-DNA bridges. We confirm that, in vivo, a fraction of cohesin associates with chromatin in yeast cells in a manner consistent with phase separation. Our findings establish that SMC proteins can exhibit phase separation, which has potential to clarify previously unexplained aspects of in vivo SMC behavior and constitute an additional principle by which SMC complexes impact genome organization.

One sentence summaryYeast cohesin complex is observed to phase separate with DNA into liquid droplets, which it accomplishes by ATP-independent DNA bridging.
]]></description>
<dc:creator>Ryu, J.-K.</dc:creator>
<dc:creator>Bouchoux, C.</dc:creator>
<dc:creator>Liu, H. W.</dc:creator>
<dc:creator>Kim, E.</dc:creator>
<dc:creator>Minamino, M.</dc:creator>
<dc:creator>de Groot, R.</dc:creator>
<dc:creator>Katan, A. J.</dc:creator>
<dc:creator>Bonato, A.</dc:creator>
<dc:creator>Marenduzzo, D.</dc:creator>
<dc:creator>Michieletto, D.</dc:creator>
<dc:creator>Uhlmann, F.</dc:creator>
<dc:creator>Dekker, C.</dc:creator>
<dc:date>2020-06-14</dc:date>
<dc:identifier>doi:10.1101/2020.06.13.149716</dc:identifier>
<dc:title><![CDATA[Phase separation induced by cohesin SMC protein complexes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.12.147033v1?rss=1">
<title>
<![CDATA[
Outcomes of the 2019 EMDataResource model challenge: validation of cryo-EM models at near-atomic resolution 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.12.147033v1?rss=1"
</link>
<description><![CDATA[
This paper describes outcomes of the 2019 Cryo-EM Map-based Model Metrics Challenge sponsored by EMDataResource (www.emdataresource.org). The goals of this challenge were (1) to assess the quality of models that can be produced using current modeling software, (2) to check the reproducibility of modeling results from different software developers and users, and (3) compare the performance of current metrics used for evaluation of models. The focus was on near-atomic resolution maps with an innovative twist: three of four target maps formed a resolution series (1.8 to 3.1 [A]) from the same specimen and imaging experiment. Tools developed in previous challenges were expanded for managing, visualizing and analyzing the 63 submitted coordinate models, and several novel metrics were introduced. The results permit specific recommendations to be made about validating near-atomic cryo-EM structures both in the context of individual laboratory experiments and holdings of structure data archives such as the Protein Data Bank. Our findings demonstrate the relatively high accuracy and reproducibility of cryo-EM models derived from these benchmark maps by 13 participating teams, representing both widely used and novel modeling approaches. We also evaluate the pros and cons of the commonly used metrics to assess model quality and recommend the adoption of multiple scoring parameters to provide full and objective annotation and assessment of the model, reflective of the observed density in the cryo-EM map.
]]></description>
<dc:creator>Lawson, C. L.</dc:creator>
<dc:creator>Kryshtafovych, A.</dc:creator>
<dc:creator>Adams, P. D.</dc:creator>
<dc:creator>Afonine, P.</dc:creator>
<dc:creator>Baker, M. L.</dc:creator>
<dc:creator>Barad, B. A.</dc:creator>
<dc:creator>Bond, P.</dc:creator>
<dc:creator>Burnley, T.</dc:creator>
<dc:creator>Cao, R.</dc:creator>
<dc:creator>Cheng, J.</dc:creator>
<dc:creator>Chojnowski, G.</dc:creator>
<dc:creator>Cowtan, K.</dc:creator>
<dc:creator>Dill, K. A.</dc:creator>
<dc:creator>DiMaio, F.</dc:creator>
<dc:creator>Farrell, D.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:creator>Herzik, M. A.</dc:creator>
<dc:creator>Hoh, S. W.</dc:creator>
<dc:creator>Hou, J.</dc:creator>
<dc:creator>Hung, L.-W.</dc:creator>
<dc:creator>Igaev, M.</dc:creator>
<dc:creator>Joseph, A. P.</dc:creator>
<dc:creator>Kihara, D.</dc:creator>
<dc:creator>Kumar, D.</dc:creator>
<dc:creator>Mittal, S.</dc:creator>
<dc:creator>Monastyrskyy, B.</dc:creator>
<dc:creator>Olek, M.</dc:creator>
<dc:creator>Palmer, C.</dc:creator>
<dc:creator>Patwardhan, A.</dc:creator>
<dc:creator>Perez, A.</dc:creator>
<dc:creator>Pfab, J.</dc:creator>
<dc:creator>Pintilie, G. D.</dc:creator>
<dc:creator>Richardson, J. S.</dc:creator>
<dc:creator>Rosenthal, P. B.</dc:creator>
<dc:creator>Sarkar, D.</dc:creator>
<dc:creator>Schaefer, L. U.</dc:creator>
<dc:creator>Schmid, M. F.</dc:creator>
<dc:creator>Schroeder, G. F.</dc:creator>
<dc:creator>Shekhar, M.</dc:creator>
<dc:creator>Si, D.</dc:creator>
<dc:creator>Singharoy, A.</dc:creator>
<dc:creator>Terashi, G.</dc:creator>
<dc:creator>Terwilliger, T. C.</dc:creator>
<dc:creator>Vaiana, A.</dc:creator>
<dc:creator>Wan</dc:creator>
<dc:date>2020-06-15</dc:date>
<dc:identifier>doi:10.1101/2020.06.12.147033</dc:identifier>
<dc:title><![CDATA[Outcomes of the 2019 EMDataResource model challenge: validation of cryo-EM models at near-atomic resolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.16.155564v1?rss=1">
<title>
<![CDATA[
Rostrocaudal Patterning and Neural Crest Differentiation of Human Pre-Neural Spinal Cord Progenitors in vitro 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.16.155564v1?rss=1"
</link>
<description><![CDATA[
The spinal cord emerges from a niche of neuromesodermal progenitors (NMPs) formed and maintained by Wnt/FGF signals at the posterior end of the embryo. NMPs can be generated from human pluripotent stem cells and hold promise for spinal cord replacement therapies. However, NMPs are transient, which complicates the full range production of rostrocaudal spinal cord identities in vitro. Here we report the generation of NMP-derived pre-neural progenitors (PNPs) with stem cell-like self-renewal capacity. PNPs maintain pre-spinal cord identity by co-expressing the transcription factors SOX2 and CDX2, and lose mesodermal potential by downregulating TBXT. For 7 to 10 passages PNPs divide to self-renew and to make trunk neural crest (NC), while gradually adopting a more posterior identity by activating colinear HOX gene expression. This HOX clock can be halted at the thoracic level for up to 30 passages by blocking the trunk-to-tail transition through GDF11-mediated signal inhibition.
]]></description>
<dc:creator>Cooper, F.</dc:creator>
<dc:creator>Gentsch, G. E.</dc:creator>
<dc:creator>Mitter, R.</dc:creator>
<dc:creator>Bouissou, C.</dc:creator>
<dc:creator>Healy, L.</dc:creator>
<dc:creator>Hernandez-Rodriguez, A.</dc:creator>
<dc:creator>Smith, J. C.</dc:creator>
<dc:creator>Bernardo, A. S.</dc:creator>
<dc:date>2020-06-16</dc:date>
<dc:identifier>doi:10.1101/2020.06.16.155564</dc:identifier>
<dc:title><![CDATA[Rostrocaudal Patterning and Neural Crest Differentiation of Human Pre-Neural Spinal Cord Progenitors in vitro]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.10.144402v1?rss=1">
<title>
<![CDATA[
Human Whole Genome Sequencing in South Africa 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.10.144402v1?rss=1"
</link>
<description><![CDATA[
The advent and evolution of next generation sequencing has considerably impacted genomic research. Until recently, South African researchers were unable to access affordable platforms capable of human whole genome sequencing locally and DNA samples had to be exported.  Here we report the whole genome sequences of the first six human DNA samples sequenced and analysed at the South African Medical Research Council Genomics Centre. We demonstrate that the data obtained is of high quality, with an average sequencing depth of 36.41, and that the output is comparable to data generated internationally on a similar platform. The Genomics Centre creates an environment where African researchers are able to access world class facilities, increasing local capacity to sequence whole genomes as well as store and analyse the data.Competing Interest StatementThe authors have declared no competing interest.View Full Text
]]></description>
<dc:creator>Glanzmann, B.</dc:creator>
<dc:creator>Jooste, T.</dc:creator>
<dc:creator>Ghoor, S.</dc:creator>
<dc:creator>Gordon, R.</dc:creator>
<dc:creator>Mia, R.</dc:creator>
<dc:creator>Mao, R.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Charls, P.</dc:creator>
<dc:creator>Douman, C.</dc:creator>
<dc:creator>Kotze, M.</dc:creator>
<dc:creator>Peeters, A.</dc:creator>
<dc:creator>Loots, G.</dc:creator>
<dc:creator>Esser, M.</dc:creator>
<dc:creator>Tiemessen, C.</dc:creator>
<dc:creator>Wilkinson, R.</dc:creator>
<dc:creator>Louw, J.</dc:creator>
<dc:creator>Gray, G.</dc:creator>
<dc:creator>Warren, R.</dc:creator>
<dc:creator>Moller Moller, M.</dc:creator>
<dc:creator>Kinnear, C.</dc:creator>
<dc:date>2020-06-10</dc:date>
<dc:identifier>doi:10.1101/2020.06.10.144402</dc:identifier>
<dc:title><![CDATA[Human Whole Genome Sequencing in South Africa]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.18.159103v1?rss=1">
<title>
<![CDATA[
Maladaptive nutrient signalling sustains the m.3243A>G mtDNA mutation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.18.159103v1?rss=1"
</link>
<description><![CDATA[
Mutations of the mitochondrial genome (mtDNA) cause a range of profoundly debilitating clinical conditions for which treatment options are very limited. Most mtDNA diseases show heteroplasmy - tissues express both wild-type and mutant mtDNA. While the level of heteroplasmy broadly correlates with disease severity, the relationships between specific mtDNA mutations, heteroplasmy, disease phenotype and severity are poorly understood. We have carried out extensive bioenergetic, metabolomic and RNAseq studies on heteroplasmic patient derived cells carrying the most prevalent disease related mtDNA mutation, m.3243A>G. These studies reveal that the mutation promotes changes in metabolites which is associated with the upregulation of the PI3K-Akt-mTORC1 axis in patient-derived cells and tissues. Remarkably, pharmacological inhibition of PI3K, Akt, or mTORC1 activated mitophagy, reduced mtDNA mutant load and rescued cellular bioenergetic function. The rescue was prevented by inhibition of mitophagy. The PI3K-Akt-mTORC1 axis thus represents a potential therapeutic target that may benefit people suffering from the consequences of the m.3243A>G mutation.
]]></description>
<dc:creator>Chung, C.-Y.</dc:creator>
<dc:creator>Singh, K.</dc:creator>
<dc:creator>Kotiadis, V. N.</dc:creator>
<dc:creator>Ahn, J. H.</dc:creator>
<dc:creator>Kabir, L.</dc:creator>
<dc:creator>Topley, E.</dc:creator>
<dc:creator>Tan, J.</dc:creator>
<dc:creator>Andrews, W. D.</dc:creator>
<dc:creator>Plotegher, N.</dc:creator>
<dc:creator>Guella, G.</dc:creator>
<dc:creator>Pitceathly, R. D. S.</dc:creator>
<dc:creator>Szabadkai, G.</dc:creator>
<dc:creator>Yuneva, M.</dc:creator>
<dc:creator>Duchen, M. R.</dc:creator>
<dc:date>2020-06-19</dc:date>
<dc:identifier>doi:10.1101/2020.06.18.159103</dc:identifier>
<dc:title><![CDATA[Maladaptive nutrient signalling sustains the m.3243A>G mtDNA mutation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.19.162115v1?rss=1">
<title>
<![CDATA[
Genetic dissection of Down syndrome-associated alterations in APP/amyloid-β biology using mouse models 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.19.162115v1?rss=1"
</link>
<description><![CDATA[
Individuals who have Down syndrome (caused by trisomy of chromosome 21), have a greatly elevated risk of early-onset Alzheimers disease, in which amyloid-{beta} accumulates in the brain. Amyloid-{beta} is a product of the chromosome 21 gene APP (amyloid precursor protein) and the extra copy or  dose of APP is thought to be the cause of this early-onset Alzheimers disease. However, other chromosome 21 genes likely modulate disease when in three-copies in people with Down syndrome. Here we show that an extra copy of chromosome 21 genes, other than APP, influences APP/A{beta} biology. We crossed Down syndrome mouse models with partial trisomies, to an APP transgenic model and found that extra copies of subgroups of chromosome 21 gene(s) modulate amyloid-{beta} aggregation and APP transgene-associated mortality, independently of changing amyloid precursor protein abundance. Thus, genes on chromosome 21, other than APP, likely modulate Alzheimers disease in people who have Down syndrome.
]]></description>
<dc:creator>Tosh, J. L.</dc:creator>
<dc:creator>Rhymes, E.</dc:creator>
<dc:creator>Mumford, P.</dc:creator>
<dc:creator>Whittaker, H. T.</dc:creator>
<dc:creator>Pulford, L. J.</dc:creator>
<dc:creator>Noy, S. J.</dc:creator>
<dc:creator>Cleverley, K.</dc:creator>
<dc:creator>Walker, M. L.</dc:creator>
<dc:creator>Tybulewicz, V. L. J.</dc:creator>
<dc:creator>Wykes, R. C.</dc:creator>
<dc:creator>Fisher, E. M. C.</dc:creator>
<dc:creator>Wiseman, F. K.</dc:creator>
<dc:date>2020-06-20</dc:date>
<dc:identifier>doi:10.1101/2020.06.19.162115</dc:identifier>
<dc:title><![CDATA[Genetic dissection of Down syndrome-associated alterations in APP/amyloid-β biology using mouse models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.19.161513v1?rss=1">
<title>
<![CDATA[
CRYO-EM STRUCTURE OF THE DELTA-RETROVIRAL INTASOME IN COMPLEX WITH THE PP2A REGULATORY SUBUNIT B56gamma 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.19.161513v1?rss=1"
</link>
<description><![CDATA[
The Retroviridae delta-retrovirus genus includes the most oncogenic pathogen- human T-cell lymphotropic virus type 1 (HTLV-1)(1). Many of the ~20 million people infected with HTLV-1 will develop severe leukaemia (2) or an ALS-like motor disease (3) unless a therapy becomes available. A key step in the establishment of infection is the integration of viral genetic material into the host genome, catalysed by the viral integrase (IN) enzyme. Here we used X-ray crystallography and single-particle cryo-electron microscopy to determine the structure of functional delta-retroviral IN assembled on viral DNA ends and bound the B56{gamma} subunit of its human host factor, the protein phosphatase 2A (4). The structure reveals a tetrameric IN assembly bound to the phosphatase via a conserved short linear motif found within the extended linker connecting the catalytic core (CCD) and C-terminal (CTD) IN domains. Unexpectedly, all four IN subunits are involved in B56{gamma} binding, taking advantage of the flexibility of the CCD-CTD linkers. Our results fill the current gap in the structural understanding of the delta-retroviral integration machinery. Insight into the interactions between the delta-retroviral intasome and the host will be crucial for understanding the pattern of integration events in infected individuals and therefore bears important clinical implications.
]]></description>
<dc:creator>Barski, M. S.</dc:creator>
<dc:creator>Minnell, J. J.</dc:creator>
<dc:creator>Hodakova, Z.</dc:creator>
<dc:creator>Pye, V.</dc:creator>
<dc:creator>Nans, A.</dc:creator>
<dc:creator>Cherepanov, P.</dc:creator>
<dc:creator>Maertens, G. N.</dc:creator>
<dc:date>2020-06-20</dc:date>
<dc:identifier>doi:10.1101/2020.06.19.161513</dc:identifier>
<dc:title><![CDATA[CRYO-EM STRUCTURE OF THE DELTA-RETROVIRAL INTASOME IN COMPLEX WITH THE PP2A REGULATORY SUBUNIT B56gamma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.22.166215v1?rss=1">
<title>
<![CDATA[
From local resynchronization to global pattern recovery in the zebrafish segmentation clock 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.22.166215v1?rss=1"
</link>
<description><![CDATA[
ABSTRACTRhythmic spatial gene expression patterns termed the segmentation clock regulate vertebrate body axis segmentation during embryogenesis. The integrity of these patterns requires local synchronization between neighboring cells by Delta-Notch signaling and its inhibition results in defective segment boundaries. The oscillating tissue deforms substantially throughout development, but whether such tissue-scale morphogenesis complements local synchronization during pattern generation and segment formation is not understood. Here, we investigate pattern recovery in the zebrafish segmentation clock by washing out a Notch inhibitor, allowing resynchronization at different developmental stages, and analyzing the recovery of normal segments. Although from previous work no defects are expected after recovery, we find that washing out at early stages causes a distinctive intermingling of normal and defective segments, suggesting unexpectedly large fluctuations of synchrony before complete recovery. To investigate this recovery behavior, we develop a new model of the segmentation clock combining key ingredients motivated by prior experimental observations: coupling between neighboring oscillators, a frequency profile, a gradient of cell mixing, tissue length change, and cell advection pattern. This model captures the experimental observation of intermingled normal and defective segments through the formation of persistent phase vortices of the genetic oscillators. Experimentally observed recovery patterns at different developmental stages are predicted by temporal changes of tissue-level properties, such as tissue length and cell advection pattern in the model. These results suggest that segmental pattern recovery occurs at two scales: local pattern formation and transport of these patterns through tissue morphogenesis, highlighting a generic mechanism of pattern dynamics within developing tissues.SIGNIFICANCE Interacting genetic oscillators can generate a coherent rhythm and a tissue-level pattern from an initially desynchronized state. Using experiment and theory we study resynchronization and pattern recovery of the zebrafish segmentation clock, which makes the embryonic body segments. Experimental perturbation of intercellular signaling with an inhibitor results in intermingled normal and defective segments. According to theory, this behavior may be caused by persistent local vortices scattered in the tissue during pattern recovery. Full pattern recovery follows dynamic global properties, such as tissue length and advection pattern, in contrast to other genetic oscillators in a static tissue such as circadian clocks. Our work highlights how dynamics of tissue level properties may couple to biochemical pattern formation in tissues and developing embryos.Competing Interest StatementThe authors have declared no competing interest.View Full Text
]]></description>
<dc:creator>Uriu, K.</dc:creator>
<dc:creator>Liao, B.-K.</dc:creator>
<dc:creator>Oates, A.</dc:creator>
<dc:creator>Morelli, L.</dc:creator>
<dc:date>2020-06-23</dc:date>
<dc:identifier>doi:10.1101/2020.06.22.166215</dc:identifier>
<dc:title><![CDATA[From local resynchronization to global pattern recovery in the zebrafish segmentation clock]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.24.168567v1?rss=1">
<title>
<![CDATA[
The native structure of the full-length, assembled influenza A virus matrix protein, M1. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.24.168567v1?rss=1"
</link>
<description><![CDATA[
Influenza A virus causes millions of severe illnesses during annual epidemics. The most abundant protein in influenza virions is the matrix protein M1 that mediates virus assembly by forming an endoskeleton beneath the virus membrane. The structure of full-length M1, and how it oligomerizes to mediate assembly of virions, is unknown. Here we have determined the complete structure of assembled M1 within intact virus particles, as well as the structure of M1 oligomers reconstituted in vitro. We found that the C-terminal domain of M1 is disordered in solution, but can fold and bind in trans to the N-terminal domain of another M1 monomer, thus polymerising M1 into linear strands which coat the interior surface of the assembling virion membrane. In the M1 polymer, five histidine residues, contributed by three different M1 monomers, form a cluster that can serve as the pH-sensitive disassembly switch after entry into a target cell. These structures therefore provide mechanisms for influenza virus assembly and disassembly.
]]></description>
<dc:creator>Peukes, J.</dc:creator>
<dc:creator>Xiong, X.</dc:creator>
<dc:creator>Erlendsson, S.</dc:creator>
<dc:creator>Qu, K.</dc:creator>
<dc:creator>Wan, W.</dc:creator>
<dc:creator>Calder, L. J.</dc:creator>
<dc:creator>Schraidt, O.</dc:creator>
<dc:creator>Kummer, S.</dc:creator>
<dc:creator>Freund, S. M. V.</dc:creator>
<dc:creator>Kräusslich, H.-G.</dc:creator>
<dc:creator>Briggs, J. A. G.</dc:creator>
<dc:date>2020-06-24</dc:date>
<dc:identifier>doi:10.1101/2020.06.24.168567</dc:identifier>
<dc:title><![CDATA[The native structure of the full-length, assembled influenza A virus matrix protein, M1.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.24.169193v1?rss=1">
<title>
<![CDATA[
The phosphatidylinositol 3-phosphate binding protein SNX4 controls ATG9A recycling and autophagy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.24.169193v1?rss=1"
</link>
<description><![CDATA[
Late endosomes and lysosomes (endolysosomes) receive proteins and cargo from the secretory, endocytic and autophagic pathways. Whereas these pathways and the degradative processes of endolysosomes are well characterized, less is understood about protein traffic from these organelles. In this study, we demonstrate the direct involvement of the phosphatidylinositol 3-phosphate (PI3P) binding SNX4 protein in membrane protein recycling from endolysosomes, and show that SNX4 is required for proper autophagic flux. We show that SNX4 mediates recycling of the transmembrane autophagy machinery protein ATG9A from endolysosomes to early endosomes, from where ATG9A is recycled to the trans-Golgi network in a retromer-dependent manner. Upon siRNA-mediated depletion of SNX4 or the retromer component VPS35, we observed accumulation of ATG9A on endolysosomes and early endosomes, respectively. Moreover, starvation-induced autophagosome biogenesis and autophagic flux were inhibited when SNX4 was downregulated. Altogether, we propose that proper ATG9A recycling by SNX4 sustains autophagy by preventing exhaustion of the available ATG9A pool.
]]></description>
<dc:creator>Stenmark, H.</dc:creator>
<dc:creator>Ravussin, A.</dc:creator>
<dc:creator>Tooze, S.</dc:creator>
<dc:date>2020-06-24</dc:date>
<dc:identifier>doi:10.1101/2020.06.24.169193</dc:identifier>
<dc:title><![CDATA[The phosphatidylinositol 3-phosphate binding protein SNX4 controls ATG9A recycling and autophagy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.23.166173v1?rss=1">
<title>
<![CDATA[
Centromere-specific antibody-mediated karyotyping of Okinawan Oikopleura dioica suggests the presence of three chromosomes. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.23.166173v1?rss=1"
</link>
<description><![CDATA[
Oikopleura dioica is a ubiquitous marine tunicate of biological interest due to features that include dioecious reproduction, short life cycle, and vertebrate-like dorsal notochord while possessing a relatively compact genome. The use of tunicates as model organisms, particularly with these characteristics, offers the advantage of facilitating studies in evolutionary development and furthering understanding of enduring attributes found in the more complex vertebrates. At present, we are undertaking an initiative to sequence the genomes of Oikopleura individuals in populations found among the seas surrounding the Ryukyu Islands in southern Japan. To facilitate and validate genome assemblies, karyotyping was employed to count individual animals chromosomes in situ using centromere-specific antibodies directed against H3S28P, a prophase-metaphase cell cycle-specific marker of histone H3. New imaging data of embryos and oocytes stained with two different antibodies were obtained; interpretation of these data lead us to conclude that the Okinawan Oikopleura dioica has three pairs of chromosomes, akin to previous results from genomic assemblies in Atlantic populations. The imaging data have been deposited to the open-access EBI BioImage Archive for reuse while additionally providing representative images of two commercially available anti-H3S28P antibodies staining properties for use in epifluorescent and confocal based fluorescent microscopy.
]]></description>
<dc:creator>Liu, A. W.</dc:creator>
<dc:creator>Tan, Y.</dc:creator>
<dc:creator>Masunaga, A.</dc:creator>
<dc:creator>Plessy, C.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:date>2020-06-24</dc:date>
<dc:identifier>doi:10.1101/2020.06.23.166173</dc:identifier>
<dc:title><![CDATA[Centromere-specific antibody-mediated karyotyping of Okinawan Oikopleura dioica suggests the presence of three chromosomes.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.24.167049v1?rss=1">
<title>
<![CDATA[
SARS-CoV-2 infection and replication in human fetal and pediatric gastric organoids 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.24.167049v1?rss=1"
</link>
<description><![CDATA[
Coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is a global public health emergency. COVID-19 typically manifests as a respiratory illness but an increasing number of clinical reports describe gastrointestinal (GI) symptoms. This is particularly true in children in whom GI symptoms are frequent and viral shedding outlasts viral clearance from the respiratory system. By contrast, fetuses seem to be rarely affected by COVID-19, although the virus has been detected in placentas of affected women. These observations raise the question of whether the virus can infect and replicate within the stomach once ingested. Moreover, it is not yet clear whether active replication of SARS-CoV-2 is possible in the stomach of children or in fetuses at different developmental stages. Here we show the novel derivation of fetal gastric organoids from 8-21 post-conception week (PCW) fetuses, and from pediatric biopsies, to be used as an in vitro model for SARS-CoV-2 gastric infection. Gastric organoids recapitulate human stomach with linear increase of gastric mucin 5AC along developmental stages, and expression of gastric markers pepsinogen, somatostatin, gastrin and chromogranin A. In order to investigate SARS-CoV-2 infection with minimal perturbation and under steady-state conditions, we induced a reversed polarity in the gastric organoids (RP-GOs) in suspension. In this condition of exposed apical polarity, the virus can easily access viral receptor angiotensin-converting enzyme 2 (ACE2). The pediatric RP-GOs are fully susceptible to infection with SARS-CoV-2, where viral nucleoprotein is expressed in cells undergoing programmed cell death, while the efficiency of infection is significantly lower in fetal organoids. The RP-GOs derived from pediatric patients show sustained robust viral replication of SARS-CoV-2, compared with organoids derived from fetal stomachs. Transcriptomic analysis shows a moderate innate antiviral response and the lack of differentially expressed genes belonging to the interferon family. Collectively, we established the first expandable human gastric organoid culture across fetal developmental stages, and we support the hypothesis that fetal tissue seems to be less susceptible to SARS-CoV-2 infection, especially in early stages of development. However, the virus can efficiently infect gastric epithelium in pediatric patients, suggesting that the stomach might have an active role in fecal-oral transmission of SARS-CoV-2.
]]></description>
<dc:creator>Giobbe, G. G.</dc:creator>
<dc:creator>Bonfante, F.</dc:creator>
<dc:creator>Zambaiti, E.</dc:creator>
<dc:creator>Gagliano, O.</dc:creator>
<dc:creator>Jones, B. C.</dc:creator>
<dc:creator>Luni, C.</dc:creator>
<dc:creator>Laterza, C.</dc:creator>
<dc:creator>Perin, S.</dc:creator>
<dc:creator>Stuart, H. T.</dc:creator>
<dc:creator>Pagliari, M.</dc:creator>
<dc:creator>Bortolami, A.</dc:creator>
<dc:creator>Mazzetto, E.</dc:creator>
<dc:creator>Manfredi, A.</dc:creator>
<dc:creator>Colantuono, C.</dc:creator>
<dc:creator>Di Filippo, L.</dc:creator>
<dc:creator>Pellegata, A. F.</dc:creator>
<dc:creator>Li, V. S. W.</dc:creator>
<dc:creator>Eaton, S.</dc:creator>
<dc:creator>Thapar, N.</dc:creator>
<dc:creator>Cacchiarelli, D.</dc:creator>
<dc:creator>Elvassore, N.</dc:creator>
<dc:creator>De Coppi, P.</dc:creator>
<dc:date>2020-06-24</dc:date>
<dc:identifier>doi:10.1101/2020.06.24.167049</dc:identifier>
<dc:title><![CDATA[SARS-CoV-2 infection and replication in human fetal and pediatric gastric organoids]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.15.152835v1?rss=1">
<title>
<![CDATA[
A thermostable, closed, SARS-CoV-2 spike protein trimer. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.15.152835v1?rss=1"
</link>
<description><![CDATA[
The spike (S) protein of SARS-CoV-2 mediates receptor binding and cell entry and is the dominant target of the immune system. S exhibits substantial conformational flexibility. It transitions from closed to open conformations to expose its receptor binding site, and subsequently from prefusion to postfusion conformations to mediate fusion of viral and cellular membranes. S protein derivatives are components of vaccine candidates and diagnostic assays, as well as tools for research into the biology and immunology of SARS-CoV-2. Here we have designed mutations in S which allow production of thermostable, crosslinked, S protein trimers that are trapped in the closed, pre-fusion, state. We have determined the structures of crosslinked and non-crosslinked proteins, identifying two distinct closed conformations of the S trimer. We demonstrate that the designed, thermostable, closed S trimer can be used in serological assays. This protein has potential applications as a reagent for serology, virology and as an immunogen.
]]></description>
<dc:creator>Xiong, X.</dc:creator>
<dc:creator>Qu, K.</dc:creator>
<dc:creator>Ciazynska, K. A.</dc:creator>
<dc:creator>Hosmillo, M.</dc:creator>
<dc:creator>Carter, A. P.</dc:creator>
<dc:creator>Ebrahimi, S.</dc:creator>
<dc:creator>Ke, Z.</dc:creator>
<dc:creator>Scheres, S. H. W.</dc:creator>
<dc:creator>Bergamaschi, L.</dc:creator>
<dc:creator>Grice, G. L.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>The CITIID-NIHR COVID-19 BioResource Collaboration,</dc:creator>
<dc:creator>Nathan, J. A.</dc:creator>
<dc:creator>Baker, S.</dc:creator>
<dc:creator>James, L. C.</dc:creator>
<dc:creator>Baxendale, H. E.</dc:creator>
<dc:creator>Goodfellow, I.</dc:creator>
<dc:creator>Doffinger, R.</dc:creator>
<dc:creator>Briggs, J. A. G.</dc:creator>
<dc:date>2020-06-17</dc:date>
<dc:identifier>doi:10.1101/2020.06.15.152835</dc:identifier>
<dc:title><![CDATA[A thermostable, closed, SARS-CoV-2 spike protein trimer.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.28.176040v1?rss=1">
<title>
<![CDATA[
Molecular basis for substrate specificity of the Phactr1/PP1 phosphatase holoenzyme 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.28.176040v1?rss=1"
</link>
<description><![CDATA[
ABSTRACTPPP-family phosphatases such as PP1 have little intrinsic specificity. Cofactors can target PP1 to substrates or subcellular locations, but it remains unclear how they might confer sequence-specificity on PP1. The cytoskeletal regulator Phactr1 is a neuronally-enriched PP1 cofactor that is controlled by G-actin. Structural analysis showed that Phactr1 binding remodels PP1’s hydrophobic groove, creating a new composite surface adjacent to the catalytic site. Using phosphoproteomics, we identified numerous fibroblast and neuronal Phactr1/PP1 substrates, which include cytoskeletal components and regulators. We determined high-resolution structures of Phactr1/PP1 bound to the dephosphorylated forms of its substrates IRSp53 and spectrin αII. Inversion of the phosphate in these holoenzyme-product complexes supports the proposed PPP-family catalytic mechanism. Substrate sequences C-terminal to the dephosphorylation site make intimate contacts with the composite Phactr1/PP1 surface, which are required for efficient dephosphorylation. Sequence specificity explains why Phactr1/PP1 exhibits orders-of-magnitude enhanced reactivity towards its substrates, compared to apo-PP1 or other PP1 holoenzymes.Competing Interest StatementThe authors have declared no competing interest.View Full Text
]]></description>
<dc:creator>Fedoryshchak, R. O.</dc:creator>
<dc:creator>Prechova, M.</dc:creator>
<dc:creator>Butler, A.</dc:creator>
<dc:creator>Lee, R.</dc:creator>
<dc:creator>O'Reilly, N.</dc:creator>
<dc:creator>Flynn, H.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Eder, N.</dc:creator>
<dc:creator>Ultanir, S.</dc:creator>
<dc:creator>Mouilleron, S.</dc:creator>
<dc:creator>Treisman, R.</dc:creator>
<dc:date>2020-06-28</dc:date>
<dc:identifier>doi:10.1101/2020.06.28.176040</dc:identifier>
<dc:title><![CDATA[Molecular basis for substrate specificity of the Phactr1/PP1 phosphatase holoenzyme]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.28.174821v1?rss=1">
<title>
<![CDATA[
Age-related degeneration leads to gliosis but not regeneration in the zebrafish retina 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.28.174821v1?rss=1"
</link>
<description><![CDATA[
Ageing is a significant risk factor for degeneration of the retina. Harnessing the regenerative potential of Muller glia cells (MG) in the retina offers great promise for the treatment of blinding conditions. Yet, the impact of ageing on MG regenerative capacity has not yet been considered. Here we show that the zebrafish retina undergoes telomerase-independent age-related neurodegeneration. Yet, this progressive neuronal loss in the ageing retina is insufficient to stimulate the MG regenerative response. Instead, age-related neurodegeneration leads to MG gliosis and loss of vision, similarly to humans. Nevertheless, gliotic MG cells retain Yap expression and the ability to regenerate neurons after acute light damage. Therefore, we identify key differences in the MG response to acute versus chronic damage in the zebrafish retina and show that aged gliotic MG can be stimulated to repair damaged neurons in old age.

SUMMARY

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=103 SRC="FIGDIR/small/174821v2_ufig1.gif" ALT="Figure 1">
View larger version (41K):
org.highwire.dtl.DTLVardef@1a184daorg.highwire.dtl.DTLVardef@1193750org.highwire.dtl.DTLVardef@7e65c0org.highwire.dtl.DTLVardef@4a2fe3_HPS_FORMAT_FIGEXP  M_FIG C_FIG Our data suggest there are key differences between mechanisms driving regeneration in response to acute damage versus age-related chronic damage. It may be that either the number of cells dying in natural ageing is not enough to stimulate MG to proliferate, or the low number of microglia and respective signals released are not sufficient to trigger MG proliferation. Importantly, we show that gliotic MG cells can be stimulated to repair damaged neurons in old zebrafish retina.
]]></description>
<dc:creator>Martins, R. R.</dc:creator>
<dc:creator>Zamzam, M.</dc:creator>
<dc:creator>Moosajee, M.</dc:creator>
<dc:creator>Thummel, R. B.</dc:creator>
<dc:creator>Henriques, C. M.</dc:creator>
<dc:creator>MacDonald, R. B.</dc:creator>
<dc:date>2020-06-29</dc:date>
<dc:identifier>doi:10.1101/2020.06.28.174821</dc:identifier>
<dc:title><![CDATA[Age-related degeneration leads to gliosis but not regeneration in the zebrafish retina]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.01.182709v1?rss=1">
<title>
<![CDATA[
Genetic architecture of host proteins interacting with SARS-CoV-2 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.01.182709v1?rss=1"
</link>
<description><![CDATA[
Strategies to develop therapeutics for SARS-CoV-2 infection may be informed by experimental identification of viral-host protein interactions in cellular assays and measurement of host response proteins in COVID-19 patients. Identification of genetic variants that influence the level or activity of these proteins in the host could enable rapid  in silico assessment in human genetic studies of their causal relevance as molecular targets for new or repurposed drugs to treat COVID-19. We integrated large-scale genomic and aptamer-based plasma proteomic data from 10,708 individuals to characterize the genetic architecture of 179 host proteins reported to interact with SARS-CoV-2 proteins or to participate in the host response to COVID-19. We identified 220 host DNA sequence variants acting in cis (MAF 0.01-49.9%) and explaining 0.3-70.9% of the variance of 97 of these proteins, including 45 with no previously known protein quantitative trait loci (pQTL) and 38 encoding current drug targets. Systematic characterization of pQTLs across the phenome identified protein-drug-disease links, evidence that putative viral interaction partners such as MARK3 affect immune response, and establish the first link between a recently reported variant for respiratory failure of COVID-19 patients at the ABO locus and hypercoagulation, i.e. maladaptive host response. Our results accelerate the evaluation and prioritization of new drug development programmes and repurposing of trials to prevent, treat or reduce adverse outcomes. Rapid sharing and dynamic and detailed interrogation of results is facilitated through an interactive webserver (https://omicscience.org/apps/covidpgwas/).
]]></description>
<dc:creator>Pietzner, M.</dc:creator>
<dc:creator>Wheeler, E.</dc:creator>
<dc:creator>Carrasco-Zanini, J.</dc:creator>
<dc:creator>Raffler, J.</dc:creator>
<dc:creator>Kerrison, N. D.</dc:creator>
<dc:creator>Oerton, E.</dc:creator>
<dc:creator>Auyeung, V. P. W.</dc:creator>
<dc:creator>Luan, J.</dc:creator>
<dc:creator>Finan, C.</dc:creator>
<dc:creator>Casas, J. P.</dc:creator>
<dc:creator>Ostroff, R.</dc:creator>
<dc:creator>Williams, S. A.</dc:creator>
<dc:creator>Kastenmüller, G.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:creator>Gamazon, E. G.</dc:creator>
<dc:creator>Wareham, N. J.</dc:creator>
<dc:creator>Hingorani, A. D.</dc:creator>
<dc:creator>Langenberg, C.</dc:creator>
<dc:date>2020-07-01</dc:date>
<dc:identifier>doi:10.1101/2020.07.01.182709</dc:identifier>
<dc:title><![CDATA[Genetic architecture of host proteins interacting with SARS-CoV-2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.01.182774v1?rss=1">
<title>
<![CDATA[
A 3-hydroxyacyl-CoA dehydratase (DEH) involved in very long chain fatty acid synthesis is crucial for oocyst mitotic division but not cell size during parasite transmission in Plasmodium 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.01.182774v1?rss=1"
</link>
<description><![CDATA[
Cells use fatty acids (FAs) for membrane biosynthesis, energy storage and the generation of signaling molecules. 3-hydroxyacyl-CoA dehydratase - DEH - is a key component of very long chain FA (VLCFA) synthesis. Here, we further characterized in-depth the location and function of DEH, applying in silico analysis, live cell imaging, reverse genetics and ultrastructure analysis using the mouse malaria model Plasmodium berghei. DEH is evolutionarily conserved across eukaryotes, with a single DEH in Plasmodium spp. and up to three orthologs in the other eukaryotes studied. DEH-GFP live-cell imaging showed strong GFP fluorescence throughout the life-cycle, with areas of localized expression in the cytoplasm and a circular ring pattern around the nucleus that colocalized with ER markers. {Delta}deh mutants showed a small but significant reduction in oocyst size compared to WT controls from day 10 post-infection onwards and endomitotic cell division and sporogony were completely ablated, blocking parasite transmission from mosquito to vertebrate host. Ultrastructure analysis confirmed degeneration of {Delta}deh oocysts, and a complete lack of sporozoite budding. Overall, DEH is evolutionarily conserved, localizes to the ER and plays a crucial role in sporogony.
]]></description>
<dc:creator>Guttery, D. S.</dc:creator>
<dc:creator>Pandey, R.</dc:creator>
<dc:creator>Ferguson, D.</dc:creator>
<dc:creator>Wall, R.</dc:creator>
<dc:creator>Brady, D.</dc:creator>
<dc:creator>Gupta, D.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:creator>Tewari, R.</dc:creator>
<dc:date>2020-07-02</dc:date>
<dc:identifier>doi:10.1101/2020.07.01.182774</dc:identifier>
<dc:title><![CDATA[A 3-hydroxyacyl-CoA dehydratase (DEH) involved in very long chain fatty acid synthesis is crucial for oocyst mitotic division but not cell size during parasite transmission in Plasmodium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.02.183871v1?rss=1">
<title>
<![CDATA[
High-resolution structures of malaria parasite actomyosin and actin filaments 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.02.183871v1?rss=1"
</link>
<description><![CDATA[
Malaria is responsible for half a million deaths annually and poses a huge economic burden on the developing world. The mosquito-borne parasites (Plasmodium spp.) that cause the disease depend upon an unconventional actomyosin motor for both gliding motility and host cell invasion. The motor system, often referred to as the glideosome complex, remains to be understood in molecular terms and is an attractive target for new drugs that might block the infection pathway. Here, we present the first high-resolution structure of the actomyosin motor complex from Plasmodium falciparum. Our structure includes the malaria parasite actin filament (PfAct1) complexed with the myosin motor (PfMyoA) and its two associated light-chains. The high-resolution core structure reveals the PfAct1:PfMyoA interface in atomic detail, while at lower-resolution, we visualize the PfMyoA light-chain binding region, including the essential light chain (PfELC) and the myosin tail interacting protein (PfMTIP). Finally, we report a bare PfAct1 filament structure at an improved resolution, which gives new information about the nucleotide-binding site, including the orientation of the ATP/ADP sensor, Ser15, and the presence of a channel, which we propose as a possible phosphate exit path after ATP hydrolysis.Significance statement We present the first structure of the malaria parasite motor complex; actin 1 (PfAct1) and myosin A (PfMyoA) with its two light chains. We also report a high-resolution structure of filamentous PfAct1 that reveals new atomic details of the ATPase site, including a channel, which may provide an exit route for phosphate and explain why phosphate release is faster in PfAct1 compared to canonical actins. PfAct1 goes through no conformational changes upon PfMyoA binding. Our PfMyoA structure also superimposes with a recent crystal structure of PfMyoA alone, though there are small but important conformational changes at the interface. Our structures serve as an excellent starting point for drug design against malaria, which is one of the most devastating infectious diseases.Competing Interest StatementThe authors have declared no competing interest.View Full Text
]]></description>
<dc:creator>Vahokoski, J.</dc:creator>
<dc:creator>Calder, L. J.</dc:creator>
<dc:creator>Lopez, A. J.</dc:creator>
<dc:creator>Molloy, J. E.</dc:creator>
<dc:creator>Rosenthal, P. B.</dc:creator>
<dc:creator>Kursula, I.</dc:creator>
<dc:date>2020-07-02</dc:date>
<dc:identifier>doi:10.1101/2020.07.02.183871</dc:identifier>
<dc:title><![CDATA[High-resolution structures of malaria parasite actomyosin and actin filaments]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.03.186031v1?rss=1">
<title>
<![CDATA[
Plasmodium berghei kinesin-5 associates with the spindle apparatus during cell division but is dispensable for parasite proliferation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.03.186031v1?rss=1"
</link>
<description><![CDATA[
Kinesin-5 motors play essential roles in spindle apparatus assembly during cell division, by generating forces to establish and maintain the spindle bipolarity essential for proper chromosome segregation. Kinesin-5 is largely conserved structurally and functionally in model eukaryotes, but its role is unknown in the Plasmodium parasite, an evolutionarily divergent organism with several atypical features of both mitotic and meiotic cell division. We have investigated the function and subcellular location of kinesin-5 during cell division throughout the Plasmodium berghei life cycle. Deletion of kinesin-5 had little visible effect at any proliferative stage except sporozoite production in oocysts, resulting in a significant decrease in the number of motile sporozoites in mosquito salivary glands, which were able to infect a new vertebrate host. Live-cell imaging showed kinesin-5-GFP located on the spindle and at spindle poles during both atypical mitosis and meiosis. Fixed-cell immunofluorescence assays revealed kinesin-5 co-localized with -tubulin and centrin-2 and a partial overlap with kinetochore marker NDC80 during early blood stage schizogony. Dual-colour live-cell imaging showed that kinesin-5 is closely associated with NDC80 during male gametogony, but not with kinesin-8B, a marker of the basal body and axonemes of the forming flagella. Treatment of gametocytes with microtubule-specific inhibitors confirmed kinesin-5 association with nuclear spindles and not cytoplasmic axonemal microtubules. Altogether, our results demonstrate that kinesin-5 is associated with the spindle apparatus, expressed in proliferating parasite stages, and important for efficient production of infectious sporozoites.
]]></description>
<dc:creator>Zeeshan, M.</dc:creator>
<dc:creator>Brady, D.</dc:creator>
<dc:creator>Stanway, R. R.</dc:creator>
<dc:creator>Moores, C.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:creator>Tewari, R.</dc:creator>
<dc:date>2020-07-03</dc:date>
<dc:identifier>doi:10.1101/2020.07.03.186031</dc:identifier>
<dc:title><![CDATA[Plasmodium berghei kinesin-5 associates with the spindle apparatus during cell division but is dispensable for parasite proliferation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.02.185744v1?rss=1">
<title>
<![CDATA[
Macrophage-specific responses to human- and animal-adapted tubercle bacilli reveal pathogen and host factors driving multinucleated cell formation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.02.185744v1?rss=1"
</link>
<description><![CDATA[
The Mycobacterium tuberculosis complex (MTBC) is a group of related pathogens that cause tuberculosis (TB) in mammals. MTBC species are distinguished by their ability to sustain in distinct host populations. While Mycobacterium bovis (Mbv) sustains transmission cycles in cattle and wild animals and causes zoonotic TB, M. tuberculosis (Mtb) affects human populations and seldom causes disease in cattle. However, the host and pathogen determinants driving host tropism between MTBC species are still unknown. Macrophages are the main host cell that encounters mycobacteria upon initial infection and we hypothesised that early interactions between the macrophage and mycobacteria influence species-specific disease outcome. To identify factors that contribute to host tropism, we analysed both blood-derived primary human and bovine macrophages (hM{phi} or bM{phi}, respectively) infected with Mbv and Mtb. We show that Mbv and Mtb reside in different cellular compartments and differentially replicate in hM{phi} whereas both Mbv and Mtb efficiently replicate in bM{phi}. Specifically, we show that out of the four infection combinations, only the infection of bM{phi} with Mbv promoted the formation of multinucleated cells (MNCs), a hallmark of tuberculous granulomas. Mechanistically, we demonstrate that both MPB70 from Mbv and extracellular vesicles released by Mbv-infected bM{phi} promote macrophage multi-nucleation. Importantly, we extend our in vitro studies to show that granulomas from Mbv-infected but not Mtb-infected cattle contained higher numbers of MNCs. Our findings implicate MNC formation in the contrasting pathology between Mtb and Mbv for the bovine host, and identify MPB70 from Mbv and extracellular vesicles from bM{phi} as mediators of this process.
]]></description>
<dc:creator>Queval, C. J.</dc:creator>
<dc:creator>Fearns, A.</dc:creator>
<dc:creator>Botella, L.</dc:creator>
<dc:creator>Smyth, A.</dc:creator>
<dc:creator>Schnettger, L.</dc:creator>
<dc:creator>Mitermite, M.</dc:creator>
<dc:creator>Wooff, E.</dc:creator>
<dc:creator>Villareal-Ramos, B.</dc:creator>
<dc:creator>Garcia-Gimenez, W.</dc:creator>
<dc:creator>Heunis, T.</dc:creator>
<dc:creator>Trost, M.</dc:creator>
<dc:creator>Werling, D.</dc:creator>
<dc:creator>Salguero, F. J.</dc:creator>
<dc:creator>Gordon, S. V.</dc:creator>
<dc:creator>Gutierrez, M.</dc:creator>
<dc:date>2020-07-03</dc:date>
<dc:identifier>doi:10.1101/2020.07.02.185744</dc:identifier>
<dc:title><![CDATA[Macrophage-specific responses to human- and animal-adapted tubercle bacilli reveal pathogen and host factors driving multinucleated cell formation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.09.195347v1?rss=1">
<title>
<![CDATA[
PRC1 and EB1 Binding Dynamics Reveal a Solidifying Central Spindle during Anaphase Compaction in Human Cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.09.195347v1?rss=1"
</link>
<description><![CDATA[
During mitosis the spindle undergoes morphological and dynamic changes. It reorganizes at the onset of anaphase when the antiparallel bundler PRC1 accumulates and recruits central spindle proteins to the midzone. Little is known about how the dynamic properties of the central spindle change during its morphological changes in human cells. Using gene editing, we generated human cells that express from their endogenous locus fluorescent PRC1 and EB1 to quantify their native spindle distribution and binding/unbinding turnover. EB1 plus end tracking revealed a general slowdown of microtubule growth, while PRC1, similar to its yeast orthologue Ase1, binds increasingly strongly to compacting antiparallel microtubule overlaps. KIF4A and CLASP1 bind more dynamically to the central spindle, but also show slowing down turnover. These results show that the central spindle gradually becomes more stable during mitosis, in agreement with a recent  bundling, sliding and compaction model of antiparallel midzone bundle formation in the central spindle during late mitosis.
]]></description>
<dc:creator>Jayant Asthana</dc:creator>
<dc:creator>Nicholas I. Cade</dc:creator>
<dc:creator>Wei Ming Lim</dc:creator>
<dc:creator>Thomas Surrey</dc:creator>
<dc:date>2020-07-09</dc:date>
<dc:identifier>doi:10.1101/2020.07.09.195347</dc:identifier>
<dc:title><![CDATA[PRC1 and EB1 Binding Dynamics Reveal a Solidifying Central Spindle during Anaphase Compaction in Human Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.12.198994v1?rss=1">
<title>
<![CDATA[
Ventricular, atrial and outflow tract heart progenitors arise from spatially and molecularly distinct regions of the primitive streak. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.12.198994v1?rss=1"
</link>
<description><![CDATA[
The heart develops from two sources of mesoderm progenitors, the first and second heart field (FHF and SHF). Using a single cell transcriptomic assay in combination with genetic lineage tracing, we find the FHF and SHF are subdivided into distinct pools of progenitors in gastrulating mouse embryos at earlier stages than previously thought. Each subpopulation has a distinct origin in the primitive streak. The first progenitors to leave the primitive streak contribute to the left ventricle, shortly after right ventricle progenitor emigrate, followed by the outflow tract and atrial progenitors. Moreover, a subset of atrial progenitors are gradually incorporated in posterior locations of the FHF. Although cells allocated to the outflow tract and atrium leave the primitive streak at a similar stage, they arise from different regions. Outflow tract cells originate from distal locations in the primitive streak while atrial progenitors are positioned more proximally. Moreover, single cell RNA sequencing demonstrates that the primitive streak cells contributing to the ventricles have a distinct molecular signature from those forming the outflow tract and atrium. We conclude that cardiac progenitors are pre-patterned within the primitive streak and this prefigures their allocation to distinct anatomical structures of the heart. Together, our data provide a new molecular and spatial map of mammalian cardiac progenitors that will support future studies of heart development, function and disease.
]]></description>
<dc:creator>Kenzo Ivanovitch</dc:creator>
<dc:creator>Pablo Soro-Barrio</dc:creator>
<dc:creator>Probir Chakravarty</dc:creator>
<dc:creator>Rebecca A Jones</dc:creator>
<dc:creator>S. Neda Mousavy Gharavy</dc:creator>
<dc:creator>Despina Stamataki</dc:creator>
<dc:creator>Julien Delile</dc:creator>
<dc:creator>James C Smith</dc:creator>
<dc:creator>James Briscoe</dc:creator>
<dc:date>2020-07-12</dc:date>
<dc:identifier>doi:10.1101/2020.07.12.198994</dc:identifier>
<dc:title><![CDATA[Ventricular, atrial and outflow tract heart progenitors arise from spatially and molecularly distinct regions of the primitive streak.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.13.200212v1?rss=1">
<title>
<![CDATA[
SPACE exploration of chromatin proteome to reveal associated RNA-binding proteins 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.13.200212v1?rss=1"
</link>
<description><![CDATA[
RNA-binding proteins (RBPs) play diverse roles in regulating co-transcriptional RNA-processing and chromatin functions, but our knowledge of the repertoire of chromatin-associated RBPs (caRBPs) and their interactions with chromatin remains limited. Here, we developed SPACE (Silica Particle Assisted Chromatin Enrichment) to isolate global and regional chromatin components with high specificity and sensitivity, and SPACEmap to identify the chromatin-contact regions in proteins. Applied to mouse embryonic stem cells, SPACE identified 1,459 chromatin-associated proteins, [~]48% of which are annotated as RBPs, indicating their dual roles in chromatin and RNA-binding. Additionally, SPACEmap stringently verified chromatin-binding of 404 RBPs and identified their chromatin-contact regions. Notably, SPACEmap showed that about half of the caRBPs bind to chromatin by intrinsically disordered regions (IDRs). Studying SPACE and total proteome dynamics from mES cells grown in 2iL and serum medium indicates significant correlation (R = 0.62). One of the most dynamic caRBPs is Dazl, which we find co-localized with PRC2 at transcription start sites of genes that are distinct from Dazl mRNA binding. Dazl and other PRC2-colocalised caRBPs are rich in intrinsically disordered regions (IDRs), which could contribute to the formation and regulation of phase-separated PRC condensates. Together, our approach provides an unprecedented insight into IDR-mediated interactions and caRBPs with moonlighting functions in native chromatin.
]]></description>
<dc:creator>Rafiee, M.-r.</dc:creator>
<dc:creator>Zagalak, J.</dc:creator>
<dc:creator>Tyzack, G.</dc:creator>
<dc:creator>Patani, R.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:date>2020-07-13</dc:date>
<dc:identifier>doi:10.1101/2020.07.13.200212</dc:identifier>
<dc:title><![CDATA[SPACE exploration of chromatin proteome to reveal associated RNA-binding proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.13.200444v1?rss=1">
<title>
<![CDATA[
Resolving phylogeny and polyploid parentage using genus-wide genome-wide sequence data from birch trees 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.13.200444v1?rss=1"
</link>
<description><![CDATA[
Numerous plant genera have a history including frequent hybridisation and polyploidisation, which often means that their phylogenies are not yet fully resolved. The genus Betula, which contains many ecologically important allopolyploid tree species, is a case in point. We generated genome-wide sequence data for 27 diploid and 31 polyploid Betula species or subspecies using restriction site associated DNA (RAD) sequences assembled into contigs with a mean length of 675 bp. We reconstructed the evolutionary relationships among diploid Betula species using both supermatrix and species tree methods. We identified progenitors of the polyploids according to the relative rates at which their reads mapped to contigs from different diploid species. We sorted the polyploid reads into different putative sub-genomes and used the extracted contigs, along with the diploid sequences, to build new phylogenies that included the polyploid sub-genomes. This approach yielded a highly evidenced phylogenetic hypothesis for the genus Betula, including the complex reticulate origins of the majority of its polyploid taxa. The genus was split into two well supported clades, which differ in their seed-wing morphology. We propose a new taxonomy for Betula, splitting it into two subgenera. We have resolved the parentage of many widespread and economically important polyploid tree species, opening the way for their population genomic study.
]]></description>
<dc:creator>Wang, N.</dc:creator>
<dc:creator>Kelly, L.</dc:creator>
<dc:creator>McAllister, H. A.</dc:creator>
<dc:creator>Zohren, J.</dc:creator>
<dc:creator>Buggs, R.</dc:creator>
<dc:date>2020-07-13</dc:date>
<dc:identifier>doi:10.1101/2020.07.13.200444</dc:identifier>
<dc:title><![CDATA[Resolving phylogeny and polyploid parentage using genus-wide genome-wide sequence data from birch trees]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.13.201459v1?rss=1">
<title>
<![CDATA[
Semi-Supervised Learning of Protein Secondary Structure from Single Sequences 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.13.201459v1?rss=1"
</link>
<description><![CDATA[
Accurate modelling of a single orphan protein sequence in the absence of homology information has remained a challenge for several decades. Although not as performant as their homology-based counterparts, single-sequence bioinformatic methods are not constrained by the requirement of evolutionary information and so have a swathe of applications and uses. By taking a bioinformatics approach to semi-supervised machine learning we develop Profile Augmentation of Single Sequences (PASS), a simple but powerful framework for developing accurate single-sequence methods. To demonstrate the effectiveness of PASS we apply it to the mature field of secondary structure prediction. In doing so we develop S4PRED, the successor to the open-source PSIPRED-Single method, which achieves an unprecedented Q3 score of 75.3% on the standard CB513 test. PASS provides a blueprint for the development of a new generation of predictive methods, advancing our ability to model individual protein sequences.
]]></description>
<dc:creator>Moffat, L.</dc:creator>
<dc:creator>Jones, D. T.</dc:creator>
<dc:date>2020-07-14</dc:date>
<dc:identifier>doi:10.1101/2020.07.13.201459</dc:identifier>
<dc:title><![CDATA[Semi-Supervised Learning of Protein Secondary Structure from Single Sequences]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.15.204107v1?rss=1">
<title>
<![CDATA[
The Mycobacterium tuberculosis sRNA F6 regulates expression of groEL/S 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.15.204107v1?rss=1"
</link>
<description><![CDATA[
Almost 140 years after the identification of Mycobacterium tuberculosis as the etiological agent of tuberculosis, important aspects of its biology remain poorly described. Little is known about the role of post-transcriptional control of gene expression and RNA biology, including the role of most of the small RNAs (sRNAs) identified to date. We have carried out a detailed investigation of the M. tuberculosis sRNA, F6, and shown it to be dependent on SigF for expression, and significantly induced in starvation conditions in vitro and in a mouse model of infection. Further exploration of F6 using an in vitro starvation model of infection indicates that F6 affects the expression of the essential chaperonins, GroEL2 and GroES. Our results point towards a role for F6 during periods of low metabolic activity typically associated with long-term survival of M. tuberculosis in human granulomas.
]]></description>
<dc:creator>Houghton, J.</dc:creator>
<dc:creator>Rodgers, A.</dc:creator>
<dc:creator>Rose, G.</dc:creator>
<dc:creator>Arnvig, K. B.</dc:creator>
<dc:date>2020-07-15</dc:date>
<dc:identifier>doi:10.1101/2020.07.15.204107</dc:identifier>
<dc:title><![CDATA[The Mycobacterium tuberculosis sRNA F6 regulates expression of groEL/S]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.16.199083v1?rss=1">
<title>
<![CDATA[
Quantifying the monomer-dimer equilibrium of tubulin with mass photometry 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.16.199083v1?rss=1"
</link>
<description><![CDATA[
The {beta}-tubulin heterodimer is the fundamental building block of microtubules, making it central to several cellular processes. Despite the apparent simplicity of heterodimerisation, the associated energetics and kinetics remain disputed, largely due to experimental challenges associated with quantifying affinities in the <M range. We use mass photometry to observe tubulin monomers and heterodimers in solution simultaneously, thereby quantifying the {beta}-tubulin dissociation constant (8.48{+/-}1.22 nM) and its tightening in the presence of GTP (3.69{+/-}0.65 nM), at a dissociation rate >10-2 s-1. Our results demonstrate the capabilities of mass photometry for quantifying protein-protein interactions and clarify the energetics and kinetics of tubulin heterodimerisation.



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]]></description>
<dc:creator>Fineberg, A.</dc:creator>
<dc:creator>Surrey, T.</dc:creator>
<dc:creator>Kukura, P.</dc:creator>
<dc:date>2020-07-16</dc:date>
<dc:identifier>doi:10.1101/2020.07.16.199083</dc:identifier>
<dc:title><![CDATA[Quantifying the monomer-dimer equilibrium of tubulin with mass photometry]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.15.205732v1?rss=1">
<title>
<![CDATA[
The malaria parasite sheddase SUB2 governs host red blood cell membrane sealing at invasion 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.15.205732v1?rss=1"
</link>
<description><![CDATA[
Red blood cell (RBC) invasion by malaria merozoites involves formation of a parasitophorous vacuole into which the parasite moves. The vacuole membrane seals and pinches off behind the parasite through an unknown mechanism, enclosing the parasite within the RBC. During invasion, several parasite surface proteins are shed by a membrane-bound protease called SUB2. Here we show that genetic depletion of SUB2 abolishes shedding of a range of parasite proteins, identifying previously unrecognized SUB2 substrates. Interaction of SUB2-null merozoites with RBCs leads to either abortive invasion with rapid RBC lysis, or successful entry but developmental arrest. Selective failure to shed the most abundant SUB2 substrate, MSP1, reduces intracellular replication, whilst conditional ablation of the substrate AMA1 produces host RBC lysis. We conclude that SUB2 activity is critical for host RBC membrane sealing following parasite internalisation and for correct functioning of merozoite surface proteins.

Key highlightsO_LIMany malaria parasite surface proteins are shed by SUB2 during RBC invasion
C_LIO_LISUB2-null merozoites either induce rapid host RBC lysis, or invade then die
C_LIO_LIMerozoite surface protein shedding is crucial for host RBC membrane sealing
C_LI
]]></description>
<dc:creator>Collins, C. R.</dc:creator>
<dc:creator>Hackett, F.</dc:creator>
<dc:creator>Howell, S. A.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Russell, M. R.</dc:creator>
<dc:creator>Collinson, L. M.</dc:creator>
<dc:creator>Blackman, M. J.</dc:creator>
<dc:date>2020-07-17</dc:date>
<dc:identifier>doi:10.1101/2020.07.15.205732</dc:identifier>
<dc:title><![CDATA[The malaria parasite sheddase SUB2 governs host red blood cell membrane sealing at invasion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.18.209973v1?rss=1">
<title>
<![CDATA[
A multipass membrane protein interacts with the cGMP-dependent protein kinase to regulate critical calcium signals in malaria parasites 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.18.209973v1?rss=1"
</link>
<description><![CDATA[
In malaria parasites, all cGMP-dependent signalling is mediated through a single cGMP-dependent protein kinase (PKG), a major function of which is to control essential calcium signals. However, how PKG transmits these signals in the absence of known second messenger-dependent calcium channels or scaffolding proteins is unknown. Here we identify a polytopic membrane protein, ICM1, with homology to transporters and calcium channels that is tightly-associated with PKG in both Plasmodium falciparum asexual blood stages and P. berghei gametocytes. Phosphoproteomic analyses in both Plasmodium species reveal multiple ICM1 phosphorylation events dependent upon PKG activity. Stage-specific depletion of P. berghei ICM1 blocks gametogenesis due to the inability of mutant parasites to mobilise intracellular calcium upon PKG activation, whilst conditional loss of P. falciparum ICM1 results in reduced calcium mobilisation, defective egress and lack of invasion. Our findings provide new insights into atypical calcium homeostasis in malaria parasites essential for pathology and disease transmission.
]]></description>
<dc:creator>Balestra, A. C.</dc:creator>
<dc:creator>Koussis, K.</dc:creator>
<dc:creator>Klages, N.</dc:creator>
<dc:creator>Howell, S. A.</dc:creator>
<dc:creator>Flynn, H. R.</dc:creator>
<dc:creator>Bantscheff, M.</dc:creator>
<dc:creator>Pasquarello, C.</dc:creator>
<dc:creator>Perrin, A. J.</dc:creator>
<dc:creator>Brusini, L.</dc:creator>
<dc:creator>Arboit, P.</dc:creator>
<dc:creator>Sanz, O.</dc:creator>
<dc:creator>Peces-Barba Castano, L.</dc:creator>
<dc:creator>Withers-Martinez, C.</dc:creator>
<dc:creator>Hainard, A.</dc:creator>
<dc:creator>Ghidelli-Disse, S.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Baker, D. A.</dc:creator>
<dc:creator>Blackman, M. J.</dc:creator>
<dc:creator>Brochet, M.</dc:creator>
<dc:date>2020-07-18</dc:date>
<dc:identifier>doi:10.1101/2020.07.18.209973</dc:identifier>
<dc:title><![CDATA[A multipass membrane protein interacts with the cGMP-dependent protein kinase to regulate critical calcium signals in malaria parasites]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.20.211557v1?rss=1">
<title>
<![CDATA[
An aberrant cytoplasmic intron retention programme is a blueprint for ALS-related RBP mislocalization 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.20.211557v1?rss=1"
</link>
<description><![CDATA[
We recently described aberrant cytoplasmic SFPQ intron-retaining transcripts (IRTs) and concurrent SFPQ protein mislocalization as a new hallmark of amyotrophic lateral sclerosis (ALS). However the generalizability and potential roles of cytoplasmic IRTs in health and disease remain unclear. Here, using time-resolved deep-sequencing of nuclear and cytoplasmic fractions of hiPSCs undergoing motor neurogenesis, we reveal that ALS-causing VCP gene mutations lead to compartment-specific aberrant accumulation of IRTs. Specifically, we identify >100 IRTs with increased cytoplasmic (but not nuclear) abundance in ALS samples. Furthermore, these aberrant cytoplasmic IRTs possess sequence-specific attributes and differential predicted binding affinity to RNA binding proteins (RBPs). Remarkably, TDP-43, SFPQ and FUS - RBPs known for nuclear-to-cytoplasmic mislocalization in ALS - avidly and specifically bind to this aberrant cytoplasmic pool of IRTs, as opposed to any individual IRT. Our data are therefore consistent with a novel role for cytoplasmic IRTs in regulating compartment-specific protein abundance. This study provides new molecular insight into potential pathomechanisms underlying ALS and highlights aberrant cytoplasmic IRTs as potential therapeutic targets.



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]]></description>
<dc:creator>Tyzack, G. E.</dc:creator>
<dc:creator>Neeves, J.</dc:creator>
<dc:creator>Klein, P.</dc:creator>
<dc:creator>Crerar, H.</dc:creator>
<dc:creator>Ziff, O.</dc:creator>
<dc:creator>Taha, D. M.</dc:creator>
<dc:creator>Luisier, R.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:creator>Patani, R.</dc:creator>
<dc:date>2020-07-21</dc:date>
<dc:identifier>doi:10.1101/2020.07.20.211557</dc:identifier>
<dc:title><![CDATA[An aberrant cytoplasmic intron retention programme is a blueprint for ALS-related RBP mislocalization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.20.208835v1?rss=1">
<title>
<![CDATA[
HIV coinfection is associated with low fitness rpoB variants in rifampicin-resistant Mycobacterium tuberculosis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.20.208835v1?rss=1"
</link>
<description><![CDATA[
We analysed 312 drug-resistant genomes of Mycobacterium tuberculosis (Mtb) collected from HIV coinfected and HIV negative TB patients from nine countries with a high tuberculosis burden. We found that rifampicin-resistant Mtb strains isolated from HIV coinfected patients carried disproportionally more resistance-conferring mutations in rpoB that are associated with a low fitness in the absence of the drug, suggesting these low fitness rpoB variants can thrive in the context of reduced host immunity.
]]></description>
<dc:creator>Loiseau, C.</dc:creator>
<dc:creator>Brites, D.</dc:creator>
<dc:creator>Reinhard, M.</dc:creator>
<dc:creator>Zuercher, K.</dc:creator>
<dc:creator>Borrell, S.</dc:creator>
<dc:creator>Ballif, M.</dc:creator>
<dc:creator>Fenner, L.</dc:creator>
<dc:creator>Cox, H.</dc:creator>
<dc:creator>Rutaihwa, L. K.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Yotebieng, M.</dc:creator>
<dc:creator>Carter, E. J.</dc:creator>
<dc:creator>Abimiku, A.</dc:creator>
<dc:creator>Marcy, O.</dc:creator>
<dc:creator>Gotuzzo, E.</dc:creator>
<dc:creator>Avihingsanon, A.</dc:creator>
<dc:creator>Zetola, N.</dc:creator>
<dc:creator>Doulla, B.</dc:creator>
<dc:creator>Boettger, E. C.</dc:creator>
<dc:creator>Egger, M.</dc:creator>
<dc:creator>Gagneux, S.</dc:creator>
<dc:date>2020-07-22</dc:date>
<dc:identifier>doi:10.1101/2020.07.20.208835</dc:identifier>
<dc:title><![CDATA[HIV coinfection is associated with low fitness rpoB variants in rifampicin-resistant Mycobacterium tuberculosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.21.213470v1?rss=1">
<title>
<![CDATA[
Protective porcine influenza virus-specific monoclonal antibodies recognize similar haemagglutinin epitopes as humans 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.21.213470v1?rss=1"
</link>
<description><![CDATA[
Pigs are natural hosts for the same subtypes of influenza A viruses as humans and integrally involved in virus evolution with frequent interspecies transmissions in both directions. The emergence of the 2009 pandemic H1N1 virus illustrates the importance of pigs in evolution of zoonotic strains. Here we generated pig influenza-specific monoclonal antibodies (mAbs) from H1N1pdm09 infected pigs. The mAbs recognized the same two major immunodominant haemagglutinin (HA) epitopes targeted by humans, one of which is not recognized by post-infection ferret antisera that are commonly used to monitor virus evolution. Neutralizing activity of the pig mAbs was comparable to that of potent human anti-HA mAbs. Further, prophylactic administration of a selected porcine mAb to pigs abolished lung viral load and greatly reduced lung pathology but did not eliminate nasal shedding of virus after H1N1pdm09 challenge. Hence mAbs from pigs, which target HA can significantly reduce disease severity. These results, together with the comparable sizes of pigs and humans, indicate that the pig is a valuable model for understanding how best to apply mAbs as therapy in humans and for monitoring antigenic drift of influenza viruses in humans, thereby providing information highly relevant to making influenza vaccine recommendations.
]]></description>
<dc:creator>Holzer, B.</dc:creator>
<dc:creator>Rijal, P.</dc:creator>
<dc:creator>McNee, A.</dc:creator>
<dc:creator>Paudyal, B.</dc:creator>
<dc:creator>Clark, B.</dc:creator>
<dc:creator>Manjegowda, T.</dc:creator>
<dc:creator>Salguero, F. J.</dc:creator>
<dc:creator>Bessell, E.</dc:creator>
<dc:creator>Schwartz, J. C.</dc:creator>
<dc:creator>Moffat, K.</dc:creator>
<dc:creator>Pedrera, M.</dc:creator>
<dc:creator>Graham, S. P.</dc:creator>
<dc:creator>Placido, M. B.-D.</dc:creator>
<dc:creator>La Ragione, R. M.</dc:creator>
<dc:creator>Mwangi, W.</dc:creator>
<dc:creator>Beverley, P.</dc:creator>
<dc:creator>McCauley, J. W.</dc:creator>
<dc:creator>Daniels, R. S.</dc:creator>
<dc:creator>Hammond, J.</dc:creator>
<dc:creator>Townsend, A. R.</dc:creator>
<dc:creator>Tchilian, E.</dc:creator>
<dc:date>2020-07-22</dc:date>
<dc:identifier>doi:10.1101/2020.07.21.213470</dc:identifier>
<dc:title><![CDATA[Protective porcine influenza virus-specific monoclonal antibodies recognize similar haemagglutinin epitopes as humans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.24.214551v1?rss=1">
<title>
<![CDATA[
Phase Unwrapping with a Rapid Opensource Minimum Spanning TreE AlgOrithm (ROMEO) 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.24.214551v1?rss=1"
</link>
<description><![CDATA[
PurposeTo develop a rapid and accurate MRI phase unwrapping technique for challenging phase topographies encountered at high magnetic fields, around metal implants or post-operative cavities, that is sufficiently fast to be applied to large group studies including Quantitative Susceptibility Mapping and functional MRI (with phase-based distortion correction).

MethodsThe proposed path-following phase unwrapping algorithm, ROMEO, estimates the coherence of the signal both in space - using MRI magnitude and phase information - and over time, assuming approximately linear temporal phase evolution. This information is combined to form a quality map that guides the unwrapping along a three-dimensional path through the object using a computationally efficient minimum spanning tree algorithm. ROMEO was tested against the two most commonly used exact phase unwrapping methods: PRELUDE and BEST PATH in simulated topographies and at several field strengths: in 3 T and 7 T in vivo human head images and 9.4 T ex vivo rat head images.

ResultsROMEO was more reliable than PRELUDE and BEST PATH, yielding unwrapping results with excellent temporal stability for multi-echo or multi-time-point data. ROMEO does not require image masking and delivers results within seconds even in large, highly wrapped multi-echo datasets (e.g. 9 seconds for a 7 T head dataset with 31 echoes and a 208 x 208 x 96 matrix size).

ConclusionOverall, ROMEO was both faster and more accurate than PRELUDE and BEST PATH delivering exact results within seconds, which is well below typical image acquisition times, enabling potential on-console application.
]]></description>
<dc:creator>Dymerska, B.</dc:creator>
<dc:creator>Eckstein, K.</dc:creator>
<dc:creator>Bachrata, B.</dc:creator>
<dc:creator>Siow, B.</dc:creator>
<dc:creator>Trattnig, S.</dc:creator>
<dc:creator>Shmueli, K.</dc:creator>
<dc:creator>Robinson, S. D.</dc:creator>
<dc:date>2020-07-24</dc:date>
<dc:identifier>doi:10.1101/2020.07.24.214551</dc:identifier>
<dc:title><![CDATA[Phase Unwrapping with a Rapid Opensource Minimum Spanning TreE AlgOrithm (ROMEO)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.24.219139v1?rss=1">
<title>
<![CDATA[
Tissue-specific and interferon-inducible expression of non-functional ACE2 through endogenous retrovirus co-option 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.24.219139v1?rss=1"
</link>
<description><![CDATA[
Angiotensin-converting enzyme 2 (ACE2) is an entry receptor for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), as well as a regulator of several physiological processes. ACE2 has recently been proposed to be interferon-inducible, suggesting that SARS-CoV-2 may exploit this phenomenon to enhance viral spread and questioning the efficacy of interferon treatment in Coronavirus disease 2019 (COVID-19). Using a recent de novo transcript assembly that captured previously unannotated transcripts, we describe a novel isoform of ACE2, generated by co-option of an intronic long terminal repeat (LTR) retroelement promoter. The novel transcript, termed LTR16A1-ACE2, exhibits specific expression patterns across the aerodigestive and gastrointestinal tracts and, importantly, is highly responsive to interferon stimulation. In stark contrast, expression of canonical ACE2 is completely unresponsive to interferon stimulation. Moreover, the LTR16A1-ACE2 translation product is a truncated, unstable ACE2 form, lacking domains required for SARS-CoV-2 binding and therefore unlikely to contribute to or enhance viral infection.
]]></description>
<dc:creator>Ng, K.</dc:creator>
<dc:creator>Attig, J.</dc:creator>
<dc:creator>Bolland, W.</dc:creator>
<dc:creator>Young, G.</dc:creator>
<dc:creator>Major, J.</dc:creator>
<dc:creator>Wack, A.</dc:creator>
<dc:creator>Kassiotis, G.</dc:creator>
<dc:date>2020-07-24</dc:date>
<dc:identifier>doi:10.1101/2020.07.24.219139</dc:identifier>
<dc:title><![CDATA[Tissue-specific and interferon-inducible expression of non-functional ACE2 through endogenous retrovirus co-option]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.28.223024v1?rss=1">
<title>
<![CDATA[
Citizen science, cells and CNNs - deep learning for automatic segmentation of the nuclear envelope in electron microscopy data, trained with volunteer segmentations 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.28.223024v1?rss=1"
</link>
<description><![CDATA[
Advancements in volume electron microscopy mean it is now possible to generate thousands of serial images at nanometre resolution overnight, yet the gold standard approach for data analysis remains manual segmentation by an expert microscopist, resulting in a critical research bottleneck. Although some machine learning approaches exist in this domain, we remain far from realising the aspiration of a highly accurate, yet generic, automated analysis approach, with a major obstacle being lack of sufficient high-quality ground-truth data. To address this, we developed a novel citizen science project, Etch a Cell, to enable volunteers to manually segment the nuclear envelope of HeLa cells imaged with Serial Blockface SEM. We present our approach for aggregating multiple volunteer annotations to generate a high quality consensus segmentation, and demonstrate that data produced exclusively by volunteers can be used to train a highly accurate machine learning algorithm for automatic segmentation of the nuclear envelope, which we share here, in addition to our archived benchmark data.
]]></description>
<dc:creator>Spiers, H.</dc:creator>
<dc:creator>Songhurst, H.</dc:creator>
<dc:creator>Nightingale, L.</dc:creator>
<dc:creator>de Folter, J.</dc:creator>
<dc:creator>Hutchings, R.</dc:creator>
<dc:creator>Peddie, C. J.</dc:creator>
<dc:creator>Weston, A.</dc:creator>
<dc:creator>Strange, A.</dc:creator>
<dc:creator>Hindmarsh, S.</dc:creator>
<dc:creator>Lintott, C.</dc:creator>
<dc:creator>Collinson, L. M.</dc:creator>
<dc:creator>Jones, M. L.</dc:creator>
<dc:date>2020-07-29</dc:date>
<dc:identifier>doi:10.1101/2020.07.28.223024</dc:identifier>
<dc:title><![CDATA[Citizen science, cells and CNNs - deep learning for automatic segmentation of the nuclear envelope in electron microscopy data, trained with volunteer segmentations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.29.226357v1?rss=1">
<title>
<![CDATA[
Control and mechanisms of pulsatile flows in epithelial monolayers 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.29.226357v1?rss=1"
</link>
<description><![CDATA[
Collective motions of epithelial cells in vivo are essential for morphogenesis in developmental biology. Tissues elongate, contract, flow, and oscillate, thus sculpting embryos. These tissue level dynamics are known, but the physical mechanisms at the cellular level are unclear, with various behaviors depending on the tissues and species. Moreover, investigations on in vitro tissue behavior usually focus on only one type of cell dynamics and use diverse theoretical approaches, making systematic comparisons between studies challenging. Here, we show that a single epithelial monolayer of Madin Darby Canine Kidney (MDCK) cells can exhibit two types of local tissue kinematics, pulsations and long range coherent flows. We analyzed these distinct motions by using quantitative live imaging. We also report that these motions can be controlled with internal and external cues such as specific inhibitors, and friction modulation of the substrate by microcontact printing method. We further demonstrate with a unified vertex model that both behaviors depend on the competition between velocity alignment and random diffusion of cell polarization. When alignment and diffusion are comparable, a pulsatile flow emerges, whereas the tissue undergoes long-range flows when velocity alignment dominates. We propose that environmental friction, acto-myosin distributions, and cell polarization kinetics are important in regulating the dynamics of tissue morphogenesis.
]]></description>
<dc:creator>Thiagarajan, R.</dc:creator>
<dc:creator>Bhat, A.</dc:creator>
<dc:creator>Salbreux, G.</dc:creator>
<dc:creator>Inamdar, M. M.</dc:creator>
<dc:creator>Riveline, D.</dc:creator>
<dc:date>2020-07-30</dc:date>
<dc:identifier>doi:10.1101/2020.07.29.226357</dc:identifier>
<dc:title><![CDATA[Control and mechanisms of pulsatile flows in epithelial monolayers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.08.03.230615v1?rss=1">
<title>
<![CDATA[
Maternal iron deficiency perturbs embryonic cardiovascular development 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.03.230615v1?rss=1"
</link>
<description><![CDATA[
Congenital heart disease (CHD) is the most common type of birth defect, with a global prevalence of 0.9% of live births1. Most research in the last 30 years has focused on finding genetic causes of CHD. However, despite the association of over 100 genes with CHD, mutations in these genes only explain ~30% of cases2. Many of the remaining cases of CHD are caused by in utero exposure to environmental factors3. Here we have identified a completely new environmental teratogen causing CHD: maternal iron deficiency. In humans, iron deficiency anaemia is a major global health problem. 38% of pregnant women worldwide are anaemic4, and at least half of these are due to iron deficiency, the most prevalent micronutrient deficiency. We describe a mouse model of maternal iron deficiency anaemia that causes severe cardiovascular defects in her offspring. We show that these defects likely arise from increased retinoic acid signalling in iron deficient embryos, probably due to reduced activity of the iron-dependent retinoic acid catabolic CYP26 enzymes. The defects can be prevented by maternal iron administration early in pregnancy, and are also greatly reduced in offspring of mothers deficient in both iron and the retinoic acid precursor vitamin A. Finally, one puzzling feature of many genetic forms of CHD in humans is the considerable variation in penetrance and severity of defects. We show that maternal iron deficiency acts as a significant modifier of heart and craniofacial phenotype in a mouse model of Down syndrome. Given the high incidence of maternal iron deficiency, peri-conceptional iron monitoring and supplementation could be a viable strategy to reduce the prevalence and severity of CHD in human populations worldwide.
]]></description>
<dc:creator>Kalisch-Smith, J. I.</dc:creator>
<dc:creator>Ved, N.</dc:creator>
<dc:creator>Szumska, D.</dc:creator>
<dc:creator>Munro, J.</dc:creator>
<dc:creator>Troup, M.</dc:creator>
<dc:creator>Harris, S. E.</dc:creator>
<dc:creator>Jacquemot, A.</dc:creator>
<dc:creator>Miller, J. J.</dc:creator>
<dc:creator>Stuart, E. M.</dc:creator>
<dc:creator>Wolna, M.</dc:creator>
<dc:creator>Hardman, E.</dc:creator>
<dc:creator>Prin, F.</dc:creator>
<dc:creator>Lana-Elola, E.</dc:creator>
<dc:creator>Aoidi, R.</dc:creator>
<dc:creator>Fisher, E. M. C.</dc:creator>
<dc:creator>Tybulewicz, V. L. J.</dc:creator>
<dc:creator>Mohun, T. J.</dc:creator>
<dc:creator>Lakhal-Littleton, S.</dc:creator>
<dc:creator>Giannoulatou, E.</dc:creator>
<dc:creator>Sparrow, D. B.</dc:creator>
<dc:date>2020-08-03</dc:date>
<dc:identifier>doi:10.1101/2020.08.03.230615</dc:identifier>
<dc:title><![CDATA[Maternal iron deficiency perturbs embryonic cardiovascular development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.08.05.237537v1?rss=1">
<title>
<![CDATA[
Correlative Light Electron Ion Microscopy reveal in vivo localisation of bedaquiline in Mycobacterium tuberculosis infected lungs 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.05.237537v1?rss=1"
</link>
<description><![CDATA[
Correlative light, electron and ion microscopy (CLEIM) offers huge potential to track the intracellular fate of antibiotics, with organelle-level resolution. However, a correlative approach that enables subcellular antibiotic visualisation in pathogen-infected tissue is lacking. Here, we developed CLEIM in tissue (CLEIMiT), and used it to identify the cell-type specific accumulation of an antibiotic in lung lesions of mice infected with Mycobacterium tuberculosis. Using CLEIMiT, we found that the anti-TB drug bedaquiline is localised not only in foamy macrophages in the lungs during infection but also accumulate in polymorphonuclear (PMN) cells.
]]></description>
<dc:creator>Fearns, A.</dc:creator>
<dc:creator>Greenwood, D.</dc:creator>
<dc:creator>Rodgers, A.</dc:creator>
<dc:creator>Jiang, H.</dc:creator>
<dc:creator>Gutierrez, M.</dc:creator>
<dc:date>2020-08-05</dc:date>
<dc:identifier>doi:10.1101/2020.08.05.237537</dc:identifier>
<dc:title><![CDATA[Correlative Light Electron Ion Microscopy reveal in vivo localisation of bedaquiline in Mycobacterium tuberculosis infected lungs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.08.07.241042v1?rss=1">
<title>
<![CDATA[
Theory of genetic oscillations with external noisy regulation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.07.241042v1?rss=1"
</link>
<description><![CDATA[
We present a general theory of noisy genetic oscillators with externally regulated production rate. The observables that characterize the genetic oscillator are discussed, and it is shown how their statistics depend on the statistics of the external regulator. We show that these observables have generic features that are observed in two different experimental systems: the expression of the circadian clock genes in fibroblasts, and in the transient and oscillatory dynamics of the segmentation clock genes observed in cells disassociated from zebrafish embryos. Our work shows that genetic oscillations with diverse biological contexts can be understood in a common framework based on delayed negative feedback system, and slow regulator dynamics.
]]></description>
<dc:creator>Negrete, J.</dc:creator>
<dc:creator>Lengyel, I. M.</dc:creator>
<dc:creator>Rohde, L.</dc:creator>
<dc:creator>Desai, R. A.</dc:creator>
<dc:creator>Oates, A. C.</dc:creator>
<dc:creator>Julicher, F.</dc:creator>
<dc:date>2020-08-07</dc:date>
<dc:identifier>doi:10.1101/2020.08.07.241042</dc:identifier>
<dc:title><![CDATA[Theory of genetic oscillations with external noisy regulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.08.06.239822v1?rss=1">
<title>
<![CDATA[
Cellular barcoding of Toxoplasma reveals permissive host brain colonization 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.06.239822v1?rss=1"
</link>
<description><![CDATA[
Molecular barcoding techniques have emerged as powerful tools to understand microbial pathogenesis. However, barcoding strategies have not been extended to protozoan parasites, which have unique genomic structures and virulence strategies compared to viral and bacterial pathogens. Here, we present a versatile CRISPR-based method to barcode protozoa, which we successfully apply to Toxoplasma gondii and Trypanosoma brucei. The murine brain is an important transmission niche for T. gondii, and brain persistence is a clinically untreatable feature of infection. The blood-brain barrier is expected to physically restrict parasite colonization of this niche, resulting in a selection bottleneck. Using libraries of barcoded T. gondii we evaluate shifts in the population structure from acute to chronic infection of mice. Contrary to expectation, most barcodes were present in the brain one-month post-intraperitoneal infection in both inbred CBA/J and outbred Swiss mice. Although parasite cyst number and barcode diversity declined over time, barcodes that represented a minor fraction of the inoculum could become a dominant population in the brain by three months post-infection. Together, these data establish the first, robust molecular barcoding approach for protozoa and evidence that the blood-brain barrier does not represent a major bottleneck to colonization by T. gondii.
]]></description>
<dc:creator>Wincott, C. J.</dc:creator>
<dc:creator>Sritharan, G.</dc:creator>
<dc:creator>Bunyan, M.</dc:creator>
<dc:creator>Alves, E.</dc:creator>
<dc:creator>Benns, H. J.</dc:creator>
<dc:creator>Frickel, E. M.</dc:creator>
<dc:creator>Ewald, S. E.</dc:creator>
<dc:creator>Child, M. A.</dc:creator>
<dc:date>2020-08-07</dc:date>
<dc:identifier>doi:10.1101/2020.08.06.239822</dc:identifier>
<dc:title><![CDATA[Cellular barcoding of Toxoplasma reveals permissive host brain colonization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.08.11.245860v1?rss=1">
<title>
<![CDATA[
Single amino acid-promoted reactions link a non-enzymatic chemical network to the early evolution of enzymatic pentose phosphate pathway 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.11.245860v1?rss=1"
</link>
<description><![CDATA[
How metabolic pathways emerged in early evolution remains largely unknown. Recently discovered chemical networks driven by iron and sulfur resemble reaction sequences found within glycolysis, gluconeogenesis, the oxidative and reductive Krebs cycle, the Wood Ljungdahl as well as the S-adenosylmethionine pathways, components of the core cellular metabolic network. These findings suggest that the evolution of central metabolism was primed by environmental chemical reactions, implying that non-enzymatic reaction networks served as a "template" in the evolution of enzymatic activities. We speculated that the turning point for this transition would depend on the catalytic properties of the simplest structural components of proteins, single amino acids. Here, we systematically combine constituents of Fe(II)-driven non-enzymatic reactions resembling glycolysis and pentose phosphate pathway (PPP), with single proteinogenic amino acids. Multiple reaction rates are enhanced by amino acids. In particular, cysteine is able to replace (and/or complement) the metal ion Fe(II) in driving the non-enzymatic formation of the RNA-backbone metabolite ribose 5-phosphate from 6-phosphogluconate, a rate-limiting reaction of the oxidative PPP. In the presence of both Fe(II) and cysteine, a complex is formed, enabling the non-enzymatic reaction to proceed at a wide range of temperatures. At mundane temperatures, this  minimal enzyme-like complex achieves a much higher specificity in the formation of ribose 5-phosphate than the Fe(II)-driven reaction at high temperatures. Hence, simple amino acids can accelerate key steps within metal-promoted metabolism-like chemical networks. Our results imply a stepwise scenario, in which environmental chemical networks served as primers in the early evolution of the metabolic network structure.

Significance StatementThe evolutionary roots of metabolic pathways are barely understood. Here we show results consistent with a stepwise scenario during the evolution of (enzymatic) metabolism, starting from non-enzymatic chemical networks. By systematic screening of metabolic-like reactivities in vitro, and using high-throughput analytical techniques, we identify an iron/cysteine complex to act as a  minimal enzymelike complex, which consists of a metal ion, an amino acid, and a sugar phosphate ligand. Integrated in a metal-driven, non-enzymatic pentose phosphate pathway, it promotes the formation of the RNA-backbone precursor ribose 5-phosphate at ambient temperature.
]]></description>
<dc:creator>Piedrafita, G.</dc:creator>
<dc:creator>Varma, S. J.</dc:creator>
<dc:creator>Castro, C.</dc:creator>
<dc:creator>Messner, C. B.</dc:creator>
<dc:creator>Szyrwiel, L.</dc:creator>
<dc:creator>Griffin, J. L.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:date>2020-08-12</dc:date>
<dc:identifier>doi:10.1101/2020.08.11.245860</dc:identifier>
<dc:title><![CDATA[Single amino acid-promoted reactions link a non-enzymatic chemical network to the early evolution of enzymatic pentose phosphate pathway]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.08.11.245621v1?rss=1">
<title>
<![CDATA[
PairGP: Gaussian process modeling of longitudinal data from paired multi-condition studies 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.11.245621v1?rss=1"
</link>
<description><![CDATA[
We propose PairGP, a non-stationary Gaussian process method to compare gene expression timeseries across several conditions that can account for paired longitudinal study designs and can identify groups of conditions that have different gene expression dynamics. We demonstrate the method on both simulated data and previously unpublished RNA-seq time-series with five conditions. The results show the advantage of modeling the pairing effect to better identify groups of conditions with different dynamics. The implementations is available at https://github.com/michelevantini/PairGP
]]></description>
<dc:creator>Vantini, M.</dc:creator>
<dc:creator>Mannerström, H.</dc:creator>
<dc:creator>Rautio, S.</dc:creator>
<dc:creator>Ahlfors, H.</dc:creator>
<dc:creator>Stockinger, B.</dc:creator>
<dc:creator>Lähdesmäki, H.</dc:creator>
<dc:date>2020-08-12</dc:date>
<dc:identifier>doi:10.1101/2020.08.11.245621</dc:identifier>
<dc:title><![CDATA[PairGP: Gaussian process modeling of longitudinal data from paired multi-condition studies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.08.11.245969v1?rss=1">
<title>
<![CDATA[
Cisplatin is more mutagenic than carboplatin or oxaliplatin at equitoxic concentrations 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.11.245969v1?rss=1"
</link>
<description><![CDATA[
Platinum-based drugs are a mainstay of cancer chemotherapy. However, their mutagenic effect can increase tumour heterogeneity, contribute to the evolution of treatment resistance, and also induce secondary malignancies. We coupled whole genome sequencing with phenotypic investigations on two cell line models to compare the magnitude and understand the mechanism of mutagenicity of cisplatin, carboplatin and oxaliplatin. Cisplatin induced significantly more base substitution mutations than carboplatin or oxaliplatin when used at equitoxic concentrations on human TK6 or chicken DT40 cells, and also induced the highest number of short insertions and deletions. Assessment through histone H2AX phosphorylation and single cell agarose gel electrophoresis suggested that cisplatin caused more DNA damage than carboplatin or oxaliplatin. The analysis of base substitution spectra revealed that all three tested platinum drugs elicit both a direct mutagenic effect at purine dinucleotides, and an indirect effect of accelerating endogenous mutagenic processes. Whereas the direct mutagenic effect correlated with the level of DNA damage caused, the indirect mutagenic effects were equal. The different mutagenicity and DNA damaging effect of equitoxic platinum drug treatments suggests that DNA damage independent mechanisms significantly contribute to their cytotoxicity. Thus, the comparatively high mutagenicity of cisplatin should be taken into account in the design of chemotherapeutic regimens.
]]></description>
<dc:creator>Szikriszt, B.</dc:creator>
<dc:creator>Poti, A.</dc:creator>
<dc:creator>Nemeth, E.</dc:creator>
<dc:creator>Kanu, N.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Szuts, D.</dc:creator>
<dc:date>2020-08-12</dc:date>
<dc:identifier>doi:10.1101/2020.08.11.245969</dc:identifier>
<dc:title><![CDATA[Cisplatin is more mutagenic than carboplatin or oxaliplatin at equitoxic concentrations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.08.17.253773v1?rss=1">
<title>
<![CDATA[
Automated and unbiased classification of motor neuron phenotypes with single cell resolution in ALS tissue. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.17.253773v1?rss=1"
</link>
<description><![CDATA[
Histopathological analysis of tissue sections is an invaluable resource in neurodegeneration research. Importantly, cell-to-cell variation in both the presence and severity of a given phenotype is however a key limitation of this approach, reducing the signal to noise ratio and leaving unresolved the potential of single-cell scoring for a given disease attribute. Here, we developed an image processing pipeline for automated identification and profiling of motor neurons (MNs) in amyotrophic lateral sclerosis (ALS) pathological tissue sections. This approach enabled unbiased analysis of hundreds of cells, from which hundreds of features were readily extracted. Next by testing different machine learning methods, we automated the identification of phenotypically distinct MN subpopulations in VCP- and SOD1-mutant transgenic mice, revealing common aberrant phenotypes in cellular shape. Additionally we established scoring metrics to rank cells and tissue samples for both disease probability and severity. Finally, by adapting this methodology to human post-mortem tissue analysis, we validated our core finding that morphological descriptors strongly discriminate ALS from control healthy tissue at the single cell level. In summary, we show that combining automated image processing with machine learning methods substantially improves the speed and reliability of identifying phenotypically diverse MN populations. Determining disease presence, severity and unbiased phenotypes at single cell resolution might prove transformational in our understanding of ALS and neurodegenerative diseases more broadly.
]]></description>
<dc:creator>Luisier, R.</dc:creator>
<dc:creator>Serio, A.</dc:creator>
<dc:creator>Patani, R.</dc:creator>
<dc:creator>Newcombe, J.</dc:creator>
<dc:creator>Greensmith, L.</dc:creator>
<dc:creator>Devine, H.</dc:creator>
<dc:creator>Taha, D. M.</dc:creator>
<dc:creator>Tyzack, G. E.</dc:creator>
<dc:creator>Hagemann, C.</dc:creator>
<dc:date>2020-08-17</dc:date>
<dc:identifier>doi:10.1101/2020.08.17.253773</dc:identifier>
<dc:title><![CDATA[Automated and unbiased classification of motor neuron phenotypes with single cell resolution in ALS tissue.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.08.18.256164v1?rss=1">
<title>
<![CDATA[
Amino acids whose intracellular levels change most during aging alter chronological lifespan of fission yeast 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.18.256164v1?rss=1"
</link>
<description><![CDATA[
Amino acid deprivation or supplementation can affect cellular and organismal lifespan, but we know little about the role of concentration changes in free, intracellular amino acids during aging. Here, we determine free amino-acid levels during chronological aging of non-dividing fission yeast cells. We compare wild-type with long-lived mutant cells that lack the Pka1 protein of the protein kinase A signalling pathway. In wild-type cells, total amino-acid levels decrease during aging, but much less so in pka1 mutants. Two amino acids strongly change as a function of age: glutamine decreases, especially in wild-type cells, while aspartate increases, especially in pka1 mutants. Supplementation of glutamine is sufficient to extend the chronological lifespan of wild-type but not of pka1{Delta} cells. Supplementation of aspartate, on the other hand, shortens the lifespan of pka1{Delta} but not of wild-type cells. Our results raise the possibility that certain amino acids are biomarkers of aging, and their concentrations during aging can promote or limit cellular lifespan.
]]></description>
<dc:creator>Rallis, C.</dc:creator>
<dc:creator>Mulleder, M.</dc:creator>
<dc:creator>Smith, G.</dc:creator>
<dc:creator>Au, Y. Z.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:creator>Bahler, J.</dc:creator>
<dc:date>2020-08-19</dc:date>
<dc:identifier>doi:10.1101/2020.08.18.256164</dc:identifier>
<dc:title><![CDATA[Amino acids whose intracellular levels change most during aging alter chronological lifespan of fission yeast]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.08.15.250589v1?rss=1">
<title>
<![CDATA[
Utilizing Computational Machine Learning Tools to Understand Immunogenic Breadth in the Context of a CD8 T-Cell Mediated HIV Response 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.15.250589v1?rss=1"
</link>
<description><![CDATA[
Predictive models are becoming more and more commonplace as tools for candidate antigen discovery to meet the challenges of enabling epitope mapping of cohorts with diverse HLA properties. Here we build on the concept of using two key parameters, diversity metric of the HLA profile of individuals within a population and consideration of sequence diversity in the context of an individuals CD8 T-cell immune repertoire to assess the HIV proteome for defined regions of immunogenicity. Using this approach, Analysis of HLA adaptation and functional immunogenicity data enabled the identification of regions within the proteome that offer significant conservation, HLA recognition within a population, low prevalence of HLA adaptation and demonstrated immunogenicity. We believe this unique and novel approach to vaccine design that, in combination with in vitro functional assays, offers a bespoke pipeline for expedited and rational CD8 T-cell vaccine design for HIV and potentially other pathogens with the potential for both global and local coverage.
]]></description>
<dc:creator>McGowan, E.</dc:creator>
<dc:creator>Rosenthal, R.</dc:creator>
<dc:creator>Fiore-Gartland, A.</dc:creator>
<dc:creator>Macharia, G.</dc:creator>
<dc:creator>Balinda, S.</dc:creator>
<dc:creator>Kapaata, A.</dc:creator>
<dc:creator>Muok, E.</dc:creator>
<dc:creator>Umviligihozo, G.</dc:creator>
<dc:creator>Dalel, J.</dc:creator>
<dc:creator>Streatfield, C.</dc:creator>
<dc:creator>Coutinho, H.</dc:creator>
<dc:creator>Monaco, D.</dc:creator>
<dc:creator>Morrison, D.</dc:creator>
<dc:creator>Yue, L.</dc:creator>
<dc:creator>Hunter, E.</dc:creator>
<dc:creator>Nielsen, M.</dc:creator>
<dc:creator>Gilmour, J.</dc:creator>
<dc:creator>Hare, J.</dc:creator>
<dc:date>2020-08-15</dc:date>
<dc:identifier>doi:10.1101/2020.08.15.250589</dc:identifier>
<dc:title><![CDATA[Utilizing Computational Machine Learning Tools to Understand Immunogenic Breadth in the Context of a CD8 T-Cell Mediated HIV Response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.08.22.261263v1?rss=1">
<title>
<![CDATA[
Active Perception during Angiogenesis: Filopodia speed up Notch selection of tip cells in silico and in vivo 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.22.261263v1?rss=1"
</link>
<description><![CDATA[
How do cells make efficient collective decisions during tissue morphogenesis? Humans and other organisms utilize feedback between movement and sensing known as  sensorimotor coordination or  active perception to inform behaviour, but active perception has not before been investigated at a cellular level within organs. Here we provide the first proof of concept in silico/in vivo study demonstrating that filopodia (actin-rich, dynamic, finger like cell-membrane protrusions) play an unexpected role in speeding up collective endothelial decisions during the time-constrained process of  tip cell selection during blood vessel formation (angiogenesis).

We first validate simulation predictions in vivo with live imaging of zebrafish intersegmental vessel growth. Further simulation studies then indicate the effect is due to the coupled positive feedback between movement and sensing on filopodia conferring a bistable switch-like property to Notch lateral inhibition, ensuring tip selection is a rapid and robust process. We then employ measures from computational neuroscience to assess whether filopodia function as a primitive ( basal) form of active perception and find evidence in support. By viewing cell behaviour in tissues through the  basal cognitive lens we acquire a fresh perspective on not only the well-studied tip cell selection process, revealing a hidden, yet vital, time-keeping role for filopodia, but on how to interpret and understand cell behaviour in general, opening up a myriad of new and exciting research directions.
]]></description>
<dc:creator>Zakirov, B.</dc:creator>
<dc:creator>Charalambous, G.</dc:creator>
<dc:creator>Aspalter, I. M.</dc:creator>
<dc:creator>Van-Vuuren, K.</dc:creator>
<dc:creator>Mead, T.</dc:creator>
<dc:creator>Harrington, K.</dc:creator>
<dc:creator>Thuret, R.</dc:creator>
<dc:creator>Ravasz Regan, E.</dc:creator>
<dc:creator>Herbert, S. P.</dc:creator>
<dc:creator>Bentley, K.</dc:creator>
<dc:date>2020-08-23</dc:date>
<dc:identifier>doi:10.1101/2020.08.22.261263</dc:identifier>
<dc:title><![CDATA[Active Perception during Angiogenesis: Filopodia speed up Notch selection of tip cells in silico and in vivo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.08.24.258657v1?rss=1">
<title>
<![CDATA[
MRE11 is crucial for malaria transmission and its absence affects expression of interconnected networks of key genes essential for life 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.24.258657v1?rss=1"
</link>
<description><![CDATA[
The Meiotic Recombination 11 protein (MRE11) plays a key role in DNA damage response and maintenance of genome stability. However, little is known about its function during development of the malaria parasite Plasmodium. Here, we present a functional, ultrastructural and transcriptomic analysis of Plasmodium MRE11 during its life-cycle in both mammalian and mosquito vector hosts. Genetic disruption of Plasmodium berghei mre11 (PbMRE11) results in significant retardation of oocyst development in the mosquito midgut associated with cytoplasmic and nuclear degeneration, along with concomitant ablation of sporogony and subsequent parasite transmission. Further, absence of PbMRE11 results in significant transcriptional downregulation of genes involved in key interconnected biological processes that are fundamental to all eukaryotic life including ribonucleoprotein biogenesis, spliceosome function and iron-sulphur cluster assembly. Overall, our study provides a comprehensive functional analysis of MRE11s role in Plasmodium development during the mosquito stages and offers a potential target for therapeutic intervention during malaria parasite transmission.
]]></description>
<dc:creator>Guttery, D.</dc:creator>
<dc:creator>Ramaprasad, A.</dc:creator>
<dc:creator>Ferguson, D.</dc:creator>
<dc:creator>Zeeshan, M.</dc:creator>
<dc:creator>Pandey, R.</dc:creator>
<dc:creator>Brady, D.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:creator>Pain, A.</dc:creator>
<dc:creator>Tewari, R.</dc:creator>
<dc:date>2020-08-24</dc:date>
<dc:identifier>doi:10.1101/2020.08.24.258657</dc:identifier>
<dc:title><![CDATA[MRE11 is crucial for malaria transmission and its absence affects expression of interconnected networks of key genes essential for life]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.08.24.264689v1?rss=1">
<title>
<![CDATA[
An essential amino acid synchronises malaria parasite development with daily host rhythms 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.24.264689v1?rss=1"
</link>
<description><![CDATA[
Rapid asexual replication of blood stage malaria parasites is responsible for the severity of disease symptoms and fuels the production of transmission forms. That malaria parasite species coordinate their cycles of asexual replication with daily rhythms of their host was discovered in the Hippocratic era, but how and why this occurs is enigmatic. Here, we demonstrate that the Plasmodium chabaudis schedule for asexual replication can be orchestrated by a isoleucine, metabolite provided to the parasite in periodic manner due to the hosts rhythmic intake of food. First, we identify nutrients with daily rhythms in the blood that match the timing of rhythms in both host feeding and the developmental schedule of asexually replicating parasites. We hypothesise that if parasites set their own developmental schedule, they should use a time-of-day cue that is a factor they cannot generate endogenously at any time-of-day, or scavenge in a round-the-clock manner. Our large-scale metabolomics experiment reveals that only one metabolite - the amino acid isoleucine - fits these criteria. Second, further experiments reveal that parasites alter the developmental schedule of asexual stages in response to isoleucine provision and withdrawal in the manner consistent with it acting as a time-cue. Specifically, parasites respond to isoleucine loss by slowing development. This is a parasite strategy rather than the consequences of an imposed constraint, because unlike when parasites are deprived of other essential nutrients, they suffer no apparent costs in the absence of isoleucine. Overall, our data suggest parasites can use the daily rhythmicity of blood-isoleucine concentration to synchronise asexual development with the availability of isoleucine, and potentially other resources, that arrive in the blood in a periodic manner due to the hosts daily feeding-fasting cycle. Identifying both how and why parasites keep time opens avenues for interventions; interfering with the parasites time-keeping mechanism may stall replication, increasing the efficacy of drugs and immune responses, and could also prevent parasites from entering dormancy to tolerate drugs.
]]></description>
<dc:creator>Prior, K. F.</dc:creator>
<dc:creator>Middleton, B.</dc:creator>
<dc:creator>Owolabi, A. T.</dc:creator>
<dc:creator>Westwood, M. L.</dc:creator>
<dc:creator>Holland, J.</dc:creator>
<dc:creator>O'Donnell, A. J.</dc:creator>
<dc:creator>Blackman, M. J.</dc:creator>
<dc:creator>Skene, D. J.</dc:creator>
<dc:creator>Reece, S. E.</dc:creator>
<dc:date>2020-08-24</dc:date>
<dc:identifier>doi:10.1101/2020.08.24.264689</dc:identifier>
<dc:title><![CDATA[An essential amino acid synchronises malaria parasite development with daily host rhythms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.08.25.266403v1?rss=1">
<title>
<![CDATA[
An Integrative Analysis of the Age-Associated Genomic, Transcriptomic and Epigenetic Landscape across Cancers 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.25.266403v1?rss=1"
</link>
<description><![CDATA[
Age is the most important risk factor for cancer, as cancer incidence and mortality increase with age. However, how molecular alterations in tumours differ among patients of different age remains largely unexplored. Here, using data from The Cancer Genome Atlas, we comprehensively characterised genomic, transcriptomic and epigenetic alterations in relation to patients age across cancer types. We showed that tumours from older patients present an overall increase in genomic instability, somatic copy-number alterations (SCNAs) and somatic mutations. Age-associated SCNAs and mutations were identified in several cancer-driver genes across different cancer types. The largest age-related genomic differences were found in gliomas and endometrial cancer. We identified age-related global transcriptomic changes and demonstrated that these genes are controlled by age-associated DNA methylation changes. This study provides a comprehensive view of age-associated alterations in cancer and underscores age as an important factor to consider in cancer research and clinical practice.
]]></description>
<dc:creator>Chatsirisupachai, K.</dc:creator>
<dc:creator>Lesluyes, T.</dc:creator>
<dc:creator>Paraoan, L.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>de Magalhaes, J. P.</dc:creator>
<dc:date>2020-08-25</dc:date>
<dc:identifier>doi:10.1101/2020.08.25.266403</dc:identifier>
<dc:title><![CDATA[An Integrative Analysis of the Age-Associated Genomic, Transcriptomic and Epigenetic Landscape across Cancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.08.21.261073v1?rss=1">
<title>
<![CDATA[
T-bet fate mapping identifies a novel ILC1-ILC2 subset in vivo 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.21.261073v1?rss=1"
</link>
<description><![CDATA[
Innate lymphoid cells (ILC) play a critical role in regulating immune responses at mucosal surfaces. Various subsets exist resembling T cell lineages defined by the expression of specific transcription factors. Thus, T-bet is expressed in ILC1 and Th1 cells. In order to further understand the functional roles of T-bet in ILC, we generated a fate-mapping mouse model that permanently marks cells and their progeny that are expressing, or have ever expressed T-bet. Here we have identified and characterised a novel ILC with characteristics of ILC1 and ILC2 that are "fate-mapped" for T-bet expression and arise early in neonatal life prior to establishment of a mature microbiome. These ILC1-ILC2 cells are critically dependent on T-bet and are able to express type 1 and type 2 cytokines at steady state, but not in the context of inflammation. These findings refine our understanding of ILC lineage regulation and stability and have important implications for the understanding of ILC biology at mucosal surfaces.

SUMMARYInnate lymphoid cells (ILC) play a critical role in regulating immune responses at mucosal surfaces. Three distinct ILC groups have been described according to expression of subset defining transcription factors and other markers. In this study we characterize a novel ILC subset with characteristics of group 1 and group 2 ILC in vivo.
]]></description>
<dc:creator>SCHROEDER, J.-H.</dc:creator>
<dc:creator>Garrido-Mesa, N.</dc:creator>
<dc:creator>Zabinski, T.</dc:creator>
<dc:creator>Gallagher, A.</dc:creator>
<dc:creator>Campbell, L.</dc:creator>
<dc:creator>Roberts, L.</dc:creator>
<dc:creator>Stolarczyk, E.</dc:creator>
<dc:creator>Beattie, G.</dc:creator>
<dc:creator>Lo, J.</dc:creator>
<dc:creator>Iseppon, A.</dc:creator>
<dc:creator>Moreira Heliodoro, C.</dc:creator>
<dc:creator>Reis, R.</dc:creator>
<dc:creator>Jenner, R.</dc:creator>
<dc:creator>Lavender, P.</dc:creator>
<dc:creator>Howard, J.</dc:creator>
<dc:creator>Grencis, R.</dc:creator>
<dc:creator>Helmby, H.</dc:creator>
<dc:creator>Neves, J.</dc:creator>
<dc:creator>Lord, G.</dc:creator>
<dc:date>2020-08-22</dc:date>
<dc:identifier>doi:10.1101/2020.08.21.261073</dc:identifier>
<dc:title><![CDATA[T-bet fate mapping identifies a novel ILC1-ILC2 subset in vivo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.08.28.267526v1?rss=1">
<title>
<![CDATA[
Plasmablast-derived antibody response to acute SARS-CoV-2 infection in humans 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.28.267526v1?rss=1"
</link>
<description><![CDATA[
Serological and plasmablast responses and plasmablast-derived IgG monoclonal antibodies (MAbs) have been analysed in three COVID-19 patients with different clinical severities. Potent humoral responses were detected within 3 weeks of onset of illness in all patients and the serological titre was elicited soon after or concomitantly with peripheral plasmablast response. An average of 13.7% and 13.0% of plasmablast-derived MAbs were reactive with virus spike glycoprotein or nucleocapsid, respectively. A subset of anti-spike (10 of 32) and over half of anti-nucleocapsid (19 of 35) antibodies cross-reacted with other betacoronaviruses tested and harboured extensive somatic mutations, indicative of an expansion of memory B cells upon SARS-CoV-2 infection. Fourteen of 32 anti-spike MAbs, including five anti-RBD, three anti-non-RBD S1 and six anti-S2, neutralised wild-type SARS-CoV-2 in independent assays. Anti-RBD MAbs were further grouped into four cross-inhibiting clusters, of which six antibodies from three separate clusters blocked the binding of RBD to ACE2 and five were neutralising. All ACE2-blocking anti-RBD antibodies were isolated from two patients with prolonged fever, which is compatible with substantial ACE2-blocking response in their sera. At last, the identification of non-competing pairs of neutralising antibodies would offer potential templates for the development of prophylactic and therapeutic agents against SARS-CoV-2.
]]></description>
<dc:creator>Huang, K.-Y. A.</dc:creator>
<dc:creator>Tan, T.</dc:creator>
<dc:creator>Chen, T.-H.</dc:creator>
<dc:creator>Huang, C.-G.</dc:creator>
<dc:creator>Harvey, R.</dc:creator>
<dc:creator>Hussain, S.</dc:creator>
<dc:creator>Chen, C.-P.</dc:creator>
<dc:creator>Harding, A.</dc:creator>
<dc:creator>Gilbert-Jaramillo, J.</dc:creator>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Knight, M.</dc:creator>
<dc:creator>Schimanski, L.</dc:creator>
<dc:creator>Shih, S.-R.</dc:creator>
<dc:creator>Lin, Y.-C.</dc:creator>
<dc:creator>Cheng, C.-Y.</dc:creator>
<dc:creator>Cheng, S.-H.</dc:creator>
<dc:creator>Huang, Y.-C.</dc:creator>
<dc:creator>Lin, T.-Y.</dc:creator>
<dc:creator>Rahikainen, R.</dc:creator>
<dc:creator>Howarth, M.</dc:creator>
<dc:creator>Jan, J.-T.</dc:creator>
<dc:creator>Ma, C.</dc:creator>
<dc:creator>James, W. S.</dc:creator>
<dc:creator>Daniels, R.</dc:creator>
<dc:creator>McCauley, J.</dc:creator>
<dc:creator>Rijal, P.</dc:creator>
<dc:creator>Townsend, A.</dc:creator>
<dc:date>2020-08-28</dc:date>
<dc:identifier>doi:10.1101/2020.08.28.267526</dc:identifier>
<dc:title><![CDATA[Plasmablast-derived antibody response to acute SARS-CoV-2 infection in humans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.08.31.275701v1?rss=1">
<title>
<![CDATA[
A COVID-19 vaccine candidate using SpyCatcher multimerization of the SARS-CoV-2 spike protein receptor-binding domain induces potent neutralising antibody responses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.31.275701v1?rss=1"
</link>
<description><![CDATA[
There is dire need for an effective and affordable vaccine against SARS-CoV-2 to tackle the ongoing pandemic. In this study, we describe a modular virus-like particle vaccine candidate displaying the SARS-CoV-2 spike glycoprotein receptor-binding domain (RBD) using SpyTag/SpyCatcher technology (RBD-SpyVLP). Low doses of RBD-SpyVLP in a prime-boost regimen induced a strong neutralising antibody response in mice and pigs that was superior to convalescent human sera. We evaluated antibody quality using ACE2 blocking and neutralisation of cell infection by pseudovirus or wild-type SARS-CoV-2. Using competition assays with a monoclonal antibody panel, we showed that RBD-SpyVLP induced a polyclonal antibody response that recognised all key epitopes on the RBD, reducing the likelihood of selecting neutralisation-escape mutants. The induction of potent and polyclonal antibody responses by RBD-SpyVLP provides strong potential to address clinical and logistic challenges of the COVID-19 pandemic. Moreover, RBD-SpyVLP is highly resilient, thermostable and can be lyophilised without losing immunogenicity, to facilitate global distribution and reduce cold-chain dependence.
]]></description>
<dc:creator>Tan, T. K.</dc:creator>
<dc:creator>Rijal, P.</dc:creator>
<dc:creator>Rahikainen, R.</dc:creator>
<dc:creator>Keeble, A.</dc:creator>
<dc:creator>Schimanski, L.</dc:creator>
<dc:creator>Hussain, S.</dc:creator>
<dc:creator>Harvey, R.</dc:creator>
<dc:creator>Hayes, J.</dc:creator>
<dc:creator>Edwards, J.</dc:creator>
<dc:creator>McLean, R.</dc:creator>
<dc:creator>Martini, V.</dc:creator>
<dc:creator>Pedrera, M.</dc:creator>
<dc:creator>Thakur, N.</dc:creator>
<dc:creator>Conceicao, C.</dc:creator>
<dc:creator>Dietrich, I.</dc:creator>
<dc:creator>Shelton, H.</dc:creator>
<dc:creator>Ludi, A. B.</dc:creator>
<dc:creator>Wilsden, G.</dc:creator>
<dc:creator>Browning, C.</dc:creator>
<dc:creator>Zagrajek, A.</dc:creator>
<dc:creator>Bialy, D.</dc:creator>
<dc:creator>Bhat, S.</dc:creator>
<dc:creator>Stevenson-Leggett, P.</dc:creator>
<dc:creator>Hollinghurst, P.</dc:creator>
<dc:creator>Tully, M.</dc:creator>
<dc:creator>Moffat, K.</dc:creator>
<dc:creator>Chiu, C.</dc:creator>
<dc:creator>Waters, R.</dc:creator>
<dc:creator>Gray, A.</dc:creator>
<dc:creator>Azhar, M.</dc:creator>
<dc:creator>Mioulet, V.</dc:creator>
<dc:creator>Newman, J.</dc:creator>
<dc:creator>Asfor, A. S.</dc:creator>
<dc:creator>Burman, A.</dc:creator>
<dc:creator>Crossley, S.</dc:creator>
<dc:creator>Hammond, J.</dc:creator>
<dc:creator>Tchilian, E.</dc:creator>
<dc:creator>Charleston, B.</dc:creator>
<dc:creator>Bailey, D.</dc:creator>
<dc:creator>Tuthill, T. J.</dc:creator>
<dc:creator>Graham, S.</dc:creator>
<dc:creator>Malinauskas, T.</dc:creator>
<dc:creator>Huo, J.</dc:creator>
<dc:creator>Tree, J.</dc:creator>
<dc:creator>Buttigieg, K.</dc:creator>
<dc:creator>Owens, R.</dc:creator>
<dc:creator></dc:creator>
<dc:date>2020-08-31</dc:date>
<dc:identifier>doi:10.1101/2020.08.31.275701</dc:identifier>
<dc:title><![CDATA[A COVID-19 vaccine candidate using SpyCatcher multimerization of the SARS-CoV-2 spike protein receptor-binding domain induces potent neutralising antibody responses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.08.26.267864v1?rss=1">
<title>
<![CDATA[
Modulation of the cAMP levels with a conserved actinobacteria phosphodiesterase enzyme reduces antimicrobial tolerance in mycobacteria 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.26.267864v1?rss=1"
</link>
<description><![CDATA[
Antimicrobial tolerance (AMT) is the gateway to the development of antimicrobial resistance (AMR) and is therefore a major issue that needs to be addressed.

The second messenger cyclic-AMP (cAMP), which is conserved across all taxa, is involved in propagating signals from environmental stimuli and converting these into a response. In bacteria, such as M. tuberculosis, P. aeruginosa, V. cholerae and B. pertussis, cAMP has been implicated in virulence, metabolic regulation and gene expression. However, cAMP signalling in mycobacteria is particularly complex due to the redundancy of adenylate cyclases, which are enzymes that catalyse the formation of cAMP from ATP, and the poor activity of the only known phosphodiesterase (PDE) enzyme, which degrades cAMP into 5- AMP.

Based on these two features, the modulation of this system with the aim of investigating cAMP signalling and its involvement in AMT in mycobacteria id difficult.

To address this pressing need, we identified a new cAMP-degrading phosphodiesterase enzyme (Rv1339) and used it to significantly decrease the intrabacterial levels of cAMP in mycobacteria. This analysis revealed that this enzyme increased the antimicrobial susceptibility of M. smegmatis mc2155. Using a combination of metabolomics, RNA-sequencing, antimicrobial susceptibility assays and bioenergetics analysis, we were able to characterize the molecular mechanism underlying this increased susceptibility.

This work represents an important milestone showing that the targeting of cAMP signalling is a promising new avenue for antimicrobial development and expands our understanding of cAMP signalling in mycobacteria.
]]></description>
<dc:creator>Thomson, M.</dc:creator>
<dc:creator>Nunta, K.</dc:creator>
<dc:creator>Cheyne, A.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Garza-Garcia, A.</dc:creator>
<dc:creator>Larrouy-Maumus, G.</dc:creator>
<dc:date>2020-08-26</dc:date>
<dc:identifier>doi:10.1101/2020.08.26.267864</dc:identifier>
<dc:title><![CDATA[Modulation of the cAMP levels with a conserved actinobacteria phosphodiesterase enzyme reduces antimicrobial tolerance in mycobacteria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.09.03.269555v1?rss=1">
<title>
<![CDATA[
Training a neural network to learn other dimensionality reduction removes data size restrictions in bioinformatics and provides a new route to exploring data representations 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.09.03.269555v1?rss=1"
</link>
<description><![CDATA[
High dimensionality omics and hyperspectral imaging datasets present difficult challenges for feature extraction and data mining due to huge numbers of features that cannot be simultaneously examined. The sample numbers and variables of these methods are constantly growing as new technologies are developed, and computational analysis needs to evolve to keep up with growing demand. Current state of the art algorithms can handle some routine datasets but struggle when datasets grow above a certain size. We present a training deep learning via neural networks on non-linear dimensionality reduction, in particular t-distributed stochastic neighbour embedding (t-SNE), to overcome prior limitations of these methods.

One Sentence SummaryAnalysis of prohibitively large datasets by combining deep learning via neural networks with non-linear dimensionality reduction.
]]></description>
<dc:creator>Dexter, A.</dc:creator>
<dc:creator>Thomas, S. A.</dc:creator>
<dc:creator>Steven, R. T.</dc:creator>
<dc:creator>Robinson, K. N.</dc:creator>
<dc:creator>Taylor, A. J.</dc:creator>
<dc:creator>Elia, E.</dc:creator>
<dc:creator>Nikula, C.</dc:creator>
<dc:creator>Campbell, A. D.</dc:creator>
<dc:creator>Panina, Y.</dc:creator>
<dc:creator>Najumudeen, A. K.</dc:creator>
<dc:creator>Murta, T.</dc:creator>
<dc:creator>Yan, B.</dc:creator>
<dc:creator>Grabowski, P.</dc:creator>
<dc:creator>Hamm, G.</dc:creator>
<dc:creator>Swales, J.</dc:creator>
<dc:creator>Gilmore, I.</dc:creator>
<dc:creator>Yuneva, M.</dc:creator>
<dc:creator>Goodwin, R. J. A.</dc:creator>
<dc:creator>Barry, S.</dc:creator>
<dc:creator>Sansom, O. J.</dc:creator>
<dc:creator>Takats, Z.</dc:creator>
<dc:creator>Bunch, J.</dc:creator>
<dc:date>2020-09-03</dc:date>
<dc:identifier>doi:10.1101/2020.09.03.269555</dc:identifier>
<dc:title><![CDATA[Training a neural network to learn other dimensionality reduction removes data size restrictions in bioinformatics and provides a new route to exploring data representations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.09.06.284679v1?rss=1">
<title>
<![CDATA[
Assessing the acoustic behaviour of Anopheles gambiae s.l. dsxF mutants: Implications for Vector Control 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.09.06.284679v1?rss=1"
</link>
<description><![CDATA[
BackgroundThe release of genetically modified mosquitoes which use gene-drive mechanisms to suppress reproduction in natural populations of Anopheles mosquitoes is one of the scientifically most promising methods for malaria transmission control. However, many scientific, regulatory and ethical questions remain before transgenic mosquitoes can be utilised in the field. Mutations which reduce an individuals reproductive success are likely to create strong selective pressures to evolve resistance. It is thus crucial that the targeted population collapses as rapidly and as completely as possible to reduce the available time for the emergence of drive-resistant mutations. At a behavioural level, this means that the gene-drive carrying mutants should be at least as (and ideally more) sexually attractive than the wildtype population they compete against. A key element in the copulatory negotiations of Anopheles mosquitoes is their acoustic courtship. We therefore analysed sound emissions and acoustic preference in a doublesex mutant previously used to successfully collapse caged colonies of Anopheles gambiae s.l..

MethodsThe flight tones produced by the beating of their wings form the signals for acoustic mating communication in Anopheles species. We assessed the acoustic impact of the disruption of a female-specific isoform of the doublesex gene (dsxF) on the wing beat frequency (WBF; measured as flight tone) of both males (XY) and females (XX) in homozygous dsxF- mutants (dsxF-/-), heterozygous dsxF- carriers (dsxF+/-) and G3  wildtype dsxF+ controls (dsxF+/+). To exclude non-genetic influences, we controlled for temperature and measured wing lengths for all experimental animals. We used a phonotaxis assay to test the acoustic preferences of mutant and control mosquitoes.

ResultsA previous study demonstrated an altered phenotype only for females homozygous for the disrupted dsx allele (dsxF-/-), who appear intersex. No phenotypic changes were observed for heterozygous carriers or males, suggesting that the female-specific dsxF allele is haplosufficient. We here identify significant, dose-dependent increases in the flight tones of both dsxF-/- and dsxF+/- females when compared to dsxF+/+ control females. Flight tone frequencies in all three female genotypes remained significantly lower than in males, however. When tested experimentally, males showed stronger phonotactic responses to the flight tones of control dsxF+/+ females. While flight tones from dsxF+/- and dsxF-/- females also elicited positive phonotactic behaviour in males, this was significantly reduced compared to responses to control tones. We found no evidence of phonotactic behaviour in any female genotype tested. None of the male genotypes displayed any deviations from the control condition.

ConclusionsA key prerequisite for copulation in anopheline mosquitoes is the phonotactic attraction of males towards female flight tones within large - spatially and acoustically crowded - mating swarms. Reductions in acoustic attractiveness of released mutant lines, as reported here for heterozygous dsxF+/- females, reduce the lines mating efficiency, and could consequently reduce the efficacy of the associated population control effort. Assessments of caged populations may not successfully reproduce the challenges posed by natural mating scenarios. We propose to amend existing testing protocols in order to more faithfully reflect the competitive conditions between a mutant line and the wildtype population it is meant to interact with. This should also include novel tests of  acoustic fitness. In line with previous studies, our findings confirm that disruption of the female-specific isoform dsxF has no effect on males; for some phenotypic traits, such as female flight tones, however, the effects of dsxF appear to be dose-dependent rather than haplosufficient.
]]></description>
<dc:creator>Su, M. P.</dc:creator>
<dc:creator>Georgiades, M.</dc:creator>
<dc:creator>Bagi, J.</dc:creator>
<dc:creator>Kyrou, K.</dc:creator>
<dc:creator>Crisanti, A.</dc:creator>
<dc:creator>Albert, J.</dc:creator>
<dc:date>2020-09-06</dc:date>
<dc:identifier>doi:10.1101/2020.09.06.284679</dc:identifier>
<dc:title><![CDATA[Assessing the acoustic behaviour of Anopheles gambiae s.l. dsxF mutants: Implications for Vector Control]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.09.07.285734v1?rss=1">
<title>
<![CDATA[
Plasmodium falciparum guanylyl cyclase-alpha and the activity of its appended P4-ATPase domain are essential for cGMP synthesis and blood stage egress 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.09.07.285734v1?rss=1"
</link>
<description><![CDATA[
In malaria parasites, guanylyl cyclases (GCs), which synthesise cyclic GMP (cGMP), are associated with a P4-ATPase-like domain in a unique bifunctional configuration. P4-ATPases generate membrane bilayer lipid asymmetry by translocating phospholipids from the outer to the inner leaflet. Here we investigate the role of Plasmodium falciparum guanylyl cyclase alpha (GC) and its associated P4-ATPase module, showing that asexual blood stage parasites lacking both the cyclase and P4-ATPase domains are unable to egress from host erythrocytes. GC-null parasites cannot synthesise cGMP, or mobilise calcium, a cGMP-dependent protein kinase (PKG)-driven requirement for egress. Using chemical complementation with a cGMP analogue and point mutagenesis of a crucial conserved residue within the P4-ATPase domain, we show that ATPase activity is up stream of and linked to cGMP synthesis. Collectively, our results demonstrate that GC is a critical regulator of PKG and that its associated P4-ATPase domain plays a primary role in generating cGMP for merozoite egress.
]]></description>
<dc:creator>Nofal, S. D.</dc:creator>
<dc:creator>Patel, A.</dc:creator>
<dc:creator>Blackman, M. J.</dc:creator>
<dc:creator>Flueck, C.</dc:creator>
<dc:creator>Baker, D. A.</dc:creator>
<dc:date>2020-09-07</dc:date>
<dc:identifier>doi:10.1101/2020.09.07.285734</dc:identifier>
<dc:title><![CDATA[Plasmodium falciparum guanylyl cyclase-alpha and the activity of its appended P4-ATPase domain are essential for cGMP synthesis and blood stage egress]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.09.07.285536v1?rss=1">
<title>
<![CDATA[
Generating single-sex litters: development of CRISPR-Cas9 genetic tools to produce all-male offspring 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.09.07.285536v1?rss=1"
</link>
<description><![CDATA[
Animals are extremely useful genetic tools in science and global resources in agriculture. However, a single sex is often required in surplus, and current genetic methods for producing all-female or all-male litters are inefficient. Using the mouse as a model, we developed a synthetic, two-part bicomponent strategy for generating all-male litters. We achieved this using CRISPR-Cas9 genome editing technology to generate large stable knock-ins on the autosomes and X chromosome. The bicomponent system functions via the sex-specific co-inheritance of a Cas9 transgene and an sgRNA transgene targeting the essential Topoisomerase 1 gene. This technology proved to be highly efficient in generating on-target mutations, resulting in embryonic lethality of the target sex. Our study is the first to successfully generate all-male mammalian litters using a CRISPR-Cas9 bicomponent system and provides great strides towards generating single-sex litters for laboratory or agricultural research.
]]></description>
<dc:creator>Douglas, C.</dc:creator>
<dc:creator>Maciulyte, V.</dc:creator>
<dc:creator>Zohren, J.</dc:creator>
<dc:creator>Snell, D. M.</dc:creator>
<dc:creator>Ojarikre, O. A.</dc:creator>
<dc:creator>Ellis, P. J.</dc:creator>
<dc:creator>Turner, J. M. A.</dc:creator>
<dc:date>2020-09-07</dc:date>
<dc:identifier>doi:10.1101/2020.09.07.285536</dc:identifier>
<dc:title><![CDATA[Generating single-sex litters: development of CRISPR-Cas9 genetic tools to produce all-male offspring]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.09.08.287797v1?rss=1">
<title>
<![CDATA[
Highly enriched hiPSC-derived midbrain dopaminergic neurons robustly models Parkinson's disease. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.09.08.287797v1?rss=1"
</link>
<description><![CDATA[
The development of human induced pluripotent stem cells (hiPSC) has greatly aided our ability to model neurodegenerative diseases. However, generation of midbrain dopaminergic (mDA) neurons is a major challenge and protocols are variable. Here, we developed a method to differentiate hiPSCs into enriched populations (>80%) of mDA neurons using only small molecules. We confirmed the identity of the mDA neurons using single-cell RNA-sequencing and detection of classical markers. Single-cell live imaging demonstrated neuronal calcium signalling and functional dopamine transport. Electrophysiology measures highlighted the ability to form synapses and networks in culture. Patient-specific hiPSC lines differentiated to produce functional mDA neurons that exhibit the hallmarks of synucleinopathy including: aggregate formation, oxidative stress as well as mitochondrial dysfunction and impaired lysosomal dynamics. In summary, we establish a robust differentiation paradigm to generate enriched mDA neurons from hiPSCs, which can be used to faithfully model key aspects of Parkinsons disease (PD), providing the potential to further elucidate molecular mechanisms contributing to disease development.
]]></description>
<dc:creator>Virdi, G. S.</dc:creator>
<dc:creator>Choi, M. L.</dc:creator>
<dc:creator>Yao, Z.</dc:creator>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>Athauda, D.</dc:creator>
<dc:creator>Melandri, D.</dc:creator>
<dc:creator>Sylantyev, S.</dc:creator>
<dc:creator>Abramov, A. Y.</dc:creator>
<dc:creator>Patani, R.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:date>2020-09-08</dc:date>
<dc:identifier>doi:10.1101/2020.09.08.287797</dc:identifier>
<dc:title><![CDATA[Highly enriched hiPSC-derived midbrain dopaminergic neurons robustly models Parkinson's disease.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.09.10.290866v1?rss=1">
<title>
<![CDATA[
The atypical Rho GTPase Rnd2 is critical for dentate granule neuron development and anxiety-like behavior during adult but not neonatal neurogenesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.09.10.290866v1?rss=1"
</link>
<description><![CDATA[
Despite the central role of Rho GTPases in neuronal development, their functions in adult hippocampal neurogenesis remain poorly explored. Here, by using a retrovirus-based loss-of-function approach in vivo, we show that the atypical Rho GTPase Rnd2 is crucial for the survival, positioning, somatodendritic morphogenesis and functional maturation of adult-born dentate granule neurons. Interestingly, most of these functions are specific to granule neurons generated during adulthood since the deletion of Rnd2 in neonatally-born granule neurons only affects dendritogenesis. In addition, suppression of Rnd2 in adult-born dentate granule neurons increases anxiety-like behaviour whereas its deletion in pups has no such effect, a finding supporting the adult neurogenesis hypothesis of anxiety disorders. Thus, our results provide mechanistic insight into the differential regulation of hippocampal neurogenesis during development and adulthood, and establishes a causal relationship between Rnd2 expression and anxiety.
]]></description>
<dc:creator>Kerloch, T.</dc:creator>
<dc:creator>Farrugia, F.</dc:creator>
<dc:creator>Maitre, M.</dc:creator>
<dc:creator>Terral, G.</dc:creator>
<dc:creator>Koehl, M.</dc:creator>
<dc:creator>Heng, J. I.-T.</dc:creator>
<dc:creator>Blanchard, M.</dc:creator>
<dc:creator>Doat, H.</dc:creator>
<dc:creator>Leste-Lasserre, T.</dc:creator>
<dc:creator>Goron, A.</dc:creator>
<dc:creator>Gonzales, D.</dc:creator>
<dc:creator>Guillemot, F.</dc:creator>
<dc:creator>Abrous, N.</dc:creator>
<dc:creator>Pacary, E.</dc:creator>
<dc:date>2020-09-10</dc:date>
<dc:identifier>doi:10.1101/2020.09.10.290866</dc:identifier>
<dc:title><![CDATA[The atypical Rho GTPase Rnd2 is critical for dentate granule neuron development and anxiety-like behavior during adult but not neonatal neurogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.09.09.289033v1?rss=1">
<title>
<![CDATA[
ImmunoCluster: A computational framework for the non-specialist to immune profile cellular heterogeneity using liquid and imaging mass, and flow cytometry datasets 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.09.09.289033v1?rss=1"
</link>
<description><![CDATA[
High dimensional cytometry is an innovative tool for immune monitoring in health and disease, it has provided novel insight into the underlying biology as well as biomarkers for a variety of diseases. However, the analysis of multiparametric "big data" usually requires specialist computational knowledge. Here we describe ImmunoCluster (https://github.com/kordastilab/ImmunoCluster) an R package for immune profiling cellular heterogeneity in high dimensional liquid and imaging mass cytometry, and flow cytometry data, designed to facilitate computational analysis by a non-specialist. The analysis framework implemented within ImmunoCluster is readily scalable to millions of cells and provides a variety of visualization and analytical approaches, as well as a rich array of plotting tools that can be tailored to users needs. The protocol consists of three core computational stages: 1, data import and quality control, 2, dimensionality reduction and unsupervised clustering; and 3, annotation and differential testing, all contained within an R-based open-source framework.
]]></description>
<dc:creator>Opzoomer, J. W.</dc:creator>
<dc:creator>Timms, J. A.</dc:creator>
<dc:creator>Blighe, K.</dc:creator>
<dc:creator>Mourikis, T. P.</dc:creator>
<dc:creator>Chapuis, N.</dc:creator>
<dc:creator>Bekoe, R.</dc:creator>
<dc:creator>Kareemaghay, S.</dc:creator>
<dc:creator>Nocerino, P.</dc:creator>
<dc:creator>Apollonio, B.</dc:creator>
<dc:creator>Ramsay, A. G.</dc:creator>
<dc:creator>Tavassoli, M.</dc:creator>
<dc:creator>Harrison, C.</dc:creator>
<dc:creator>Ciccarelli, F.</dc:creator>
<dc:creator>Parker, P.</dc:creator>
<dc:creator>Fontenay, M.</dc:creator>
<dc:creator>Barber, P.</dc:creator>
<dc:creator>Arnold, J. N.</dc:creator>
<dc:creator>Kordasti, S.</dc:creator>
<dc:date>2020-09-10</dc:date>
<dc:identifier>doi:10.1101/2020.09.09.289033</dc:identifier>
<dc:title><![CDATA[ImmunoCluster: A computational framework for the non-specialist to immune profile cellular heterogeneity using liquid and imaging mass, and flow cytometry datasets]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.09.10.291179v1?rss=1">
<title>
<![CDATA[
Acute depletion of METTL3 identifies a role for N6-methyladenosine in alternative intron/exon inclusion in the nascent transcriptome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.09.10.291179v1?rss=1"
</link>
<description><![CDATA[
RNA N6-methyladenosine (m6A) modification plays important roles in multiple aspects of RNA regulation. m6A is installed co-transcriptionally by the METTL3/14 complex, but its direct roles in RNA processing remain unclear. Here we investigate the presence of m6A in nascent RNA of mouse embryonic stem cells (mESCs). We find that around 10% m6A peaks are in introns, often close to 5-splice sites. RNA m6A peaks significantly overlap with RBM15 RNA binding sites and the histone modification H3K36me3. Interestingly, acute dTAG depletion of METTL3 reveals that inclusion of m6A-bearing alternative introns/exons in the nascent transcriptome is disrupted. For terminal or variable-length exons, m6A peaks are generally located upstream of a repressed 5-splice site, and downstream of an enhanced 5-splice site. Intriguingly, genes with the most immediate effects on splicing include several components of the m6A pathway, suggesting an autoregulatory function. Our findings demonstrate a direct crosstalk between m6A machinery and the regulation of RNA processing.
]]></description>
<dc:creator>Wei, G.</dc:creator>
<dc:creator>Almeida, M.</dc:creator>
<dc:creator>Pintacuda, G.</dc:creator>
<dc:creator>Coker, H.</dc:creator>
<dc:creator>Bowness, J. S.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Brockdorff, N.</dc:creator>
<dc:date>2020-09-11</dc:date>
<dc:identifier>doi:10.1101/2020.09.10.291179</dc:identifier>
<dc:title><![CDATA[Acute depletion of METTL3 identifies a role for N6-methyladenosine in alternative intron/exon inclusion in the nascent transcriptome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.09.11.292656v1?rss=1">
<title>
<![CDATA[
Telomere-to-telomere assembly of the genome of an individual Oikopleura dioica from Okinawa using Nanopore-based sequencing 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.09.11.292656v1?rss=1"
</link>
<description><![CDATA[
BackgroundThe larvacean Oikopleura dioica is an abundant tunicate plankton with the smallest (65-70 Mbp) non-parasitic, non-extremophile animal genome identified to date. Currently, there are two genomes available for the Bergen (OdB3) and Osaka (OSKA2016) O. dioica laboratory strains. Both assemblies have full genome coverage and high sequence accuracy. However, a chromosome-scale assembly has not yet been achieved.

ResultsHere, we present a chromosome-scale genome assembly (OKI2018_I69) of the Okinawan O. dioica produced using long-read Nanopore and short-read Illumina sequencing data from a single male, combined with Hi-C chromosomal conformation capture data for scaffolding. The OKI2018_I69 assembly has a total length of 64.3 Mbp distributed among 19 scaffolds. 99% of the assembly is in five megabase-scale scaffolds. We found telomeres on both ends of the two largest scaffolds, which represent assemblies of two fully contiguous autosomal chromosomes. Each of the other three large scaffolds have telomeres at one end only and we propose that they correspond to sex chromosomes split into a pseudo-autosomal region and X-specific or Y-specific regions. Indeed, these five scaffolds mostly correspond to equivalent linkage groups of OdB3, suggesting overall agreement in chromosomal organization between the two populations. At a more detailed level, the OKI2018_I69 assembly possesses similar genomic features in gene content and repetitive elements reported for OdB3. The Hi-C map suggests few reciprocal interactions between chromosome arms. At the sequence level, multiple genomic features such as GC content and repetitive elements are distributed differently along the short and long arms of the same chromosome.

ConclusionsWe show that a hybrid approach of integrating multiple sequencing technologies with chromosome conformation information results in an accurate de novo chromosome-scale assembly of O. dioicas highly polymorphic genome. This assembly will be a useful resource for genome-wide comparative studies between O. dioica and other species, as well as studies of chromosomal evolution in this lineage.
]]></description>
<dc:creator>Bliznina, A.</dc:creator>
<dc:creator>Masunaga, A.</dc:creator>
<dc:creator>Mansfield, M. J.</dc:creator>
<dc:creator>Tan, Y.</dc:creator>
<dc:creator>Liu, A. W.</dc:creator>
<dc:creator>West, C.</dc:creator>
<dc:creator>Rustagi, T.</dc:creator>
<dc:creator>Chien, H.-C.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Pichon, J.</dc:creator>
<dc:creator>Plessy, C.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:date>2020-09-12</dc:date>
<dc:identifier>doi:10.1101/2020.09.11.292656</dc:identifier>
<dc:title><![CDATA[Telomere-to-telomere assembly of the genome of an individual Oikopleura dioica from Okinawa using Nanopore-based sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.09.13.286047v1?rss=1">
<title>
<![CDATA[
A two-site flexible clamp mechanism for RET-GDNF-GFRα1 assembly reveals both conformational adaptation and strict geometric spacing 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.09.13.286047v1?rss=1"
</link>
<description><![CDATA[
RET receptor tyrosine kinase plays vital developmental and neuroprotective roles in metazoans. GDNF family ligands (GFLs) when bound to cognate GFR co-receptors recognise and activate RET stimulating its cytoplasmic kinase function. The principles for RET ligand-co-receptor recognition are incompletely understood. Here we report a crystal structure of the cadherin-like module (CLD1-4) from zebrafish RET revealing interdomain flexibility between CLD2-CLD3. Comparison with a cryo-EM structure of a ligand-engaged zebrafish RETECD-GDNF-GFR1 complex indicates conformational changes within a clade-specific CLD3 loop adjacent to co-receptor. Our observations indicate RET is a molecular clamp with a flexible calcium-dependent arm that adapts to different GFR co-receptors, while its rigid arm recognises a GFL dimer to align both membrane-proximal cysteine-rich domains. We also visualise linear arrays of RETECD-GDNF-GFR1 suggesting a conserved contact stabilises higher-order species. Our study reveals ligand-co-receptor recognition by RET involves both receptor plasticity and strict spacing of receptor dimers by GFL ligands.

HighlightsO_LICrystal structure of zebrafish RET cadherin-like module reveals conformational flexibility at the calcium-dependent CLD2-CLD3 interface
C_LIO_LIComparison of X-ray and cryo-EM structures indicate conformational differences between unliganded and liganded RET involving a clade-specific CLD3 loop
C_LIO_LIStrict spatial separation of RETECD C-termini is imposed by each cysteine-rich domain interaction with GFL dimer
C_LIO_LIDifferences in co-receptor engagement and higher-order ligand-bound RET complexes indicate potentially divergent signalling mechanisms
C_LI
]]></description>
<dc:creator>Adams, S.</dc:creator>
<dc:creator>Purkiss, A. G.</dc:creator>
<dc:creator>Knowles, P. P.</dc:creator>
<dc:creator>Nans, A.</dc:creator>
<dc:creator>Briggs, D. C.</dc:creator>
<dc:creator>Borg, A.</dc:creator>
<dc:creator>Earl, C. P.</dc:creator>
<dc:creator>Goodman, K. M.</dc:creator>
<dc:creator>Nawrotek, A.</dc:creator>
<dc:creator>Borg, A. J.</dc:creator>
<dc:creator>McIntosh, P. B.</dc:creator>
<dc:creator>Houghton, F. M.</dc:creator>
<dc:creator>Kjaer, S.</dc:creator>
<dc:creator>McDonald, N. Q.</dc:creator>
<dc:date>2020-09-13</dc:date>
<dc:identifier>doi:10.1101/2020.09.13.286047</dc:identifier>
<dc:title><![CDATA[A two-site flexible clamp mechanism for RET-GDNF-GFRα1 assembly reveals both conformational adaptation and strict geometric spacing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.09.15.297887v1?rss=1">
<title>
<![CDATA[
Single-molecule measurements reveal that PARP1 condenses DNA by loop formation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.09.15.297887v1?rss=1"
</link>
<description><![CDATA[
Poly(ADP-ribose) polymerase 1 (PARP1) is an abundant nuclear enzyme that plays important roles in DNA repair, chromatin organization and transcription regulation. Although binding and activation of PARP1 by DNA damage sites has been extensively studied, little is known about how PARP1 binds to long stretches of undamaged DNA and how it could shape chromatin architecture. Here, using a combination of single-molecule techniques including magnetic tweezers and atomic force microscopy, we show that PARP1 binds and condenses undamaged, kilobase-length DNA subject to sub-picoNewton mechanical forces. Decondensation by high force proceeds through a series of discrete increases in extension, indicating that PARP1 stabilizes loops of DNA. This model is supported by DNA braiding experiments which show that PARP1 can bind at the intersection of two separate DNA molecules. PARP inhibitors do not affect the level of condensation of undamaged DNA, but act to block condensation reversal for damaged DNA in the presence of NAD+. Our findings establish a mechanism for PARP1 in the organization of chromatin structure.
]]></description>
<dc:creator>Bell, N. A. W.</dc:creator>
<dc:creator>Haynes, P. J.</dc:creator>
<dc:creator>Brunner, K.</dc:creator>
<dc:creator>Maia de Oliveira, T.</dc:creator>
<dc:creator>Flocco, M.</dc:creator>
<dc:creator>Hoogenboom, B. W.</dc:creator>
<dc:creator>Molloy, J. E.</dc:creator>
<dc:date>2020-09-15</dc:date>
<dc:identifier>doi:10.1101/2020.09.15.297887</dc:identifier>
<dc:title><![CDATA[Single-molecule measurements reveal that PARP1 condenses DNA by loop formation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.09.17.301366v1?rss=1">
<title>
<![CDATA[
The eukaryotic replisome requires an additional helicase to disarm dormant replication origins 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.09.17.301366v1?rss=1"
</link>
<description><![CDATA[
Origins of eukaryotic DNA replication are  licensed during G1 phase of the cell cycle by loading the six related minichromosome maintenance (MCM) proteins into a double hexameric ring around double-stranded DNA. In S phase, some double hexamers (MCM DHs) are converted into active CMG (Cdc45-MCM-GINS) helicases which nucleate assembly of bidirectional replication forks. The remaining unfired MCM DHs act as  dormant origins to provide backup replisomes in the event of replication fork stalling. The fate of unfired MCM DHs during replication is unknown. Here we show that active replisomes cannot remove unfired MCM DHs. Instead, they are pushed ahead of the replisome where they prevent fork convergence during replication termination and replisome progression through nucleosomes. Pif1 helicase, together with the replisome, can remove unfired MCM DHs specifically from replicating DNA, allowing efficient replication and termination. Our results provide an explanation for how excess replication license is removed during S phase.
]]></description>
<dc:creator>Hill, J.</dc:creator>
<dc:creator>Eickhoff, P.</dc:creator>
<dc:creator>Drury, L.</dc:creator>
<dc:creator>Costa, A.</dc:creator>
<dc:creator>Diffley, J.</dc:creator>
<dc:date>2020-09-17</dc:date>
<dc:identifier>doi:10.1101/2020.09.17.301366</dc:identifier>
<dc:title><![CDATA[The eukaryotic replisome requires an additional helicase to disarm dormant replication origins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.09.18.303040v1?rss=1">
<title>
<![CDATA[
Primary sex determination in chickens depends on DMRT1 dosage, but gonadal sex does not determine secondary sexual characteristics in adult birds 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.09.18.303040v1?rss=1"
</link>
<description><![CDATA[
In birds, males are the homogametic sex (ZZ) and females the heterogametic sex (ZW), and primary sex determination is thought to depend on a sex chromosome gene dosage mechanism. Previous studies have suggested that the most likely sex-determinant is the Z chromosome gene DMRT1 (Doublesex and Mab-3 Related Transcription factor 1). To clarify this issue, we used a CRISPR-Cas9 based mono-allelic targeting approach and sterile surrogate hosts to generate birds with targeted mutations in the DMRT1 gene. The resulting chromosomally male (ZZ) chicken with a single functional copy of DMRT1 developed ovaries in place of testes, demonstrating the avian sex determining mechanism is based on DMRT1 dosage. These ZZ ovaries expressed typical female markers and showed clear evidence of follicular development. However, these ZZ adult birds with an ovary in place of testes were indistinguishable in appearance to wild type adult males, supporting the concept of cell-autonomous sex identity (CASI) in birds. In experiments where oestrogen synthesis was blocked in control ZW embryos, the resulting gonads developed as testes. In contrast, if oestrogen synthesis was blocked in ZW embryos that lacked DMRT1, the gonads invariably adopted an ovarian fate. Our analysis shows that DMRT1 is the key sex determination switch in birds and that it is essential for testis development, but that production of oestrogen is also a key factor in primary sex determination in chickens, and that this production is linked to DMRT1 expression.
]]></description>
<dc:creator>Ioannidis, J.</dc:creator>
<dc:creator>Taylor, G.</dc:creator>
<dc:creator>Zhao, D.</dc:creator>
<dc:creator>Liu, L.</dc:creator>
<dc:creator>Idoko-Akoh, A.</dc:creator>
<dc:creator>Gong, D.</dc:creator>
<dc:creator>Lovell-Badge, R.</dc:creator>
<dc:creator>Guioli, S.</dc:creator>
<dc:creator>McGrew, M.</dc:creator>
<dc:creator>Clinton, M.</dc:creator>
<dc:date>2020-09-19</dc:date>
<dc:identifier>doi:10.1101/2020.09.18.303040</dc:identifier>
<dc:title><![CDATA[Primary sex determination in chickens depends on DMRT1 dosage, but gonadal sex does not determine secondary sexual characteristics in adult birds]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.09.21.306522v1?rss=1">
<title>
<![CDATA[
MICAL2 acts through Arp3B isoform-specific Arp2/3 complexes to destabilize branched actin networks 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.09.21.306522v1?rss=1"
</link>
<description><![CDATA[
The Arp2/3 complex (Arp2, Arp3 and ARPC1-5) is essential to generate branched actin filament networks for many cellular processes. Human Arp3, ARPC1 and ARPC5 exist as two isoforms but the functional properties of Arp2/3 iso-complexes is largely unexplored. Here we show that Arp3B, but not Arp3 is subject to regulation by the methionine monooxygenase MICAL2, which is recruited to branched actin networks by coronin-1C. Although Arp3 and Arp3B iso-complexes promote actin assembly equally efficiently in vitro, they have different cellular properties. Arp3B turns over significantly faster than Arp3 within the network and upon its depletion actin turnover decreases. Substitution of Arp3B Met293 by Thr, the corresponding residue in Arp3 increases actin network stability, and conversely, replacing Arp3 Thr293 with Gln to mimic Met oxidation promotes network disassembly. Thus, MICAL2 regulates a subset of Arp2/3 complexes to control branched actin network disassembly.
]]></description>
<dc:creator>Galloni, C.</dc:creator>
<dc:creator>Carra, D.</dc:creator>
<dc:creator>Abella, J. V. G.</dc:creator>
<dc:creator>Kjaer, S.</dc:creator>
<dc:creator>Singaravelu, P.</dc:creator>
<dc:creator>Barry, D. J.</dc:creator>
<dc:creator>Kogata, N.</dc:creator>
<dc:creator>Guerin, C.</dc:creator>
<dc:creator>Blanchoin, L.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:date>2020-09-21</dc:date>
<dc:identifier>doi:10.1101/2020.09.21.306522</dc:identifier>
<dc:title><![CDATA[MICAL2 acts through Arp3B isoform-specific Arp2/3 complexes to destabilize branched actin networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.09.24.311613v1?rss=1">
<title>
<![CDATA[
Dual-view oblique plane microscopy (dOPM) 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.09.24.311613v1?rss=1"
</link>
<description><![CDATA[
We present a new folded dual-view oblique plane microscopy (OPM) technique termed dOPM that enables two orthogonal views of the sample to be obtained by translating a pair of tilted mirrors in refocussing space. Using a water immersion 40x 1.15 NA primary objective, deconvolved image volumes of 200 nm beads were measured to have full width at half maxima (FWHM) of 0.35{+/-}0.04 m and 0.39{+/-}0.02 m laterally and 0.81{+/-}0.07 m axially. The laterally integrated z-sectioning value was 1.33{+/-}0.45 m using light-sheet FWHM in the frames of the two views of 4.99{+/-}0.58 m and 4.89{+/-}0.63 m. To qualitatively demonstrate that the system can reduce shadow artefacts while providing a more isotropic resolution, a multi-cellular spheroid approximately 100 m in diameter was imaged.
]]></description>
<dc:creator>Sparks, H.</dc:creator>
<dc:creator>Dent, L.</dc:creator>
<dc:creator>Bakal, C.</dc:creator>
<dc:creator>Behrens, A.</dc:creator>
<dc:creator>Salbreux, G.</dc:creator>
<dc:creator>Dunsby, C.</dc:creator>
<dc:date>2020-09-25</dc:date>
<dc:identifier>doi:10.1101/2020.09.24.311613</dc:identifier>
<dc:title><![CDATA[Dual-view oblique plane microscopy (dOPM)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.09.28.316620v1?rss=1">
<title>
<![CDATA[
Evaluating the fitness of PA/I38T-substituted influenza A viruses with reduced baloxavir susceptibility in a competitive mixtures ferret model 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.09.28.316620v1?rss=1"
</link>
<description><![CDATA[
Baloxavir is approved in several countries for the treatment of uncomplicated influenza in otherwise-healthy and high-risk patients. Treatment-emergent viruses with reduced susceptibility to baloxavir have been detected in clinical trials, but the likelihood of widespread occurrence depends on replication capacity and onward transmission. We evaluated the fitness of A/H3N2 and A/H1N1pdm09 viruses with the polymerase acidic I38T-variant conferring reduced susceptibility to baloxavir relative to wild-type (WT) viruses, using a competitive mixture ferret model, recombinant viruses and patient-derived virus isolates. The A/H3N2 I38T virus showed a reduction in within-host fitness but comparable between-host fitness to the WT virus, while the A/H1N1pdm09 I38T virus had broadly similar within-host fitness but substantially lower between-host fitness. Although I38T viruses replicate and transmit between ferrets, our data suggest that viruses with this amino acid substitution have lower fitness relative to WT and this relative fitness cost was greater in A/H1N1pdm09 viruses than in A/H3N2 viruses.
]]></description>
<dc:creator>Lee, L. Y.</dc:creator>
<dc:creator>Zhou, J.</dc:creator>
<dc:creator>Koszalka, P.</dc:creator>
<dc:creator>Frise, R.</dc:creator>
<dc:creator>Farrukee, R.</dc:creator>
<dc:creator>Baba, K.</dc:creator>
<dc:creator>Miah, S.</dc:creator>
<dc:creator>Shishido, T.</dc:creator>
<dc:creator>Galiano, M.</dc:creator>
<dc:creator>Hashimoto, T.</dc:creator>
<dc:creator>Omoto, S.</dc:creator>
<dc:creator>Uehara, T.</dc:creator>
<dc:creator>Mifsud, E.</dc:creator>
<dc:creator>Collinson, N.</dc:creator>
<dc:creator>Kuhlbusch, K.</dc:creator>
<dc:creator>Clinch, B.</dc:creator>
<dc:creator>Wildum, S.</dc:creator>
<dc:creator>Barclay, W. S.</dc:creator>
<dc:creator>Hurt, A. C.</dc:creator>
<dc:date>2020-09-28</dc:date>
<dc:identifier>doi:10.1101/2020.09.28.316620</dc:identifier>
<dc:title><![CDATA[Evaluating the fitness of PA/I38T-substituted influenza A viruses with reduced baloxavir susceptibility in a competitive mixtures ferret model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.10.02.323873v1?rss=1">
<title>
<![CDATA[
LRRK2-dependent Rab GTPase phosphorylation in response to endolysosomal damage depends on macrophage differentiation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.10.02.323873v1?rss=1"
</link>
<description><![CDATA[
The Parkinsons Disease (PD) kinase LRRK2 is highly expressed in immune cells such as macrophages. In these cells, LRRK2 regulates innate immune pathways and it is activated after membrane damage leading to the phosphorylation of the Rab GTPases Rab8A and Rab10. Due to their wide-range of functions in immunity and tissue remodelling, macrophages in vivo are phenotypically heterogeneous. In vitro systems are used to differentiate these cells into diverse macrophage subsets to mimic the populations observed in vivo. M-CSF and GM-CSF differentiated human blood monocytes are often used to generate monocyte-derived macrophages as a model for tissue macrophages. However, how LRRK2 is activated in different macrophage subsets after membrane damage is unknown.

Here, we report that bone marrow derived macrophages and human monocyte-derived macrophages differentiated with either M-CSF or GM-CSF show different levels of LRRK2 activation after membrane damage. Notably, the membrane damaging agent LLOMe triggered LRRK2-dependent Rab8A and Rab10 phosphorylation primarily in GM-CSF differentiated macrophages. Moreover, LRRK2 and Rab8A were recruited to damaged endolysosomes in GM-CSF differentiated macrophages. Strikingly, GM-CSF differentiated macrophages recruited significantly more CHMP4B and Galectin-3 into damaged endolysosomes. These results suggest that LRRK2-regulated pathways of endolysosomal membrane damage and repair differ between macrophage subsets.
]]></description>
<dc:creator>Herbst, S.</dc:creator>
<dc:creator>Gutierrez, M. G.</dc:creator>
<dc:date>2020-10-02</dc:date>
<dc:identifier>doi:10.1101/2020.10.02.323873</dc:identifier>
<dc:title><![CDATA[LRRK2-dependent Rab GTPase phosphorylation in response to endolysosomal damage depends on macrophage differentiation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.09.30.306795v1?rss=1">
<title>
<![CDATA[
Differing total mRNA expression shapes the molecular and clinical phenotype of cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.09.30.306795v1?rss=1"
</link>
<description><![CDATA[
Cancers can vary greatly in their transcriptomes. In contrast to alterations in specific genes or pathways, differences in tumor cell total mRNA content have not been comprehensively assessed. Technical and analytical challenges have impeded examination of total mRNA expression at scale across cancers. To address this, we developed a model for quantifying tumor-specific total mRNA expression (TmS) from bulk sequencing data, which performs transcriptomic deconvolution while adjusting for mixed genomes. We used single-cell RNA sequencing data to demonstrate total mRNA expression as a feature of tumor phenotype. We estimated and validated TmS in 5,015 patients across 15 cancer types identifying significant inter-individual variability. At a pan-cancer level, high TmS is associated with increased risk of disease progression and death. Cancer type-specific patterns of genetic alterations, intra-tumor genetic heterogeneity, as well as pan-cancer trends in metabolic dysregulation and hypoxia contribute to TmS. Taken together, our results suggest that measuring cell-type specific total mRNA expression offers a broader perspective of tracking cancer transcriptomes, which has important biological and clinical implications.
]]></description>
<dc:creator>Cao, S.</dc:creator>
<dc:creator>Wang, J. R.</dc:creator>
<dc:creator>Ji, S.</dc:creator>
<dc:creator>Yang, P.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Montierth, M. D.</dc:creator>
<dc:creator>Shen, J. P.</dc:creator>
<dc:creator>Lee, J. J.</dc:creator>
<dc:creator>Guerrero, P. A.</dc:creator>
<dc:creator>Yu, K.</dc:creator>
<dc:creator>Livingstone, J.</dc:creator>
<dc:creator>Bhandari, V.</dc:creator>
<dc:creator>Hubert, S. M.</dc:creator>
<dc:creator>Daw, N. C.</dc:creator>
<dc:creator>Futreal, A.</dc:creator>
<dc:creator>Efstathiou, E.</dc:creator>
<dc:creator>Lim, B.</dc:creator>
<dc:creator>Viale, A.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Maitra, A.</dc:creator>
<dc:creator>Kopetz, S.</dc:creator>
<dc:creator>Campbell, P.</dc:creator>
<dc:creator>Speed, T. P.</dc:creator>
<dc:creator>Boutros, P. C.</dc:creator>
<dc:creator>Urbanucci, A.</dc:creator>
<dc:creator>Zhu, H.</dc:creator>
<dc:creator>Demeulemeester, J.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:date>2020-10-02</dc:date>
<dc:identifier>doi:10.1101/2020.09.30.306795</dc:identifier>
<dc:title><![CDATA[Differing total mRNA expression shapes the molecular and clinical phenotype of cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.10.04.325365v1?rss=1">
<title>
<![CDATA[
Somatic chromosomal number alterations affecting driver genes inform in-vitro and clinical drug response in high-grade serous ovarian cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.10.04.325365v1?rss=1"
</link>
<description><![CDATA[
The genomic complexity and heterogeneity of high-grade serous ovarian cancer (HGSOC) has hampered the realisation of successful therapies and effective personalised treatment is an unmet clinical need. Here we show that primary HGSOC spheroid models can be used to predict drug response and use them to demonstrate that somatic copy number alterations (SCNAs) in frequently amplified HGSOC cancer genes significantly correlate with gene expression and drug response. These genes are often located in areas of the genome with frequent clonal SCNAs. MYC chromosomal copy number is associated with ex-vivo and clinical response to paclitaxel and ex-vivo response to mTORC1/2 inhibition. Activation of the mTOR survival pathway in the context to MYC-amplified HGSOC is mostly due to increased prevalence of SCNAs in genes from the PI3K pathway. These results suggest that SCNAs encompassing driver genes could be used to inform therapeutic response in the context of clinical trials testing personalised medicines.
]]></description>
<dc:creator>Correia Martins, F.</dc:creator>
<dc:creator>Couturier, D.-L.</dc:creator>
<dc:creator>Santiago, I.</dc:creator>
<dc:creator>Sauer, C.</dc:creator>
<dc:creator>Vias, M.</dc:creator>
<dc:creator>Angelova, M.</dc:creator>
<dc:creator>Sanders, D.</dc:creator>
<dc:creator>Piskorz, A.</dc:creator>
<dc:creator>Hall, J.</dc:creator>
<dc:creator>Hosking, K.</dc:creator>
<dc:creator>Amirthanayagam, A.</dc:creator>
<dc:creator>Cosulich, S.</dc:creator>
<dc:creator>Carnevalli, L.</dc:creator>
<dc:creator>Davies, B.</dc:creator>
<dc:creator>Watkins, T. B. K.</dc:creator>
<dc:creator>Funingana, G.</dc:creator>
<dc:creator>Bolton, H.</dc:creator>
<dc:creator>Haldar, K.</dc:creator>
<dc:creator>Latimer, J.</dc:creator>
<dc:creator>Baldwin, P.</dc:creator>
<dc:creator>Crawford, R.</dc:creator>
<dc:creator>Eldridge, M.</dc:creator>
<dc:creator>Basu, B.</dc:creator>
<dc:creator>Jimenez-Linan, M.</dc:creator>
<dc:creator>McGranahan, N.</dc:creator>
<dc:creator>Litchfield, K.</dc:creator>
<dc:creator>Shah, S.</dc:creator>
<dc:creator>McNeish, I.</dc:creator>
<dc:creator>Caldas, C.</dc:creator>
<dc:creator>Evan, G.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Brenton, J. D.</dc:creator>
<dc:date>2020-10-04</dc:date>
<dc:identifier>doi:10.1101/2020.10.04.325365</dc:identifier>
<dc:title><![CDATA[Somatic chromosomal number alterations affecting driver genes inform in-vitro and clinical drug response in high-grade serous ovarian cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.10.07.330407v1?rss=1">
<title>
<![CDATA[
Mutations in SKI in Shprintzen-Goldberg syndrome lead to attenuated TGF-β responses through SKI stabilization 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.10.07.330407v1?rss=1"
</link>
<description><![CDATA[
Shprintzen-Goldberg syndrome (SGS) is a multisystemic connective tissue disorder, with considerable clinical overlap with Marfan and Loeys-Dietz syndromes. These syndromes have commonly been associated with enhanced TGF-{beta} signaling. In SGS patients, heterozygous point mutations have been mapped to the transcriptional corepressor SKI, which is a negative regulator of TGF-{beta} signaling that is rapidly degraded upon ligand stimulation. The molecular consequences of these mutations, however, are not understood. Here we use a combination of structural biology, genome editing and biochemistry to show that SGS mutations in SKI abolish its binding to phosphorylated SMAD2 and SMAD3. This results in stabilization of SKI and consequently attenuation of TGF-{beta} responses, in both knockin cells expressing an SGS mutation, and in fibroblasts from SGS patients. Thus, we reveal that SGS is associated with an attenuation of TGF-{beta}-induced transcriptional responses, and not enhancement, which has important implications for other Marfan-related syndromes.
]]></description>
<dc:creator>Gori, I.</dc:creator>
<dc:creator>George, R.</dc:creator>
<dc:creator>Purkiss, A.</dc:creator>
<dc:creator>Strohbuecker, S.</dc:creator>
<dc:creator>Randall, R. A.</dc:creator>
<dc:creator>Ogrodowicz, R.</dc:creator>
<dc:creator>Carmignac, V.</dc:creator>
<dc:creator>Faivre, L.</dc:creator>
<dc:creator>Joshi, D.</dc:creator>
<dc:creator>Kjaer, S.</dc:creator>
<dc:creator>Hill, C. S.</dc:creator>
<dc:date>2020-10-09</dc:date>
<dc:identifier>doi:10.1101/2020.10.07.330407</dc:identifier>
<dc:title><![CDATA[Mutations in SKI in Shprintzen-Goldberg syndrome lead to attenuated TGF-β responses through SKI stabilization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.10.08.331942v1?rss=1">
<title>
<![CDATA[
ES-62 suppression of arthritis reflects epigenetic rewiring of synovial fibroblasts to a joint-protective phenotype 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.10.08.331942v1?rss=1"
</link>
<description><![CDATA[
The parasitic worm product, ES-62 protects against collagen-induced arthritis, a mouse model of rheumatoid arthritis (RA) by suppressing the synovial fibroblast (SF) responses perpetuating inflammation and driving joint destruction. Such SF responses are shaped during disease progression by the inflammatory microenvironment of the joint that promotes remodelling of their epigenetic landscape, inducing an "aggressive" pathogenic SF phenotype. Critically, exposure to ES-62 in vivo induces a stably imprinted "safe" phenotype that exhibits responses more typical of healthy SFs. Surprisingly however, DNA methylome analysis reveals that rather than simply preventing the pathogenic rewiring of SFs, ES-62 induces further epigenetic remodelling, including targeting genes associated with ciliogenesis and differentiation, to program a distinct "protective" phenotype. Such unique behaviour signposts potential DNA methylation signatures predictive of pathogenesis and its resolution and hence, candidate mechanisms by which novel therapeutic interventions could prevent SFs from perpetuating joint inflammation and destruction in RA.
]]></description>
<dc:creator>Corbet, M.</dc:creator>
<dc:creator>Pineda, M. A.</dc:creator>
<dc:creator>Yang, K.</dc:creator>
<dc:creator>Tarafdar, A.</dc:creator>
<dc:creator>McGrath, S.</dc:creator>
<dc:creator>Nakagawa, R.</dc:creator>
<dc:creator>Lumb, F. E.</dc:creator>
<dc:creator>Harnett, W.</dc:creator>
<dc:creator>Harnett, M. M.</dc:creator>
<dc:date>2020-10-08</dc:date>
<dc:identifier>doi:10.1101/2020.10.08.331942</dc:identifier>
<dc:title><![CDATA[ES-62 suppression of arthritis reflects epigenetic rewiring of synovial fibroblasts to a joint-protective phenotype]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.10.15.340810v1?rss=1">
<title>
<![CDATA[
GIANI: open-source software for automated analysis of 3D microscopy images 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.10.15.340810v1?rss=1"
</link>
<description><![CDATA[
The study of cellular and developmental processes in physiologically relevant three-dimensional (3D) systems facilitates an understanding of mechanisms underlying cell fate, disease and injury. While cutting-edge microscopy technologies permit the routine acquisition of 3D datasets, there is currently a limited number of open-source software packages to analyse such images. Here we describe GIANI (djpbarry.github.io/Giani), new software for the analysis of 3D images, implemented as a plugin for the popular FIJI platform. The design primarily facilitates segmentation of nuclei and cells, followed by quantification of morphology and protein expression. GIANI enables routine and reproducible batch-processing of large numbers of images and also comes with scripting and command line tools, allowing users to incorporate its functionality into their own scripts and also run GIANI on a high-performance computing cluster. We demonstrate the utility of GIANI by quantifying cell morphology and protein expression in confocal images of mouse early embryos and by segmenting nuclei from light sheet microscopy images of the flour beetle embryo. We also validate the performance of the software using simulated data. More generally, we anticipate that GIANI will be a useful tool for researchers in a variety of biomedical fields.
]]></description>
<dc:creator>Barry, D. J.</dc:creator>
<dc:creator>Gerri, C.</dc:creator>
<dc:creator>Bell, D. M.</dc:creator>
<dc:creator>D'Antuono, R.</dc:creator>
<dc:creator>Niakan, K. K.</dc:creator>
<dc:date>2020-10-16</dc:date>
<dc:identifier>doi:10.1101/2020.10.15.340810</dc:identifier>
<dc:title><![CDATA[GIANI: open-source software for automated analysis of 3D microscopy images]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.10.15.341305v1?rss=1">
<title>
<![CDATA[
Computational and experimental analyses of mitotic chromosome formation pathways in fission yeast 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.10.15.341305v1?rss=1"
</link>
<description><![CDATA[
Underlying higher order chromatin organization are Structural Maintenance of Chromosomes (SMC) complexes, large protein rings that entrap DNA. The molecular mechanism by which SMC complexes organize chromatin is as yet incompletely understood. Two prominent models posit that SMC complexes actively extrude DNA loops (loop extrusion), or that they sequentially entrap two DNAs that come into proximity by Brownian motion (diffusion capture). To explore the implications of these two mechanisms, we perform biophysical simulations of a 3.76 Mb-long chromatin chain, the size of the long S. pombe chromosome I left arm. On it, the SMC complex condensin is modeled to perform loop extrusion or diffusion capture. We then compare computational to experimental observations of mitotic chromosome formation. Both loop extrusion and diffusion capture can result in native-like contact probability distributions. In addition, the diffusion capture model more readily recapitulates mitotic chromosome axis shortening and chromatin density enrichment. Diffusion capture can also explain why mitotic chromatin shows reduced, as well as more anisotropic, movements, features that lack support from loop extrusion. The condensin distribution within mitotic chromosomes, visualized by stochastic optical reconstruction microscopy (STORM), shows clustering predicted from diffusion capture. Our results inform the evaluation of current models of mitotic chromosome formation.
]]></description>
<dc:creator>Gerguri, T.</dc:creator>
<dc:creator>Fu, X.</dc:creator>
<dc:creator>Kakui, Y.</dc:creator>
<dc:creator>Khatri, B. S.</dc:creator>
<dc:creator>Barrington, C.</dc:creator>
<dc:creator>Bates, P. A.</dc:creator>
<dc:creator>Uhlmann, F.</dc:creator>
<dc:date>2020-10-16</dc:date>
<dc:identifier>doi:10.1101/2020.10.15.341305</dc:identifier>
<dc:title><![CDATA[Computational and experimental analyses of mitotic chromosome formation pathways in fission yeast]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.10.20.345306v1?rss=1">
<title>
<![CDATA[
Dual-view light-sheet imaging through tilted glass interface using a deformable mirror 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.10.20.345306v1?rss=1"
</link>
<description><![CDATA[
Light-sheet microscopy has become one of the primary tools for imaging live developing organisms because of its high speed, low phototoxicity, and optical sectioning capabilities. Detection from multiple sides (multi-view imaging) additionally allows nearly isotropic resolution via computational merging of the views. However, conventional light-sheet microscopes require that the sample is suspended in a gel to allow optical access from two or more sides. At the same time, the use of microfluidic devices is highly desirable for many experiments, but geometric constrains and strong optical aberrations caused by the coverslip titled relative to objectives make the use of multi-view lightsheet challenging for microfluidics.

In this paper we describe the use of adaptive optics (AO) to enable multi-view light-sheet microscopy in such microfluidic setup by correcting optical aberrations introduced by the tilted coverslip. The optimal shape of deformable mirror is computed by an iterative stochastic gradient-descent algorithm that optimizes PSF in two orthogonal planes simultaneously. Simultaneous AO correction in two optical arms is achieved via a knife-edge mirror that splits excitation path and combines the detection path.

We characterize the performance of this novel microscope setup and, by dual-view light-sheet imaging of C.elegans inside a microfluidic channel, demonstrate a drastic improvement of image quality due to AO and dual-view reconstruction. Our microscope design allows multi-view light-sheet microscopy with microfluidic devices for precisely controlled experimental conditions and high-content screening.
]]></description>
<dc:creator>Vladimirov, N.</dc:creator>
<dc:creator>Preusser, F.</dc:creator>
<dc:creator>Wisniewski, J.</dc:creator>
<dc:creator>Yaniv, Z.</dc:creator>
<dc:creator>Desai, R. A.</dc:creator>
<dc:creator>Woehler, A.</dc:creator>
<dc:creator>Preibisch, S.</dc:creator>
<dc:date>2020-10-21</dc:date>
<dc:identifier>doi:10.1101/2020.10.20.345306</dc:identifier>
<dc:title><![CDATA[Dual-view light-sheet imaging through tilted glass interface using a deformable mirror]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.10.28.359240v1?rss=1">
<title>
<![CDATA[
Whole-genome analysis of Nigerian patients with breast cancer reveals ethnic-driven somatic evolution and distinct genomic subtypes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.10.28.359240v1?rss=1"
</link>
<description><![CDATA[
Black women of African ancestry experience more aggressive breast cancer with higher mortality rates than White women of European ancestry. Although inter-ethnic germline variation is known, differential somatic evolution has not been investigated in detail. Analysis of deep whole genomes of 97 breast tumors, with RNA-seq in a subset, from indigenous African patients in Nigeria in comparison to The Cancer Genome Atlas (n=76) revealed a higher rate of genomic instability and increased intra-tumoral heterogeneity as well as a unique genomic subtype defined by early clonal GATA3 mutations and a 10.5-year younger age at diagnosis. We also found evidence for non-coding mutations in two novel drivers (ZNF217 and SYPL1) and a novel INDEL signature strongly associated with African ancestry proportion. This comprehensive analysis of an understudied population underscores the need to incorporate diversity of genomes as a key parameter in fundamental research with potential to tailor clinical intervention and promote equity in precision oncology care.
]]></description>
<dc:creator>Ansari-Pour, N.</dc:creator>
<dc:creator>Zheng, Y.</dc:creator>
<dc:creator>Pitt, J. J.</dc:creator>
<dc:creator>Dentro, S.</dc:creator>
<dc:creator>Yoshimatsu, T. F.</dc:creator>
<dc:creator>Sanni, A.</dc:creator>
<dc:creator>Ajani, M.</dc:creator>
<dc:creator>Woodard, A.</dc:creator>
<dc:creator>Rajagopal, P. S.</dc:creator>
<dc:creator>Fitzgerald, D.</dc:creator>
<dc:creator>Gruber, A. J.</dc:creator>
<dc:creator>Odetunde, A.</dc:creator>
<dc:creator>Popoola, A.</dc:creator>
<dc:creator>Falusi, A. G.</dc:creator>
<dc:creator>Babalola, C. P.</dc:creator>
<dc:creator>Ogundiran, T.</dc:creator>
<dc:creator>Obafunwa, J.</dc:creator>
<dc:creator>Ojengbede, O.</dc:creator>
<dc:creator>Ibrahim, N.</dc:creator>
<dc:creator>Barretina, J.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>Chen, M.</dc:creator>
<dc:creator>White, K. P.</dc:creator>
<dc:creator>Huo, D.</dc:creator>
<dc:creator>Wedge, D. C.</dc:creator>
<dc:creator>Olopade, O. I.</dc:creator>
<dc:date>2020-10-28</dc:date>
<dc:identifier>doi:10.1101/2020.10.28.359240</dc:identifier>
<dc:title><![CDATA[Whole-genome analysis of Nigerian patients with breast cancer reveals ethnic-driven somatic evolution and distinct genomic subtypes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.10.28.360123v1?rss=1">
<title>
<![CDATA[
GDV1 C-terminal truncation of 39 amino acids disrupts sexual commitment in Plasmodium falciparum 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.10.28.360123v1?rss=1"
</link>
<description><![CDATA[
Malaria is a mosquito-borne disease caused by apicomplexan parasites of the genus Plasmodium. Completion of the parasites life cycle depends on the transmission of sexual stages, the gametocytes, from an infected human host to the mosquito vector. Sexual commitment occurs in only a small fraction of asexual blood stage parasites and is initiated by external cues. The gametocyte development protein 1 (GDV1) has been described as a key facilitator to trigger sexual commitment. GDV1 interacts with the silencing factor heterochromatin protein 1 (HP1), leading to its dissociation from heterochromatic DNA at the genomic locus encoding AP2-G, the master transcription factor of gametocytogenesis. How this process is regulated is not known. In this study we have addressed the role of protein kinases implicated in gametocyte development. From a pool of available protein kinase KO lines, we identified two kinase knockout lines which fail to produce gametocytes. However, independent genetic verification revealed that both kinases are not required for gametocytogenesis but both lines harbour the same mutation that leads to a truncation in the extreme C-terminus of GDV1. Introduction of the identified nonsense mutation into the genome of wild type parasite lines replicates the observed phenotype. Using a GDV1 overexpression line we show that the truncation in the GDV1 C-terminus does neither interfere with the nuclear import of GDV1 nor its interaction with HP1 in vitro, but appears important to sustain GDV1 protein levels and thereby sexual commitment.

ImportanceTransmission of malaria causing Plasmodium species by mosquitos requires the parasite to change from a continuously growing asexual parasite form growing in the blood, to a sexually differentiated form, the gametocyte. Only a small subset of asexual parasites differentiates into gametocytes that are taken up by the mosquito. Transmission represents a bottleneck in the lifecycle of the parasite, so a molecular understanding of the events that lead to stage conversion may identify novel intervention points. Here we screened a subset of kinases we hypothesized to play a role in this process. While we did not identify kinases required for sexual conversion, we identified a mutation in the C-terminus of the Gametocyte Development 1 protein (GDV1), which abrogates sexual development. The mutation destabilises the protein but not its interaction with its cognate binding partner HP1. This suggest an important role for the GDV1 C-terminus beyond trafficking and protein stability.
]]></description>
<dc:creator>Tiburcio, M.</dc:creator>
<dc:creator>Hitz, E.</dc:creator>
<dc:creator>Niederwieser, I.</dc:creator>
<dc:creator>Kelly, G.</dc:creator>
<dc:creator>Davies, H.</dc:creator>
<dc:creator>Doerig, C.</dc:creator>
<dc:creator>Billker, O.</dc:creator>
<dc:creator>Voss, T. S.</dc:creator>
<dc:creator>Treeck, M.</dc:creator>
<dc:date>2020-10-30</dc:date>
<dc:identifier>doi:10.1101/2020.10.28.360123</dc:identifier>
<dc:title><![CDATA[GDV1 C-terminal truncation of 39 amino acids disrupts sexual commitment in Plasmodium falciparum]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.03.366252v1?rss=1">
<title>
<![CDATA[
Copy number-aware deconvolution of tumor-normal DNA methylation profiles 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.03.366252v1?rss=1"
</link>
<description><![CDATA[
Aberrant methylation is a hallmark of cancer, but bulk tumor data is confounded by admixed normal cells and copy number changes. Here, we introduce Copy number-Aware Methylation Deconvolution Analysis of Cancers (CAMDAC; https://github.com/VanLoo-lab/CAMDAC), which outputs tumor purity, allele-specific copy number and deconvolved methylation estimates. We apply CAMDAC to 122 multi-region samples from 38 TRACERx non-small cell lung cancers profiled by reduced representation bisulfite sequencing. CAMDAC copy number profiles parallel those derived from genome sequencing and highlight widespread chromosomal instability. Deconvolved polymorphism-independent methylation rates enable unbiased tumor-normal and tumor-tumor differential methylation calling. Read-phasing validates CAMDAC methylation rates and directly links genotype and epitype. We show increased epigenetic instability in adenocarcinoma vs. squamous cell carcinoma, frequent hypermethylation at sites carrying somatic mutations, and parallel copy number losses and methylation changes at imprinted loci. Unlike bulk methylomes, CAMDAC profiles recapitulate tumor phylogenies and evidence distinct patterns of epigenetic heterogeneity in lung cancer.
]]></description>
<dc:creator>Larose Cadieux, E.</dc:creator>
<dc:creator>Tanic, M.</dc:creator>
<dc:creator>Wilson, G. A.</dc:creator>
<dc:creator>Baker, T.</dc:creator>
<dc:creator>Dietzen, M.</dc:creator>
<dc:creator>Dhami, P.</dc:creator>
<dc:creator>Vaikkinen, H.</dc:creator>
<dc:creator>Watkins, T. B. K.</dc:creator>
<dc:creator>Kanu, N.</dc:creator>
<dc:creator>Veeriah, S.</dc:creator>
<dc:creator>Jamal-Hanjani, M.</dc:creator>
<dc:creator>McGranahan, N.</dc:creator>
<dc:creator>Feber, A.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>TRACERx Consortium,</dc:creator>
<dc:creator>Beck, S.</dc:creator>
<dc:creator>Demeulemeester, J.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:date>2020-11-04</dc:date>
<dc:identifier>doi:10.1101/2020.11.03.366252</dc:identifier>
<dc:title><![CDATA[Copy number-aware deconvolution of tumor-normal DNA methylation profiles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.03.367433v1?rss=1">
<title>
<![CDATA[
9p21 Loss Defines the Evolutionary Patterns of Aggressive Renal Cell Carcinomas 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.03.367433v1?rss=1"
</link>
<description><![CDATA[
Dedifferentiation and acquisition of chromosomal instability in renal cell carcinoma portends dismal prognosis and aggressive clinical behavior. However, the absence of reliable experimental models dramatically impacts the understanding of mechanisms underlying malignant progression. Here we established an in vivo genetic platform to rapidly generate somatic mosaic genetically engineerd immune-competent mouse models of renal tumors, recapitulating the genomic and phenotypic features of these malignancies. Leveraging somatic chromosomal engineering, we demonstrated that ablation of the murine locus syntenic to human 9p21 drives the rapid expansion of aggressive mesenchymal clones with prominent metastatic behavior, characterized by early emergence of chromosomal instability, whole-genome duplication, and conserved patterns of aneuploidy. This model of punctuated equilibrium provides a remarkable example of cross-species convergent evolution.

SignificanceTo better understand the role of 9p21 in malignant progression, we generated a somatic mosaic GEMM of renal cancer, capturing the histological, genomic and evolutionary features of human disease. With this technology we demonstrated a critica role of 9p21 loss in metastatic evolution of RCC and provide a unique tool for testing new therapeutic treatments.
]]></description>
<dc:creator>Carbone, F.</dc:creator>
<dc:creator>Huang, J. K.</dc:creator>
<dc:creator>Perelli, L.</dc:creator>
<dc:creator>Del Poggetto, E.</dc:creator>
<dc:creator>Gutschner, T.</dc:creator>
<dc:creator>Tomihara, H.</dc:creator>
<dc:creator>Soeung, M.</dc:creator>
<dc:creator>Lam, T. N. A.</dc:creator>
<dc:creator>Xia, R.</dc:creator>
<dc:creator>Zhu, C.</dc:creator>
<dc:creator>Song, X.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Sircar, K.</dc:creator>
<dc:creator>Malouf, G. G.</dc:creator>
<dc:creator>Sgambato, A.</dc:creator>
<dc:creator>Karam, J. A.</dc:creator>
<dc:creator>Gao, J.</dc:creator>
<dc:creator>Jonasch, E.</dc:creator>
<dc:creator>Viale, A.</dc:creator>
<dc:creator>Draetta, G. F.</dc:creator>
<dc:creator>Futreal, A.</dc:creator>
<dc:creator>Bakouny, Z.</dc:creator>
<dc:creator>Van Allen, E. M.</dc:creator>
<dc:creator>Choueiri, T.</dc:creator>
<dc:creator>Msaouel, P.</dc:creator>
<dc:creator>Litchfield, K.</dc:creator>
<dc:creator>Turajlic, S.</dc:creator>
<dc:creator>Heffernan, T. P.</dc:creator>
<dc:creator>Chen, Y. B.</dc:creator>
<dc:creator>DiNatale, R. G.</dc:creator>
<dc:creator>Hakimi, A. A.</dc:creator>
<dc:creator>Bristow, C. A.</dc:creator>
<dc:creator>Tannir, N. M.</dc:creator>
<dc:creator>Carugo, A.</dc:creator>
<dc:creator>Genovese, G.</dc:creator>
<dc:date>2020-11-04</dc:date>
<dc:identifier>doi:10.1101/2020.11.03.367433</dc:identifier>
<dc:title><![CDATA[9p21 Loss Defines the Evolutionary Patterns of Aggressive Renal Cell Carcinomas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.05.369629v1?rss=1">
<title>
<![CDATA[
Single-molecule analysis reveals cooperative stimulation of Rad51 filament nucleation and growth by mediator proteins 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.05.369629v1?rss=1"
</link>
<description><![CDATA[
Homologous recombination (HR) is an essential DNA double-strand break (DSBs) repair mechanism frequently inactivated in cancer. During HR, RAD51 forms nucleoprotein filaments on RPA-coated resected DNA and catalyses strand invasion into homologous duplex DNA. How RAD51 displaces RPA and assembles into long HR-proficient filaments remains uncertain. Here, we employ single-molecule imaging to investigate the mechanism of nematode RAD-51 filament growth in the presence of BRC-2 (BRCA2) and RAD-51 paralogs, RFS-1/RIP-1. BRC-2 nucleates RAD-51 on RPA-coated DNA, while RFS-1/RIP-1 acts as a  chaperone to promote 3 to 5 filament growth via highly dynamic engagement with 5 filament ends. Inhibiting ATPase or mutation in RFS-1 Walker box leads to RFS-1/RIP-1 retention on RAD-51 filaments and hinders growth. rfs-1 Walker box mutants display sensitivity to DNA damage and accumulate RAD-51 complexes non-functional for HR in vivo. Our work reveals the mechanism of RAD-51 nucleation and filament growth in the presence of recombination mediators.
]]></description>
<dc:creator>Belan, O.</dc:creator>
<dc:creator>Barroso, C.</dc:creator>
<dc:creator>Kaczmarczyk, A.</dc:creator>
<dc:creator>Anand, R.</dc:creator>
<dc:creator>Federico, S.</dc:creator>
<dc:creator>O'Reilly, N.</dc:creator>
<dc:creator>Newton, M. D.</dc:creator>
<dc:creator>Maeots, E.</dc:creator>
<dc:creator>Enchev, R. I.</dc:creator>
<dc:creator>Martinez-Perez, E.</dc:creator>
<dc:creator>Rueda, D. S.</dc:creator>
<dc:creator>Boulton, S. J.</dc:creator>
<dc:date>2020-11-05</dc:date>
<dc:identifier>doi:10.1101/2020.11.05.369629</dc:identifier>
<dc:title><![CDATA[Single-molecule analysis reveals cooperative stimulation of Rad51 filament nucleation and growth by mediator proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.10.376491v1?rss=1">
<title>
<![CDATA[
Temporal patterning of the central nervous system by a shared transcription factor code 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.10.376491v1?rss=1"
</link>
<description><![CDATA[
The molecular mechanisms that ensure the reproducible generation of neuronal diversity in the vertebrate nervous system are incompletely understood. Here we provide evidence of a temporal patterning program consisting of cohorts of transcription factors expressed in neurons generated at successive developmental timepoints. This program acts in parallel to spatial patterning, diversifying neurons throughout the nervous system and in neurons differentiated in-vitro from stem cells. We demonstrate the TGF{beta} signalling pathway controls the pace of the temporal program. Furthermore, targeted perturbation of components of the temporal program, Nfia and Nfib, reveals their requirement for the generation of late-born neuronal subtypes. Together, our results provide evidence for the existence of a previously unappreciated global temporal program of neuronal subtype identity and suggest that the integration of spatial and temporal patterning programs diversifies and organises neuronal subtypes in the vertebrate nervous system.
]]></description>
<dc:creator>Sagner, A.</dc:creator>
<dc:creator>Zhang, I.</dc:creator>
<dc:creator>Watson, T.</dc:creator>
<dc:creator>Lazaro, J.</dc:creator>
<dc:creator>Melchionda, M.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:date>2020-11-10</dc:date>
<dc:identifier>doi:10.1101/2020.11.10.376491</dc:identifier>
<dc:title><![CDATA[Temporal patterning of the central nervous system by a shared transcription factor code]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.10.376129v1?rss=1">
<title>
<![CDATA[
ECM remodeling and spatial cell cycle coordination determine tissue growth kinetics 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.10.376129v1?rss=1"
</link>
<description><![CDATA[
During development, multicellular organisms undergo stereotypical patterns of tissue growth to yield organs of highly reproducible sizes and shapes. How this process is orchestrated remains unclear. Analysis of the temporal dynamics of tissue growth in the Drosophila abdomen reveals that cell cycle times are spatially correlated and that growth termination occurs through the rapid emergence of a population of arrested cells rather than a gradual slowing down of cell cycle time. Reduction in apical tension associated with tissue crowding has been proposed as a developmental growth termination mechanism. Surprisingly, we find that growth arrest in the abdomen occurs while apical tension increases, showing that in this tissue a reduction in tension does not underlie the mechanism of growth arrest. However, remodeling of the extracellular matrix is necessary for tissue expansion. Thus, changes in the tissue microenvironment, and a rapid exit from proliferation, control the formation of the adult Drosophila abdomen.
]]></description>
<dc:creator>Ainslie, A. P.</dc:creator>
<dc:creator>Davis, J. R.</dc:creator>
<dc:creator>Williamson, J. J.</dc:creator>
<dc:creator>Ferreira, A.</dc:creator>
<dc:creator>Torres-Sanchez, A.</dc:creator>
<dc:creator>Hoppe, A.</dc:creator>
<dc:creator>Mangione, F.</dc:creator>
<dc:creator>Smith, M. B.</dc:creator>
<dc:creator>Martin-Blanco, E.</dc:creator>
<dc:creator>Salbreux, G.</dc:creator>
<dc:creator>Tapon, N.</dc:creator>
<dc:date>2020-11-10</dc:date>
<dc:identifier>doi:10.1101/2020.11.10.376129</dc:identifier>
<dc:title><![CDATA[ECM remodeling and spatial cell cycle coordination determine tissue growth kinetics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.11.378869v1?rss=1">
<title>
<![CDATA[
A FGF2-mediated incoherent feedforward loop inducesErk inhibition and promotes naive pluripotency 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.11.378869v1?rss=1"
</link>
<description><![CDATA[
Naive pluripotency is a transient state during mammalian development that can be recapitulated indefinitely in vitro by inhibition of the mitogen-activated protein kinase (MAPK/Erk) signalling and activation of STAT and Wnt pathways. How Erk is inhibited in vivo to promote naive pluripotency remains largely unknown. By combining live cell imaging and quantitative proteomics we found that FGF2, a known Erk activator and pro-differentiation cue, induces instead long-term Erk inhibition in both ES cells and mouse embryos. We show that Erk inhibition results from a FGF2-induced incoherent feedforward loop. Importantly, we see that FGF2 induces up-regulation of naive pluripotency factors, down-regulation of DNA methylation by suppression of de novo DNA methylases thereby helping maintain naive pluripotency. We show that FGF2 is expressed maternally and propose that integration of signals from the embryos niche may contribute to the generation of embryonic lineages with the right cell proportions. We suggest that feedforward regulation may play a role driving transient, reversible developmental transitions.
]]></description>
<dc:creator>Gharibi, B.</dc:creator>
<dc:creator>Goncalves, E.</dc:creator>
<dc:creator>Nashun, B.</dc:creator>
<dc:creator>Montoya, A.</dc:creator>
<dc:creator>Mankalow, K.</dc:creator>
<dc:creator>Strohbuecker, S.</dc:creator>
<dc:creator>Malik Sheriff, R. S.</dc:creator>
<dc:creator>Cicarrelli, A.</dc:creator>
<dc:creator>Carvalho, J.</dc:creator>
<dc:creator>Nye, E.</dc:creator>
<dc:creator>Kramer, H.</dc:creator>
<dc:creator>Rosewell, I.</dc:creator>
<dc:creator>Hajkova, P.</dc:creator>
<dc:creator>Beltrao, P.</dc:creator>
<dc:creator>Santos, S.</dc:creator>
<dc:date>2020-11-11</dc:date>
<dc:identifier>doi:10.1101/2020.11.11.378869</dc:identifier>
<dc:title><![CDATA[A FGF2-mediated incoherent feedforward loop inducesErk inhibition and promotes naive pluripotency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.14.381277v1?rss=1">
<title>
<![CDATA[
Can a nick promote partial genome re-replication? 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.14.381277v1?rss=1"
</link>
<description><![CDATA[
Single-stranded DNA breaks, including simple nicks, are amongst the most common forms of DNA damage in cells. They can be readily repaired by ligation; however, if a nick occurs just ahead of an approaching replisome, the outcome is a  collapsed replication fork in which the nick is converted into a single-ended double-strand DNA break. Attention has largely focused on the processes by which this broken end is used to prime replication restart. We realized that in eukaryotic cells, where replication initiates from multiple replication origins, a second fork converging on the collapsed fork offers additional opportunities for repair, but also generates a substrate that can promote localized re-replication. We have modelled this with purified proteins in vitro and have found that there is, indeed, an additional hazard that eukaryotic replisomes face. We discuss how this problem might be mitigated.
]]></description>
<dc:creator>Johansson, E.</dc:creator>
<dc:creator>Diffley, J. F. X.</dc:creator>
<dc:date>2020-11-15</dc:date>
<dc:identifier>doi:10.1101/2020.11.14.381277</dc:identifier>
<dc:title><![CDATA[Can a nick promote partial genome re-replication?]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.10.376186v1?rss=1">
<title>
<![CDATA[
EphB6 regulates TFEB-lysosomal pathway and survival of disseminated indolent breast cancer cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.10.376186v1?rss=1"
</link>
<description><![CDATA[
Late relapse of disseminated cancer cells is a common feature of some types of tumors. Several intrinsic and extrinsic factors have been shown to affect reawakening of disseminated dormant cancer cells (DDCCs); however, the signals and processes sustaining survival of DDCCs in a foreign environment are still poorly understood. We have recently shown that crosstalk with lung epithelial cells promotes persistence of DDCCs from estrogen receptor positive (ER+) breast tumors. Here we show that TFEB-lysosomal axis is activated in DDCCs and that it is modulated by the pro-survival ephrin receptor EphB6. TFEB lysosomal direct targets are enriched in DDCCs in vivo and correlate with relapse in ER+ breast cancer patients. Direct contact of DDCCs with alveolar type I-like lung epithelial cells drives lysosomal accumulation and EphB6 induction. EphB6 contributes to TFEB transcriptional activity and lysosome formation in DDCCs in vitro and in vivo, and supports survival of DDCCs in coculture and in vivo. Furthermore, signaling from EphB6 promotes the proliferative response of surrounding lung parenchymal cells in vivo.
]]></description>
<dc:creator>MONTAGNER, M.</dc:creator>
<dc:creator>Zangrossi, M.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Romani, P.</dc:creator>
<dc:creator>Ratcliffe, C. D. H.</dc:creator>
<dc:creator>Hooper, S.</dc:creator>
<dc:creator>Dori, M.</dc:creator>
<dc:creator>Forcato, M.</dc:creator>
<dc:creator>Bicciato, S.</dc:creator>
<dc:creator>Dupont, S.</dc:creator>
<dc:creator>Sahai, E.</dc:creator>
<dc:date>2020-11-10</dc:date>
<dc:identifier>doi:10.1101/2020.11.10.376186</dc:identifier>
<dc:title><![CDATA[EphB6 regulates TFEB-lysosomal pathway and survival of disseminated indolent breast cancer cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.17.377705v1?rss=1">
<title>
<![CDATA[
USP28 deletion and small molecule inhibition destabilises c-Myc and elicitsregression of squamous cell lung carcinoma 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.17.377705v1?rss=1"
</link>
<description><![CDATA[
Lung squamous cell carcinoma (LSCC) is a considerable global health burden, with an incidence of over 600,000 cases per year. Treatment options are limited, and patient 5-year survival rate is less than 5%. The ubiquitin specific protease 28 (USP28) has been implicated in tumorigenesis through its stabilization of the oncoprotein c-MYC. Here, we show that genetic inactivation of Usp28 induced regression of established murine LSCC lung tumors. We developed a small molecule that inhibits USP28 activity in the low nanomole range. While displaying cross-reactivity against the closest homologue USP25, this inhibitor showed a high degree of selectivity over other deubiquitinases. USP28 inhibitor treatment resulted in a dramatic decrease in c-Myc proteins levels and consequently induced substantial regression of autochthonous murine LSCC tumors and human LSCC xenografts, thereby phenocopying the effect observed by genetic deletion. Thus, USP28 may represent a promising therapeutic target for the treatment of squamous cell lung carcinoma.
]]></description>
<dc:creator>Ruiz, E. J.</dc:creator>
<dc:creator>Pinto-Fernandez, A.</dc:creator>
<dc:creator>Turnbull, A. P.</dc:creator>
<dc:creator>Lan, L.</dc:creator>
<dc:creator>Charlton, T. M.</dc:creator>
<dc:creator>Scott, H. C.</dc:creator>
<dc:creator>Damianou, A.</dc:creator>
<dc:creator>Vere, G.</dc:creator>
<dc:creator>Riising, E. M.</dc:creator>
<dc:creator>Da Costa, C.</dc:creator>
<dc:creator>Krajewski, W. W.</dc:creator>
<dc:creator>Guerin, D.</dc:creator>
<dc:creator>Kearns, J.</dc:creator>
<dc:creator>Ioannidis, S.</dc:creator>
<dc:creator>Katz, M.</dc:creator>
<dc:creator>O'Connell, J. C.</dc:creator>
<dc:creator>Moncaut, N.</dc:creator>
<dc:creator>Rosewell, I.</dc:creator>
<dc:creator>Nye, E.</dc:creator>
<dc:creator>Jones, N.</dc:creator>
<dc:creator>Heride, C.</dc:creator>
<dc:creator>Gersch, M.</dc:creator>
<dc:creator>Wu, M.</dc:creator>
<dc:creator>Dinsmore, C. J.</dc:creator>
<dc:creator>Hammonds, T. R.</dc:creator>
<dc:creator>Kim, S.</dc:creator>
<dc:creator>Komander, D.</dc:creator>
<dc:creator>Urbe, S.</dc:creator>
<dc:creator>Clague, M. J.</dc:creator>
<dc:creator>Kessler, B. M.</dc:creator>
<dc:creator>Behrens, A.</dc:creator>
<dc:date>2020-11-17</dc:date>
<dc:identifier>doi:10.1101/2020.11.17.377705</dc:identifier>
<dc:title><![CDATA[USP28 deletion and small molecule inhibition destabilises c-Myc and elicitsregression of squamous cell lung carcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.18.388306v1?rss=1">
<title>
<![CDATA[
Subtractive CRISPR screen identifies factors involved in non-canonical LC3 lipidation. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.18.388306v1?rss=1"
</link>
<description><![CDATA[
Although commonly associated with autophagosomes, LC3 can also be recruited to membranes in a variety of non-canonical contexts. These include responses to ionophores such as the M2 proton channel of influenza A virus. LC3 is attached to membranes by covalent lipidation that depends on recruitment of the ATG5-12-16L1 complex. Non-canonical LC3 lipidation requires the C-terminal WD40 domain of ATG16L1 that is dispensable for canonical autophagy. We devised a subtractive CRISPR knock-out screening strategy to investigate the requirements for non-canonical LC3-lipidation. This correctly identified the enzyme complexes directly responsible for LC3-lipidation. We additionally identified the RALGAP complex as important for M2-induced, but not ionophore drug induced LC3 lipidation. In contrast, we identified ATG4D as responsible for LC3 recycling in M2-induced and basal LC3-lipidation. Identification of a vacuolar ATPase subunit in the screen suggested a common mechanism for non-canonical LC3 recruitment. Influenza-induced and ionophore drug induced LC3-lipidation leads to association of the vacuolar ATPase and ATG16L1 and can be antagonised by Salmonella SopF. LC3 recruitment to erroneously neutral compartments may therefore represent a response to damage caused by diverse invasive pathogens.
]]></description>
<dc:creator>Ulferts, R.</dc:creator>
<dc:creator>Marcassa, E.</dc:creator>
<dc:creator>Timimi, L.</dc:creator>
<dc:creator>Lee, L. C.</dc:creator>
<dc:creator>Daley, A.</dc:creator>
<dc:creator>Montaner, B.</dc:creator>
<dc:creator>Turner, S. D.</dc:creator>
<dc:creator>Florey, O.</dc:creator>
<dc:creator>Baillie, J. K.</dc:creator>
<dc:creator>Beale, R.</dc:creator>
<dc:date>2020-11-18</dc:date>
<dc:identifier>doi:10.1101/2020.11.18.388306</dc:identifier>
<dc:title><![CDATA[Subtractive CRISPR screen identifies factors involved in non-canonical LC3 lipidation.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.19.390146v1?rss=1">
<title>
<![CDATA[
Adipose Triglyceride Lipase protects the endocytosis of renal cells on a high fat diet in Drosophila 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.19.390146v1?rss=1"
</link>
<description><![CDATA[
Obesity-related renal lipotoxicity and chronic kidney disease (CKD) are prevalent pathologies with complex aetiologies. One hallmark of renal lipotoxicity is the ectopic accumulation of lipid droplets in kidney podocytes and in proximal tubule cells. Renal lipid droplets are observed in human CKD patients and in high-fat diet rodent models but their precise role remains unclear. Here, we establish a high-fat diet model in Drosophila that recapitulates renal lipid droplets and several other aspects of mammalian CKD. Cell-type specific genetic manipulations show that lipid can overflow from adipose tissue and is taken up by renal cells called nephrocytes. A high-fat diet drives nephrocyte lipid uptake via the multiligand receptor Cubilin, leading to the ectopic accumulation of lipid droplets. These nephrocyte lipid droplets correlate with ER and mitochondrial deficits, as well as with impaired macromolecular endocytosis, a key conserved function of renal cells. Nephrocyte knockdown of diglyceride acyltransferase 1 (DGAT1), overexpression of adipose triglyceride lipase (ATGL) and epistasis tests together reveal that fatty acid flux through the lipid droplet triglyceride compartment protects the ER, mitochondria and endocytosis of renal cells. Strikingly, boosting nephrocyte expression of the lipid droplet resident enzyme ATGL is sufficient to rescue high-fat diet induced defects in renal endocytosis. Moreover, endocytic rescue requires a conserved mitochondrial regulator, peroxisome proliferator-activated receptor-gamma coactivator 1 (PGC1). This study demonstrates that lipid droplet lipolysis counteracts the harmful effects of a high-fat diet via a mitochondrial pathway that protects renal endocytosis. It also provides a genetic strategy for determining whether lipid droplets in different biological contexts function primarily to release beneficial or to sequester toxic lipids.
]]></description>
<dc:creator>Lubojemska, A.</dc:creator>
<dc:creator>Stefana, M. I.</dc:creator>
<dc:creator>Lampe, L.</dc:creator>
<dc:creator>Yoshimura, A.</dc:creator>
<dc:creator>Burrell, A.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:creator>Gould, A. P.</dc:creator>
<dc:date>2020-11-20</dc:date>
<dc:identifier>doi:10.1101/2020.11.19.390146</dc:identifier>
<dc:title><![CDATA[Adipose Triglyceride Lipase protects the endocytosis of renal cells on a high fat diet in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.20.391896v1?rss=1">
<title>
<![CDATA[
Highly multiplexed spatially resolved gene expression profiling of mouse organogenesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.20.391896v1?rss=1"
</link>
<description><![CDATA[
Transcriptional and epigenetic profiling of single-cells has advanced our knowledge of the molecular bases of gastrulation and early organogenesis. However, current approaches rely on dissociating cells from tissues, thereby losing the crucial spatial context that is necessary for understanding cell and tissue interactions during development. Here, we apply an image-based single-cell transcriptomics method, seqFISH, to simultaneously and precisely detect mRNA molecules for 387 selected target genes in 8-12 somite stage mouse embryo tissue sections. By integrating spatial context and highly multiplexed transcriptional measurements with two single-cell transcriptome atlases we accurately characterize cell types across the embryo and demonstrate how spatially-resolved expression of genes not profiled by seqFISH can be imputed. We use this high-resolution spatial map to characterize fundamental steps in the patterning of the midbrain-hindbrain boundary and the developing gut tube. Our spatial atlas uncovers axes of resolution that are not apparent from single-cell RNA sequencing data - for example, in the gut tube we observe early dorsal-ventral separation of esophageal and tracheal progenitor populations. In sum, by computationally integrating high-resolution spatially-resolved gene expression maps with single-cell genomics data, we provide a powerful new approach for studying how and when cell fate decisions are made during early mammalian development.
]]></description>
<dc:creator>Lohoff, T.</dc:creator>
<dc:creator>Ghazanfar, S.</dc:creator>
<dc:creator>Missarova, A.</dc:creator>
<dc:creator>Koulena, N.</dc:creator>
<dc:creator>Pierson, N.</dc:creator>
<dc:creator>Griffiths, J. A.</dc:creator>
<dc:creator>Bardot, E. S.</dc:creator>
<dc:creator>Eng, C.-H. L.</dc:creator>
<dc:creator>Tyser, R. C. V.</dc:creator>
<dc:creator>Argelaguet, R.</dc:creator>
<dc:creator>Guibentif, C.</dc:creator>
<dc:creator>Srinivas, S.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Simons, B. D.</dc:creator>
<dc:creator>Hadjantonakis, A.-K.</dc:creator>
<dc:creator>Gottgens, B.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:creator>Nichols, J.</dc:creator>
<dc:creator>Cai, L.</dc:creator>
<dc:creator>Marioni, J. C.</dc:creator>
<dc:date>2020-11-21</dc:date>
<dc:identifier>doi:10.1101/2020.11.20.391896</dc:identifier>
<dc:title><![CDATA[Highly multiplexed spatially resolved gene expression profiling of mouse organogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.20.391300v1?rss=1">
<title>
<![CDATA[
2019 Association of Biomolecular Resource Facilities Multi-Laboratory Data-Independent Acquisition Study 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.20.391300v1?rss=1"
</link>
<description><![CDATA[
Despite the advantages of fewer missing values by collecting fragment ion data on all analytes in the sample, as well as the potential for deeper coverage, the adoption of data-independent acquisition (DIA) in core facility settings has been slow. The Association of Biomolecular Resource Facilities conducted a large interlaboratory study to evaluate DIA performance in laboratories with various instrumentation. Participants were supplied with generic methods and a uniform set of test samples. The resulting 49 DIA datasets act as benchmarks and have utility in education and tool development. The sample set consisted of a tryptic HeLa digest spiked with high or low levels of four exogenous proteins. Data are available in MassIVE MSV000086479. Additionally, we demonstrate how the data can be analysed by focusing on two datasets using different library approaches and show the utility of select summary statistics. These data can be used by DIA newcomers, software developers, or DIA experts evaluating performance with different platforms, acquisition settings and skill levels.
]]></description>
<dc:creator>Neely, B. A.</dc:creator>
<dc:creator>Stemmer, P.</dc:creator>
<dc:creator>Searle, B. C.</dc:creator>
<dc:creator>Herring, L. E.</dc:creator>
<dc:creator>Martin, L.</dc:creator>
<dc:creator>Midha, M. K.</dc:creator>
<dc:creator>Phinney, B. S.</dc:creator>
<dc:creator>Shan, B.</dc:creator>
<dc:creator>Palmblad, M.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Jagtap, P. D.</dc:creator>
<dc:creator>Kirkpatrick, J. M.</dc:creator>
<dc:date>2020-11-20</dc:date>
<dc:identifier>doi:10.1101/2020.11.20.391300</dc:identifier>
<dc:title><![CDATA[2019 Association of Biomolecular Resource Facilities Multi-Laboratory Data-Independent Acquisition Study]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.25.397943v1?rss=1">
<title>
<![CDATA[
Synergistic CDK control pathways maintain cell size homeostasis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.25.397943v1?rss=1"
</link>
<description><![CDATA[
To coordinate cell size with cell division, cell size must be computed by the cyclin-CDK control network to trigger division appropriately. Here we dissect determinants of cyclin-CDK activity using a novel high-throughput single-cell in vivo system. We show that inhibitory phosphorylation of CDK encodes cell size information and works synergistically with PP2A to prevent division in smaller cells. However, even in the absence of all canonical regulators of cyclin-CDK, small cells with high cyclin-CDK levels are restricted from dividing. We find that diploid cells of equivalent size to haploid cells exhibit lower CDK activity in response to equal cyclin-CDK enzyme concentrations, suggesting that CDK activity is reduced by DNA concentration. Thus, multiple pathways directly regulate cyclin-CDK activity to maintain robust cell size homeostasis.
]]></description>
<dc:creator>Patterson, J. O.</dc:creator>
<dc:creator>Basu, S.</dc:creator>
<dc:creator>Rees, P.</dc:creator>
<dc:creator>Nurse, P.</dc:creator>
<dc:date>2020-11-25</dc:date>
<dc:identifier>doi:10.1101/2020.11.25.397943</dc:identifier>
<dc:title><![CDATA[Synergistic CDK control pathways maintain cell size homeostasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.25.398636v1?rss=1">
<title>
<![CDATA[
Intracellular localisation of Mycobacterium tuberculosis affects antibiotic efficacy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.25.398636v1?rss=1"
</link>
<description><![CDATA[
To be effective, chemotherapy against tuberculosis (TB) must kill the intracellular population of Mycobacterium tuberculosis (Mtb). However, how host cell environments affect antibiotic accumulation and efficacy remains elusive. Pyrazinamide (PZA) is a key antibiotic against TB, yet its behaviour is not fully understood. Here, by using correlative light, electron, and ion microscopy to image PZA at the subcellular level, we investigated how human macrophage environments affect PZA activity. We discovered that PZA accumulates heterogeneously between individual bacteria in multiple host cell environments. Crucially, Mtb phagosomal localisation and acidification increase PZA accumulation and efficacy. By imaging two antibiotics commonly used in combined TB therapy, we showed that bedaquiline (BDQ) significantly enhances PZA accumulation by a host cell mediated mechanism. Thus, intracellular localisation and specific microenvironments affect PZA accumulation and efficacy; explaining the potent in vivo efficacy compared to its modest in vitro activity and the critical contribution to TB combination chemotherapy.
]]></description>
<dc:creator>Santucci, P.</dc:creator>
<dc:creator>Greenwood, D.</dc:creator>
<dc:creator>Fearns, A.</dc:creator>
<dc:creator>Chen, K.</dc:creator>
<dc:creator>Jiang, H.</dc:creator>
<dc:creator>Gutierrez, M. G.</dc:creator>
<dc:date>2020-11-25</dc:date>
<dc:identifier>doi:10.1101/2020.11.25.398636</dc:identifier>
<dc:title><![CDATA[Intracellular localisation of Mycobacterium tuberculosis affects antibiotic efficacy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.29.402610v1?rss=1">
<title>
<![CDATA[
Coupling of mouse olfactory bulb projection neurons to fluctuating odour pulses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.29.402610v1?rss=1"
</link>
<description><![CDATA[
Odours are transported by turbulent air currents, creating complex temporal fluctuations in odour concentration. Recently, we have shown that mice can discriminate odour stimuli based on their temporal structure, indicating that information present in the temporal structure of odour plumes may be extracted by the mouse olfactory system. Here using in vivo electrophysiological recordings, we show that mitral and tufted cells (M/TCs), the projection neurons of the mouse olfactory bulb, can encode the dominant temporal frequencies present in odour stimuli up to frequencies of at least 20 Hz. We show that M/TCs couple their membrane potential to odour concentration fluctuations; coupling was variable between M/TCs but independent of the odour presented and with TCs displaying slightly elevated coupling compared to MCs in particular for higher frequency stimulation (20Hz). Pharmacologically blocking the inhibitory circuitry strongly modulated frequency coupling. Together this suggests that both cellular and circuit properties contribute to the encoding of temporal odour features in the mouse olfactory bulb.
]]></description>
<dc:creator>Dasgupta, D.</dc:creator>
<dc:creator>Warner, T. P. A.</dc:creator>
<dc:creator>Erskine, A.</dc:creator>
<dc:creator>Schaefer, A. T.</dc:creator>
<dc:date>2020-11-29</dc:date>
<dc:identifier>doi:10.1101/2020.11.29.402610</dc:identifier>
<dc:title><![CDATA[Coupling of mouse olfactory bulb projection neurons to fluctuating odour pulses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.12.01.393157v1?rss=1">
<title>
<![CDATA[
The metalloproteinase inhibitor Prinomastat reduces AML growth,prevents stem cell loss and improves chemotherapy effectiveness 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.01.393157v1?rss=1"
</link>
<description><![CDATA[
Acute myeloid leukemia (AML) is a blood cancer of the myeloid lineage. Its prognosis remains poor, highlighting the need for new therapeutic and precision medicine approaches. AML symptoms often include cytopenias, linked to loss of healthy hematopoietic stem and progenitor cells (HSPCs). The mechanism behind HSPC decline is complex and still poorly understood. Here, intravital microscopy (IVM) of a well-established experimental model of AML allows direct observation of the interactions between healthy and malignant cells in the bone marrow (BM), suggesting that physical dislodgment of healthy cells by AML through damaged vasculature may play an important role. Numerous human leukemia types, particularly MLL-AF9 samples, show high expression levels of multiple matrix metalloproteinases (MMPs). Therefore, we evaluate the therapeutic potential of the MMP inhibitor (MMPI) prinomastat. IVM analyses of treated mice reveal reduced vascular permeability and healthy cell clusters in circulation, and lower AML cell speed. Furthermore, treated mice have decreased BM infiltration, increased retention of healthy HSPCs in the BM and increased survival following chemotherapy. Overall, our results suggest that MMPIs could be a promising complementary therapy to reduce AML growth and limit the loss of HSPC and BM vascular damage caused by MLL-AF9 and possibly other AML subtypes.
]]></description>
<dc:creator>Pirillo, C.</dc:creator>
<dc:creator>Haltalli, M.</dc:creator>
<dc:creator>Gonzalez Anton, S.</dc:creator>
<dc:creator>Tini, V.</dc:creator>
<dc:creator>Kong, I.</dc:creator>
<dc:creator>Hawkins, E.</dc:creator>
<dc:creator>Falini, B.</dc:creator>
<dc:creator>Marra, A.</dc:creator>
<dc:creator>Duarte, D.</dc:creator>
<dc:creator>Lo Celso, C.</dc:creator>
<dc:date>2020-12-01</dc:date>
<dc:identifier>doi:10.1101/2020.12.01.393157</dc:identifier>
<dc:title><![CDATA[The metalloproteinase inhibitor Prinomastat reduces AML growth,prevents stem cell loss and improves chemotherapy effectiveness]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.30.405571v1?rss=1">
<title>
<![CDATA[
Molecular characterization of projection neuron subtypes in the mouse olfactory bulb 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.30.405571v1?rss=1"
</link>
<description><![CDATA[
Projection neurons (PNs) in the mammalian olfactory bulb (OB) receive direct input from the nose and project to diverse cortical and subcortical areas. Morphological and physiological studies have highlighted functional heterogeneity, yet no molecular markers have been described that delineate PN subtypes. Here, we used viral injections into olfactory cortex and fluorescent nucleus sorting to enrich PNs for high-throughput single nucleus and bulk RNA deep sequencing. Transcriptome analysis and RNA in situ hybridization identified three mitral and five tufted cell populations with characteristic transcription factor network topology and cell adhesion and excitability-related gene expression. Finally, by integrating bulk and snRNA-seq data we propose that different mitral cell populations selectively project to different regions of olfactory cortex. Together, we have identified potential molecular and gene regulatory mechanisms underlying PN diversity and provide new molecular entry points into studying the diverse functional roles of mitral and tufted cell subtypes.
]]></description>
<dc:creator>Zeppilli, S.</dc:creator>
<dc:creator>Ackels, T.</dc:creator>
<dc:creator>Attey, R.</dc:creator>
<dc:creator>Klimpert, N.</dc:creator>
<dc:creator>Boeing, S.</dc:creator>
<dc:creator>Crombach, A.</dc:creator>
<dc:creator>Schaefer, A.</dc:creator>
<dc:creator>Fleischmann, A.</dc:creator>
<dc:date>2020-12-02</dc:date>
<dc:identifier>doi:10.1101/2020.11.30.405571</dc:identifier>
<dc:title><![CDATA[Molecular characterization of projection neuron subtypes in the mouse olfactory bulb]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.12.04.411793v1?rss=1">
<title>
<![CDATA[
Recognition of Z-RNA by ADAR1 limits interferon responses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.04.411793v1?rss=1"
</link>
<description><![CDATA[
Nucleic acids are powerful triggers of innate immunity and can adopt the unusual Z-conformation. The p150 isoform of adenosine deaminase acting on RNA 1 (ADAR1) prevents aberrant interferon (IFN) induction and contains a Z-nucleic acid binding (Z) domain. We report that knock-in mice bearing two point mutations in the Z domain of ADAR1, which abolish binding to Z-form nucleic acids, spontaneously induced type I IFNs and IFN-stimulated genes (ISGs) in multiple organs. This included the lung where both stromal and haematopoietic cells displayed ISG induction in Adar1mZ/mZ mice. Concomitantly, Adar1mZ/mZ mice showed improved control of influenza A virus. The spontaneous IFN response in Adar1mZ/mZ mice required MAVS, implicating cytosolic RNA sensing. Finally, analysis of A-to-I changes revealed a specific requirement of ADAR1s Z domain in editing of a subset of RNAs. In summary, our results reveal that endogenous RNAs in Z-conformation have immunostimulatory potential that is curtailed by ADAR1.
]]></description>
<dc:creator>Tang, Q.</dc:creator>
<dc:creator>Rigby, R. E.</dc:creator>
<dc:creator>Young, G. R.</dc:creator>
<dc:creator>Korning-Hvidt, A.</dc:creator>
<dc:creator>Tang, T. K.</dc:creator>
<dc:creator>Bridgeman, A.</dc:creator>
<dc:creator>Townsend, A. R.</dc:creator>
<dc:creator>Kassiotis, G.</dc:creator>
<dc:creator>Rehwinkel, J.</dc:creator>
<dc:date>2020-12-04</dc:date>
<dc:identifier>doi:10.1101/2020.12.04.411793</dc:identifier>
<dc:title><![CDATA[Recognition of Z-RNA by ADAR1 limits interferon responses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.12.08.415828v1?rss=1">
<title>
<![CDATA[
Memory-like B cells emerging from germinal centres recycle through the subcapsular sinus 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.08.415828v1?rss=1"
</link>
<description><![CDATA[
Infection or vaccination leads to the development of germinal centers (GCs) where B cells evolve high affinity antigen receptors, eventually producing antibody-forming plasma cells or memory B cells. We followed the migratory pathways of B cells emerging from germinal centers (BEM) and found that many migrated into the lymph node subcapsular sinus (SCS) guided by sphingosine-1-phosphate (S1P). From there, B cells may exit the lymph node to enter distant tissues. Some BEM cells interacted with and took up antigen from SCS macrophages, followed by CCL21-guided return towards the GC. Disruption of local CCL21 gradients inhibited the recycling of BEM cells and resulted in less efficient adaption to antigenic variation. Our findings suggest that the recycling of BEM cells, that transport antigen and that contain the genetic code for B cell receptor variants, may support affinity maturation to antigenic drift.
]]></description>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Garcia-Ibanez, L.</dc:creator>
<dc:creator>Ulbricht, C.</dc:creator>
<dc:creator>Lok, L. S. C.</dc:creator>
<dc:creator>Dennison, T. W.</dc:creator>
<dc:creator>Ferdinand, J. R.</dc:creator>
<dc:creator>Mueller-Winkler, J.</dc:creator>
<dc:creator>Burnett, C. J. M.</dc:creator>
<dc:creator>Yam-Puc, J. C.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Brown, G.</dc:creator>
<dc:creator>Tybulewicz, V.</dc:creator>
<dc:creator>Rot, A.</dc:creator>
<dc:creator>Hauser, A. E.</dc:creator>
<dc:creator>Clatworthy, M. R.</dc:creator>
<dc:creator>Toellner, K.-M.</dc:creator>
<dc:date>2020-12-09</dc:date>
<dc:identifier>doi:10.1101/2020.12.08.415828</dc:identifier>
<dc:title><![CDATA[Memory-like B cells emerging from germinal centres recycle through the subcapsular sinus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.12.15.422831v1?rss=1">
<title>
<![CDATA[
Immunogenomic profile of colorectal cancer response to immune checkpoint blockade 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.15.422831v1?rss=1"
</link>
<description><![CDATA[
Colorectal cancers (CRCs) show variable response to immune checkpoint blockade, which can only partially be explained by the variability of tumour mutational burden. To dissect the cellular and molecular determinants of response we performed a multi-omic screen of 721 cancer regions from patients treated with Pembrolizumab (KEYNOTE 177 clinical trial) or Nivolumab. Multi-regional whole exome, RNA and T-cell receptor sequencing show that, within hypermutated CRCs, response to both anti-PD1 agents is not positively associated with tumour mutational burden but with high clonality of immunogenic mutations, expanded T cells, low activation of the WNT pathway and active immune escape mechanisms. Coupling high-dimensional imaging mass cytometry with multiplexed immunofluorescence and computational spatial analysis, we observe that responsive hypermutated CRCs are rich in cytotoxic and proliferating PD1-expressing CD8 cells interacting with high-density clusters of PDL1-expressing antigen presenting macrophages. We propose that anti-PD1 agents release the PD1-PDL1 interaction between CD8 T cells and macrophages thus promoting cytotoxic anti-tumour activity.
]]></description>
<dc:creator>Bortolomeazzi, M.</dc:creator>
<dc:creator>Keddar, M. R.</dc:creator>
<dc:creator>Montorsi, L.</dc:creator>
<dc:creator>Acha-Sagredo, A.</dc:creator>
<dc:creator>Benedetti, L.</dc:creator>
<dc:creator>Temelkovski, D.</dc:creator>
<dc:creator>Choi, S.</dc:creator>
<dc:creator>Petrov, N.</dc:creator>
<dc:creator>Todd, K.</dc:creator>
<dc:creator>Wai, P.</dc:creator>
<dc:creator>Kohl, J.</dc:creator>
<dc:creator>Denner, T.</dc:creator>
<dc:creator>Nye, E.</dc:creator>
<dc:creator>Goldstone, R.</dc:creator>
<dc:creator>Ward, S.</dc:creator>
<dc:creator>Wilson, G. A.</dc:creator>
<dc:creator>Al Bakir, M.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>John, S.</dc:creator>
<dc:creator>Miles, J.</dc:creator>
<dc:creator>Larijani, B.</dc:creator>
<dc:creator>Kunene, V.</dc:creator>
<dc:creator>Fontana, E.</dc:creator>
<dc:creator>Arkenau, H.-T.</dc:creator>
<dc:creator>Parker, P. J.</dc:creator>
<dc:creator>Rodriguez-Justo, M.</dc:creator>
<dc:creator>Shiu, K.-K.</dc:creator>
<dc:creator>Spencer, J.</dc:creator>
<dc:creator>Ciccarelli, F. D.</dc:creator>
<dc:date>2020-12-15</dc:date>
<dc:identifier>doi:10.1101/2020.12.15.422831</dc:identifier>
<dc:title><![CDATA[Immunogenomic profile of colorectal cancer response to immune checkpoint blockade]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.12.16.423054v1?rss=1">
<title>
<![CDATA[
Inhibition of protein N-myristoylation blocks Plasmodium falciparum intraerythrocytic development, egress, and invasion 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.16.423054v1?rss=1"
</link>
<description><![CDATA[
We have combined chemical biology and genetic modification approaches to investigate the importance of protein myristoylation in the human malaria parasite, Plasmodium falciparum. Parasite treatment during schizogony in the last ten to fifteen hours of the erythrocytic cycle with IMP-1002, an inhibitor of N-myristoyl transferase (NMT), led to a significant blockade in parasite egress from the infected erythrocyte. Two rhoptry proteins were mislocalized in the cell, suggesting that rhoptry function is disrupted. We identified sixteen NMT substrates for which myristoylation was significantly reduced by NMT inhibitor treatment, and of these, six proteins were substantially reduced in abundance. In a viability screen, we showed that for four of these proteins replacement of the N-terminal glycine with alanine to prevent myristoylation had a substantial effect on parasite fitness. In detailed studies of one NMT substrate, glideosome associated protein 45 (GAP45), loss of myristoylation had no impact on protein location or glideosome assembly, in contrast to the disruption caused by GAP45 gene deletion, but GAP45 myristoylation was essential for erythrocyte invasion. Therefore, there are at least three mechanisms by which inhibition of NMT can disrupt parasite development and growth: early in parasite development, leading to the inhibition of schizogony and formation of  pseudoschizonts, which has been described previously; at the end of schizogony, with disruption of rhoptry formation, merozoite development and egress from the infected erythrocyte; and at invasion, when impairment of motor complex function prevents invasion of new erythrocytes. These results underline the importance of P. falciparum NMT as a drug target because of the pleiotropic effect of its inhibition.
]]></description>
<dc:creator>Schlott, A. C.</dc:creator>
<dc:creator>Knuepfer, E.</dc:creator>
<dc:creator>Green, J. L.</dc:creator>
<dc:creator>Hobson, P.</dc:creator>
<dc:creator>Borg, A. J.</dc:creator>
<dc:creator>Morales-Sanfrutos, J.</dc:creator>
<dc:creator>Perrin, A. J.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Tate, E. W.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:date>2020-12-16</dc:date>
<dc:identifier>doi:10.1101/2020.12.16.423054</dc:identifier>
<dc:title><![CDATA[Inhibition of protein N-myristoylation blocks Plasmodium falciparum intraerythrocytic development, egress, and invasion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.12.18.423466v1?rss=1">
<title>
<![CDATA[
KLF17 promotes human naïve pluripotency but is not required for its establishment 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.18.423466v1?rss=1"
</link>
<description><![CDATA[
Current knowledge of the transcriptional regulation of human pluripotency is incomplete, with lack of inter-species conservation observed. Single-cell transcriptomics of human embryos previously enabled us to identify transcription factors, including the zinc-finger protein KLF17, that are enriched in the human epiblast and naive hESCs. Here we show that KLF17 is expressed coincident with the known pluripotency factors NANOG and SOX2 across human blastocyst development. We investigate the function of KLF17 in pluripotency using primed and naive hESCs for gain- and loss-of-function analyses. We find that ectopic expression of KLF17 in primed hESCs is sufficient to induce a naive-like transcriptome and that KLF17 can drive transgene-mediated resetting to naive pluripotency. This implies a role for KLF17 in establishing naive pluripotency. However, CRISPR-Cas9-mediated knockout studies reveal that KLF17 is not required for naive pluripotency acquisition in vitro. Transcriptome analysis of naive hESCs identifies subtle effects on metabolism and signalling following KLF17 loss of function, and possible redundancy with the related factor, KLF5. Overall, we show that KLF17 is sufficient, but not necessary, for naive pluripotency under the given in vitro conditions.

Summary statementInvestigating KLF17 in human pluripotency reveals that it is sufficient, but not necessary, to establish naive hESCs. We posit that KLF17 is a peripheral regulator, like KLF2 in the mouse.
]]></description>
<dc:creator>Lea, R. A.</dc:creator>
<dc:creator>McCarthy, A.</dc:creator>
<dc:creator>Boeing, S.</dc:creator>
<dc:creator>Niakan, K. K.</dc:creator>
<dc:date>2020-12-18</dc:date>
<dc:identifier>doi:10.1101/2020.12.18.423466</dc:identifier>
<dc:title><![CDATA[KLF17 promotes human naïve pluripotency but is not required for its establishment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.12.18.423481v1?rss=1">
<title>
<![CDATA[
Molecular fluctuations as a ruler of force-induced protein conformations 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.18.423481v1?rss=1"
</link>
<description><![CDATA[
Molecular fluctuations directly reflect the underlying energy landscape. Variance analysis can probe protein dynamics in several biochemistry-driven approaches, yet measurement of probe-independent fluctuations in proteins exposed to mechanical forces remains only accessible through steered molecular dynamics simulations. Using single molecule magnetic tweezers, here we conduct variance analysis to show that individual unfolding and refolding transitions occurring in dynamic equilibrium in a single protein under force are hallmarked by a change in the protein's end-to-end fluctuations, revealing a change in protein stiffness. By unfolding and refolding three structurally distinct proteins under a wide range of constant forces, we demonstrate that the associated change in protein compliance to reach force-induced thermodynamically-stable states scales with the protein's contour length, in agreement with the sequence-independent FJC model of polymer physics. Our findings will help probe the conformational dynamics of proteins exposed to mechanical force at high resolution, of central importance in mechanosensing and mechanotransduction.
]]></description>
<dc:creator>Stannard, A.</dc:creator>
<dc:creator>Mora, M.</dc:creator>
<dc:creator>Beedle, A. E. M.</dc:creator>
<dc:creator>Castro-Lopez, M.</dc:creator>
<dc:creator>Board, S.</dc:creator>
<dc:creator>Garcia-Manyes, S.</dc:creator>
<dc:date>2020-12-19</dc:date>
<dc:identifier>doi:10.1101/2020.12.18.423481</dc:identifier>
<dc:title><![CDATA[Molecular fluctuations as a ruler of force-induced protein conformations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.12.18.423524v1?rss=1">
<title>
<![CDATA[
Longitudinal omics in Syrian hamsters integrated with human data unravel complexity of moderate immune responses to SARS-CoV-2 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.18.423524v1?rss=1"
</link>
<description><![CDATA[
In COVID-19, the immune response largely determines disease severity and is key to therapeutic strategies. Cellular mechanisms contributing to inflammatory lung injury and tissue repair in SARS-CoV-2 infection, particularly endothelial cell involvement, remain ill-defined. We performed detailed spatiotemporal analyses of cellular and molecular processes in SARS-CoV-2 infected Syrian hamsters. Comparison of hamster single-cell sequencing and proteomics with data sets from COVID-19 patients demonstrated inter-species concordance of cellular and molecular host-pathogen interactions. In depth vascular and pulmonary compartment analyses (i) supported the hypothesis that monocyte-derived macrophages dominate inflammation, (ii) revealed endothelial inflammation status and T-cell attraction, and (iii) showed that CD4+ and CD8+ cytotoxic T-cell responses precede viral elimination. Using the Syrian hamster model of self-limited moderate COVID-19, we defined the specific roles of endothelial and epithelial cells, among other myeloid and non-myeloid lung cell subtypes, for determining the disease course.
]]></description>
<dc:creator>Nouailles, G.</dc:creator>
<dc:creator>Wyler, E.</dc:creator>
<dc:creator>Pennitz, P.</dc:creator>
<dc:creator>Postmus, D.</dc:creator>
<dc:creator>Vladimirova, D.</dc:creator>
<dc:creator>Kazmierski, J.</dc:creator>
<dc:creator>Pott, F.</dc:creator>
<dc:creator>Dietert, K.</dc:creator>
<dc:creator>Mülleder, M.</dc:creator>
<dc:creator>Farztdinov, V.</dc:creator>
<dc:creator>Obermayer, B.</dc:creator>
<dc:creator>Wienhold, S.-M.</dc:creator>
<dc:creator>Andreotti, S.</dc:creator>
<dc:creator>Höfler, T.</dc:creator>
<dc:creator>Sawitzki, B.</dc:creator>
<dc:creator>Drosten, C.</dc:creator>
<dc:creator>Sander, L. E.</dc:creator>
<dc:creator>Suttorp, N.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:creator>Beule, D.</dc:creator>
<dc:creator>Gruber, A. D.</dc:creator>
<dc:creator>Goffinet, C.</dc:creator>
<dc:creator>Landthaler, M.</dc:creator>
<dc:creator>Trimpert, J.</dc:creator>
<dc:creator>Witzenrath, M.</dc:creator>
<dc:date>2020-12-19</dc:date>
<dc:identifier>doi:10.1101/2020.12.18.423524</dc:identifier>
<dc:title><![CDATA[Longitudinal omics in Syrian hamsters integrated with human data unravel complexity of moderate immune responses to SARS-CoV-2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.12.18.423280v1?rss=1">
<title>
<![CDATA[
Targeted cancer therapy induces APOBEC fuelling the evolution of drug resistance 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.18.423280v1?rss=1"
</link>
<description><![CDATA[
Introductory paragraphThe clinical success of targeted cancer therapy is limited by drug resistance that renders cancers lethal in patients1-4. Human tumours can evolve therapy resistance by acquiring de novo genetic alterations and increased heterogeneity via mechanisms that remain incompletely understood1. Here, through parallel analysis of human clinical samples, tumour xenograft and cell line models and murine model systems, we uncover an unanticipated mechanism of therapy-induced adaptation that fuels the evolution of drug resistance. Targeted therapy directed against EGFR and ALK oncoproteins in lung cancer induced adaptations favoring apolipoprotein B mRNA-editing enzyme, catalytic polypeptide (APOBEC)-mediated genome mutagenesis. In human oncogenic EGFR-driven and ALK-driven lung cancers and preclinical models, EGFR or ALK inhibitor treatment induced the expression and DNA mutagenic activity of APOBEC3B via therapy-mediated activation of NF-{kappa}B signaling. Moreover, targeted therapy also mediated downregulation of certain DNA repair enzymes such as UNG2, which normally counteracts APOBEC-catalyzed DNA deamination events. In mutant EGFR-driven lung cancer mouse models, APOBEC3B was detrimental to tumour initiation and yet advantageous to tumour progression during EGFR targeted therapy, consistent with TRACERx data demonstrating subclonal enrichment of APOBEC-mediated mutagenesis. This study reveals how cancers adapt and drive genetic diversity in response to targeted therapy and identifies APOBEC deaminases as future targets for eliciting more durable clinical benefit to targeted cancer therapy.
]]></description>
<dc:creator>Mayekar, M. K.</dc:creator>
<dc:creator>Caswell, D. R.</dc:creator>
<dc:creator>Vokes, N. I.</dc:creator>
<dc:creator>Law, E. K.</dc:creator>
<dc:creator>Wu, W.</dc:creator>
<dc:creator>Hill, W.</dc:creator>
<dc:creator>Gronroos, E.</dc:creator>
<dc:creator>Rowan, A.</dc:creator>
<dc:creator>Al Bakir, M.</dc:creator>
<dc:creator>McCoach, C. E.</dc:creator>
<dc:creator>Blakely, C. M.</dc:creator>
<dc:creator>Temiz, N. A.</dc:creator>
<dc:creator>Nagano, A.</dc:creator>
<dc:creator>Kerr, D. L.</dc:creator>
<dc:creator>Rotow, J. K.</dc:creator>
<dc:creator>Haderk, F.</dc:creator>
<dc:creator>Dietzen, M.</dc:creator>
<dc:creator>Ruiz, C. M.</dc:creator>
<dc:creator>Almeida, B.</dc:creator>
<dc:creator>Cech, L.</dc:creator>
<dc:creator>Gini, B.</dc:creator>
<dc:creator>Przewrocka, J.</dc:creator>
<dc:creator>Moore, C.</dc:creator>
<dc:creator>Murillo, M.</dc:creator>
<dc:creator>Bakker, B.</dc:creator>
<dc:creator>Rule, B.</dc:creator>
<dc:creator>Durfee, C.</dc:creator>
<dc:creator>Nanjo, S.</dc:creator>
<dc:creator>Tan, L.</dc:creator>
<dc:creator>Larson, L. K.</dc:creator>
<dc:creator>Argyris, P. P.</dc:creator>
<dc:creator>Brown, W. L.</dc:creator>
<dc:creator>Yu, J.</dc:creator>
<dc:creator>Gomez, C.</dc:creator>
<dc:creator>Gui, P.</dc:creator>
<dc:creator>Vogel, R. I.</dc:creator>
<dc:creator>Yu, E. A.</dc:creator>
<dc:creator>Thomas, N. J.</dc:creator>
<dc:creator>Venkatesan, S.</dc:creator>
<dc:creator>Hobor, S.</dc:creator>
<dc:creator>Chew, S. K.</dc:creator>
<dc:creator>Kanu, N.</dc:creator>
<dc:creator>McGranahan, N.</dc:creator>
<dc:creator>Van Allen, E. M.</dc:creator>
<dc:creator>Downward, J.</dc:creator>
<dc:creator>Harris, R. S.</dc:creator>
<dc:creator>Bivona, T</dc:creator>
<dc:date>2020-12-18</dc:date>
<dc:identifier>doi:10.1101/2020.12.18.423280</dc:identifier>
<dc:title><![CDATA[Targeted cancer therapy induces APOBEC fuelling the evolution of drug resistance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.08.25.266163v1?rss=1">
<title>
<![CDATA[
A physical theory of larval Drosophila behaviour 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.25.266163v1?rss=1"
</link>
<description><![CDATA[
All animal behaviour must ultimately be governed by physical laws. As a basis for understanding the physics of behaviour in a simple system, we here develop an effective theory for the motion of the larval form of the fruitfly Drosophila melanogaster, and compare it against a quantitative analysis of the real animals behaviour. We first define a set of fields which quantify stretching, bending, and twisting along the larvas antero- posterior axis, and then perform a search in the space of possible theories that could govern the long-wavelength physics of these fields, using a simplified approach inspired by the renormalisation group. Guided by symmetry considerations and stability requirements, we arrive at a unique, analytically tractable free-field theory with a minimum of free parameters. Unexpectedly, we are able to explain a wide-spectrum of features of Drosophila larval behaviour by applying equilibrium statistical mechanics: our theory closely predicts the animals postural modes (eigenmaggots), as well as distributions and trajectories in the postural mode space across several behaviours, including peristaltic crawling, rolling, self-righting and unbiased substrate exploration. We explain the low-dimensionality of postural dynamics via Boltzmann suppression of high frequency modes, and also propose and experimentally test, novel predictions on the relationships between different forms of body deformation and animal behaviour. We show that crawling and rolling are dominated by similar symmetry properties, leading to identical dynamics/statistics in mode space, while rolling and unbiased exploration have a common dominant timescale. Furthermore, we are able to demonstrate that self-righting behaviour occurs continuously throughout substrate exploration, owing to the decoupling of stretching, bending, and twisting at low energies. Together, our results demonstrate that relatively simple effective physics can be used to explain and predict a wide range of animal behaviours.
]]></description>
<dc:creator>Loveless, J.</dc:creator>
<dc:creator>Garner, A.</dc:creator>
<dc:creator>Issa, A. R.</dc:creator>
<dc:creator>Webb, B.</dc:creator>
<dc:creator>Ohyama, T.</dc:creator>
<dc:creator>Alonso, C.</dc:creator>
<dc:date>2020-08-25</dc:date>
<dc:identifier>doi:10.1101/2020.08.25.266163</dc:identifier>
<dc:title><![CDATA[A physical theory of larval Drosophila behaviour]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.12.21.414649v1?rss=1">
<title>
<![CDATA[
Dynamic regulation of hierarchical heterogeneity in Acute Myeloid Leukemia serves as a tumor immunoevasion mechanism 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.21.414649v1?rss=1"
</link>
<description><![CDATA[
Acute Myeloid Leukemia, a hematological malignancy with poor clinical outcome, is composed of hierarchically heterogeneous cells. We examine the contribution of this heterogeneity to disease progression in the context of anti-tumor immune responses and investigate whether these responses regulate the balance between stemness and differentiation in AML. Combining phenotypic analysis with proliferation dynamics and fate-mapping of AML cells in a murine AML model, we demonstrate the presence of a terminally differentiated, chemoresistant population expressing high levels of PDL1. We show that PDL1 upregulation in AML cells, following exposure to IFN{gamma} from activated T cells, is coupled with AML differentiation and the dynamic balance between proliferation, versus differentiation and immunosuppression, facilitates disease progression in the presence of immune responses. This microenvironment-responsive hierarchical heterogeneity in AML may be key in facilitating disease growth at the population level at multiple stages of disease, including following bone marrow transplantation and immunotherapy.
]]></description>
<dc:creator>Pospori, C.</dc:creator>
<dc:creator>Grey, W.</dc:creator>
<dc:creator>Gonzalez-Anton, S.</dc:creator>
<dc:creator>Gibson, S. V.</dc:creator>
<dc:creator>Georgiou, C.</dc:creator>
<dc:creator>Birch, F.</dc:creator>
<dc:creator>Stevens, G.</dc:creator>
<dc:creator>Williams, T.</dc:creator>
<dc:creator>Khorshed, R.</dc:creator>
<dc:creator>Haltalli, M. L. R.</dc:creator>
<dc:creator>Skoufou-Papoutsaki, M.-N.</dc:creator>
<dc:creator>Sloan, K.</dc:creator>
<dc:creator>Encabo, H. H.</dc:creator>
<dc:creator>Hopkins, J.</dc:creator>
<dc:creator>Christodoulidou, C.</dc:creator>
<dc:creator>Stampoulis, D.</dc:creator>
<dc:creator>Hearn-Yates, F.</dc:creator>
<dc:creator>Gribben, J.</dc:creator>
<dc:creator>Stauss, H. J.</dc:creator>
<dc:creator>Chakraverty, R.</dc:creator>
<dc:creator>Bonnet, D.</dc:creator>
<dc:creator>Lo Celso, C. L.</dc:creator>
<dc:date>2020-12-22</dc:date>
<dc:identifier>doi:10.1101/2020.12.21.414649</dc:identifier>
<dc:title><![CDATA[Dynamic regulation of hierarchical heterogeneity in Acute Myeloid Leukemia serves as a tumor immunoevasion mechanism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.12.22.423126v1?rss=1">
<title>
<![CDATA[
APOBEC3B expression generates an immunogenic model of Kras mutant lung cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.22.423126v1?rss=1"
</link>
<description><![CDATA[
Mutations in oncogenes such as KRAS and EGFR cause a high proportion of lung cancers. Drugs targeting these proteins cause tumour regression but ultimately fail to cure these cancers, leading to intense interest in how best to combine them with other treatments, such as immunotherapies. However, preclinical systems for studying the interaction of lung tumours with the host immune system are inadequate, in part due to the low tumour mutational burden in genetically engineered mouse models. Here we set out to develop mouse models of mutant KRAS-driven lung cancer with an elevated tumour mutational burden by expressing the human DNA cytosine deaminase, APOBEC3B, to mimic the mutational signature seen in human lung cancer. This failed to substantially increase clonal tumour mutational burden and autochthonous tumours remained refractory to immunotherapy. However, by establishing clonal cell lines from these tumours we generated an immunogenic syngeneic transplantation model of KRAS mutant lung adenocarcinoma that was sensitive to immunotherapy. Unexpectedly, we found that anti-tumour immune responses were not directed against neoantigens but instead targeted derepressed endogenous retroviral antigens. The ability of KRASG12C inhibitors to cause regression of KRASG12C-expressing versions of these tumours was markedly potentiated by the adaptive immune system, providing a unique opportunity for the study of combinations of targeted and immunotherapies in immune-hot lung cancer.
]]></description>
<dc:creator>de Carne Trecesson, S.</dc:creator>
<dc:creator>Boumelha, J.</dc:creator>
<dc:creator>Law, E.</dc:creator>
<dc:creator>Romero-Clavijo, P.</dc:creator>
<dc:creator>Mugarza, E.</dc:creator>
<dc:creator>Coelho, M.</dc:creator>
<dc:creator>Moore, C.</dc:creator>
<dc:creator>Rana, S.</dc:creator>
<dc:creator>Caswell, D.</dc:creator>
<dc:creator>Murillo, M.</dc:creator>
<dc:creator>Hancock, D.</dc:creator>
<dc:creator>Argyris, P.</dc:creator>
<dc:creator>Brown, W.</dc:creator>
<dc:creator>Durfee, C.</dc:creator>
<dc:creator>Larson, L.</dc:creator>
<dc:creator>Vogel, R.</dc:creator>
<dc:creator>Alejandro Suarez-Bonnet, A.</dc:creator>
<dc:creator>Priestnall, S.</dc:creator>
<dc:creator>East, P.</dc:creator>
<dc:creator>Ross, S.</dc:creator>
<dc:creator>Molina-Arcas, M.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Harris, R.</dc:creator>
<dc:creator>Downward, J.</dc:creator>
<dc:date>2020-12-22</dc:date>
<dc:identifier>doi:10.1101/2020.12.22.423126</dc:identifier>
<dc:title><![CDATA[APOBEC3B expression generates an immunogenic model of Kras mutant lung cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.12.27.423419v1?rss=1">
<title>
<![CDATA[
CKS1-dependent proteostatic regulation has dual roles combating acute myeloid leukemia whilst protecting normal hematopoiesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.27.423419v1?rss=1"
</link>
<description><![CDATA[
Acute myeloid leukemia (AML) is an aggressive hematological disorder comprising a hierarchy of quiescent leukemic stem cells (LSCs) and proliferating blasts with limited self-renewal ability. AML has a dismal prognosis, with extremely low two-year survival rates in the poorest cytogenetic risk patients, primarily due to the failure of intensive chemotherapy protocols unable to deplete LSCs, which reconstitute the disease in vivo, and the significant toxicity towards healthy hematopoietic cells. Whilst much work has been done to identify genetic and epigenetic vulnerabilities in AML LSCs, little is known about protein dynamics and the role of protein degradation in drug resistance and relapse. Here, using a highly specific inhibitor of the SCFSKP2-CKS1 complex, we report a dual role for CKS1-dependent protein degradation in reducing AML blasts in vivo, and importantly depleting LSCs. Whilst many AML LSC targeted therapies show significant toxicity to healthy hematopoiesis, inhibition of CKS1-dependent protein degradation has the opposite effect, protecting normal hematopoietic cells from chemotherapeutic toxicity. Together these findings demonstrate CKS1-dependent proteostasis is key for normal and malignant hematopoiesis.

SignificanceCKS1-dependent protein degradation is a specific vulnerability in AML LSCs. Specific inhibition of SCFSKP2-CKS1 is lethal to CKS1Bhigh AML blasts and all AML LSCs. Normal hematopoiesis is protected from chemotherapeutic toxicity by inhibition of CKS1-dependent protein degradation, substantiating a dual role for CKS1-dependent protein degradation in clinical treatment of AML.
]]></description>
<dc:creator>Grey, W.</dc:creator>
<dc:creator>Rio-Machin, A.</dc:creator>
<dc:creator>Casado-Izquierdo, P.</dc:creator>
<dc:creator>Miettinen, J.</dc:creator>
<dc:creator>Copley, F.</dc:creator>
<dc:creator>Parsons, A.</dc:creator>
<dc:creator>Heckman, C.</dc:creator>
<dc:creator>Cutillas, P.</dc:creator>
<dc:creator>Gribben, J.</dc:creator>
<dc:creator>Fitzgibbon, J.</dc:creator>
<dc:creator>Bonnet, D.</dc:creator>
<dc:date>2020-12-28</dc:date>
<dc:identifier>doi:10.1101/2020.12.27.423419</dc:identifier>
<dc:title><![CDATA[CKS1-dependent proteostatic regulation has dual roles combating acute myeloid leukemia whilst protecting normal hematopoiesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.12.29.424694v1?rss=1">
<title>
<![CDATA[
Wnt/beta-catenin signalling is dispensable for adult neural stem cell homeostasis and activation. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.29.424694v1?rss=1"
</link>
<description><![CDATA[
Adult mouse hippocampal neural stem cells (NSCs) generate new neurons that integrate into existing hippocampal networks and modulate mood and memory. These NSCs are largely quiescent and are stimulated by niche signals to activate and produce neurons. Wnt/{beta}-catenin signalling acts at different steps along the hippocampal neurogenic lineage and has been shown to promote the proliferation of intermediate progenitor cells. However, whether it has a direct role in the regulation of NSCs still remains unclear. Here we used Wnt/{beta}-catenin reporters and transcriptomic data from in vivo and in vitro models to show that both active and quiescent adult NSCs respond to Wnt/{beta}-catenin signalling. Wnt/{beta}-catenin stimulation instructed neuronal differentiation of active NSCs and promoted the activation or differentiation of quiescent NSCs in a dose-dependent manner. However, we found that inhibiting NSCs response to Wnt, by conditionally deleting {beta}-catenin, did not affect their activation or maintenance of their stem cell characteristics. Together, our results indicate that whilst NSCs do respond to Wnt/{beta}-catenin stimulation in a dose-dependent and state-specific manner, Wnt/{beta}-catenin signalling is not cell-autonomously required to maintain NSC homeostasis, which could reconcile some of the contradictions in the literature as to the role of Wnt/{beta}-catenin signalling in adult hippocampal NSCs.
]]></description>
<dc:creator>Austin, S. H. L.</dc:creator>
<dc:creator>Harris, L.</dc:creator>
<dc:creator>Paun, O.</dc:creator>
<dc:creator>Rigo, P.</dc:creator>
<dc:creator>Guillemot, F.</dc:creator>
<dc:creator>Urban Avellaneda, N.</dc:creator>
<dc:date>2020-12-29</dc:date>
<dc:identifier>doi:10.1101/2020.12.29.424694</dc:identifier>
<dc:title><![CDATA[Wnt/beta-catenin signalling is dispensable for adult neural stem cell homeostasis and activation.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.12.29.424684v1?rss=1">
<title>
<![CDATA[
Sox2 levels configure the WNT response of epiblast progenitors responsible for vertebrate body formation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.29.424684v1?rss=1"
</link>
<description><![CDATA[
WNT signalling has multiple roles. It maintains pluripotency of embryonic stem cells, assigns posterior identity in the epiblast and induces mesodermal tissue. We provide evidence that these distinct functions are conducted by the transcription factor SOX2, which adopts different modes of chromatin interaction and regulatory element selection depending on its level of expression. At high levels, SOX2 acts as a pioneer factor, displacing nucleosomes from regulatory elements with high affinity SOX2 binding sites and recruiting the WNT effector, TCF/{beta}-catenin, to maintain pluripotent gene expression. Reducing SOX2 levels destabilises pluripotency and reconfigures SOX2/TCF/{beta}-catenin occupancy to caudal epiblast expressed genes. These contain low-affinity SOX2 sites and are co-occupied by T/Bra and CDX. The loss of SOX2 allows WNT induced mesodermal differentiation. These findings define a role for Sox2 levels in dictating the chromatin occupancy of TCF/{beta}-catenin and reveal how context specific responses to a signal are configured by the level of a transcription factor.
]]></description>
<dc:creator>Blassberg, R.</dc:creator>
<dc:creator>Patel, H.</dc:creator>
<dc:creator>Watson, T.</dc:creator>
<dc:creator>Gouti, M.</dc:creator>
<dc:creator>Metzis, V.</dc:creator>
<dc:creator>Delas, J.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:date>2020-12-30</dc:date>
<dc:identifier>doi:10.1101/2020.12.29.424684</dc:identifier>
<dc:title><![CDATA[Sox2 levels configure the WNT response of epiblast progenitors responsible for vertebrate body formation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.12.31.424931v1?rss=1">
<title>
<![CDATA[
Structural insights into Cullin4-RING ubiquitin ligase remodelling by Vpr from simian immunodeficiency viruses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.31.424931v1?rss=1"
</link>
<description><![CDATA[
Viruses have evolved means to manipulate the hosts ubiquitin-proteasome system, in order to down-regulate antiviral host factors. The Vpx/Vpr family of lentiviral accessory proteins usurp the substrate receptor DCAF1 of host Cullin4-RING ligases (CRL4), a family of modular ubiquitin ligases involved in DNA replication, DNA repair and cell cycle regulation. CRL4DCAF1 specificity modulation by Vpx and Vpr from certain simian immunodeficiency viruses (SIV) leads to recruitment, poly-ubiquitylation and subsequent proteasomal degradation of the host restriction factor SAMHD1, resulting in enhanced virus replication in differentiated cells. To unravel the mechanism of SIV Vpr-induced SAMHD1 ubiquitylation, we conducted integrative biochemical and structural analyses of the Vpr protein from SIVs infecting Cercopithecus cephus (SIVmus). X-ray crystallography reveals commonalities between SIVmus Vpr and other members of the Vpx/Vpr family with regard to DCAF1 interaction, while cryo-electron microscopy and cross-linking mass spectrometry highlight a divergent molecular mechanism of SAMHD1 recruitment. In addition, these studies demonstrate how SIVmus Vpr exploits the dynamic architecture of the multi-subunit CRL4DCAF1 assembly to optimise SAMHD1 ubiquitylation. Together, the present work provides detailed molecular insight into variability and species-specificity of the evolutionary arms race between host SAMHD1 restriction and lentiviral counteraction through Vpx/Vpr proteins.

Author summaryDue to the limited size of virus genomes, virus replication critically relies on host cell components. In addition to the host cells energy metabolism and its DNA replication and protein synthesis apparatus, the protein degradation machinery is an attractive target for viral re-appropriation. Certain viral factors divert the specificity of host ubiquitin ligases to antiviral host factors, in order to mark them for destruction by the proteasome, to lift intracellular barriers to virus replication. Here, we present molecular details of how the simian immunodeficiency virus accessory protein Vpr interacts with a substrate receptor of host Cullin4-RING ubiquitin ligases, and how this interaction redirects the specificity of Cullin4-RING to the antiviral host factor SAMHD1. The studies uncover the mechanism of Vpr-induced SAMHD1 recruitment and subsequent ubiquitylation. Moreover, by comparison to related accessory proteins from other immunodeficiency virus species, we illustrate the surprising variability in the molecular strategies of SAMHD1 counteraction, which these viruses adopted during evolutionary adaptation to their hosts. Lastly, our work also provides deeper insight into the inner workings of the hosts Cullin4-RING ubiquitylation machinery.
]]></description>
<dc:creator>Banchenko, S.</dc:creator>
<dc:creator>Krupp, F.</dc:creator>
<dc:creator>Gotthold, C.</dc:creator>
<dc:creator>Buerger, J.</dc:creator>
<dc:creator>Graziadei, A.</dc:creator>
<dc:creator>O' Reilly, F.</dc:creator>
<dc:creator>Sinn, L.</dc:creator>
<dc:creator>Ruda, O.</dc:creator>
<dc:creator>Rappsilber, J.</dc:creator>
<dc:creator>Spahn, C. M. T.</dc:creator>
<dc:creator>Mielke, T.</dc:creator>
<dc:creator>Taylor, I. A.</dc:creator>
<dc:creator>Schwefel, D.</dc:creator>
<dc:date>2021-01-01</dc:date>
<dc:identifier>doi:10.1101/2020.12.31.424931</dc:identifier>
<dc:title><![CDATA[Structural insights into Cullin4-RING ubiquitin ligase remodelling by Vpr from simian immunodeficiency viruses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.01.05.425371v1?rss=1">
<title>
<![CDATA[
Single-molecule super-resolution imaging of T-cell plasma membrane CD4 redistribution upon HIV-1 binding 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.01.05.425371v1?rss=1"
</link>
<description><![CDATA[
The first step of cellular entry for the human immunodeficiency virus type-1 (HIV-1) occurs through the binding of its envelope protein (Env) with the plasma membrane receptor CD4 and co-receptor CCR5 or CXCR4 on susceptible cells, primarily CD4+ T cells and macrophages. Although there is considerable knowledge of the molecular interactions between Env and host cell receptors that lead to successful fusion, the precise way in which HIV-1 receptors redistribute to sites of virus binding at the nanoscale remains unknown. Here, we quantitatively examine changes in the nanoscale organisation of CD4 on the surface of CD4+ T cells following HIV-1 binding. Using single-molecule super-resolution imaging, we show that CD4 molecules are distributed mostly as either individual molecules or small clusters of up to 4 molecules. Following virus binding, we observe a local 3-to-10-fold increase in cluster diameter and molecule number for virus-associated CD4 clusters. Moreover, a similar but smaller magnitude reorganisation of CD4 was also observed with recombinant gp120. For the first time, our results quantify the nanoscale CD4 reorganisation triggered by HIV-1 on host cells. Our quantitative approach provides a robust methodology for characterising the nanoscale organisation of plasma membrane receptors in general with the potential to link spatial organisation to function.
]]></description>
<dc:creator>Yuan, Y.</dc:creator>
<dc:creator>Jacobs, C.</dc:creator>
<dc:creator>Llorente Garcia, I.</dc:creator>
<dc:creator>Pereira, P. M.</dc:creator>
<dc:creator>Lawrence, S.</dc:creator>
<dc:creator>Laine, R. F.</dc:creator>
<dc:creator>Marsh, M.</dc:creator>
<dc:creator>Henriques, R.</dc:creator>
<dc:date>2021-01-05</dc:date>
<dc:identifier>doi:10.1101/2021.01.05.425371</dc:identifier>
<dc:title><![CDATA[Single-molecule super-resolution imaging of T-cell plasma membrane CD4 redistribution upon HIV-1 binding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.01.04.425361v1?rss=1">
<title>
<![CDATA[
Spontaneous selection of Cryptosporidium drug resistance in a calf model of infection 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.01.04.425361v1?rss=1"
</link>
<description><![CDATA[
The intestinal protozoan Cryptosporidium is a leading cause of diarrheal disease and mortality in young children. There is currently no fully effective treatment for cryptosporidiosis, which has stimulated interest in anticryptosporidial development over the last [~]10 years with numerous lead compounds identified including several tRNA synthetase inhibitors. In this study, we report the results of a dairy calf efficacy trial of the methionyl-tRNA (CpMetRS) synthetase inhibitor 2093 and the spontaneous emergence of drug resistance. Dairy calves experimentally infected with Cryptosporidium parvum initially improved with 2093 treatment, but parasite shedding resumed in two of three calves on treatment day five. Parasites shed by each recrudescent calf had different amino acid altering CpMetRS mutations, coding either an aspartate 243 to glutamate (D243E) or a threonine 246 to isoleucine (T246I) mutation. Transgenic parasites engineered to have either the D243E or T246I CpMetRS mutation using CRISPR/Cas9 grew normally but were highly 2093 resistant; the D243E and T246I mutant expressing parasites respectively had 2093 EC50S of 613- or 128-fold that of transgenic parasites with wild-type CpMetRS. In studies using recombinant enzymes, the D243E and T246I mutations shifted the 2093 IC50 by >170-fold. Structural modeling of CpMetRS based on an inhibitor-bound Trypanosoma brucei MetRS crystal structure suggested that the resistance mutations reposition nearby hydrophobic residues, interfering with compound binding while minimally impacting substrate binding. This is the first report of naturally emerging Cryptosporidium drug resistance, highlighting the need to address the potential for anticryptosporidial resistance and establish strategies to limit its occurrence.

ImportanceCryptosporidium is a leading protozoan cause of diarrhea in young children with no reliable treatment. We report results of a dairy calf drug efficacy trial and the spontaneous emergence of drug resistance. Cryptosporidium parvum infected calves initially improved with drug treatment, but infection relapsed in two animals. Parasites shed by each recrudescent calf had mutations in the gene encoding the drug target that altered its amino acid sequence. Recapitulation of the drug target mutations by CRISPR/Cas9 genome editing resulted in highly drug-resistant parasites, and recombinant mutant enzymes were resistant to inhibition. This is the first report of naturally emerging Cryptosporidium drug resistance. There is a currently a great opportunity to impact public health with new drugs to treat cryptosporidiosis, and this report highlights the need to address the potential for anticryptosporidial resistance and establish strategies to limit its occurrence in order to realize their full potential.

One-sentence summaryDrug-target point mutations mediating anticryptosporidial resistance spontaneously arose in the dairy calf C. parvum infection model.
]]></description>
<dc:creator>Hasan, M. M.</dc:creator>
<dc:creator>Stebbins, E. E.</dc:creator>
<dc:creator>Choy, R. K. M.</dc:creator>
<dc:creator>Gillespie, J. R.</dc:creator>
<dc:creator>de Hostos, E.</dc:creator>
<dc:creator>Miller, P. J.</dc:creator>
<dc:creator>Mushtaq, A.</dc:creator>
<dc:creator>Ranade, R. M.</dc:creator>
<dc:creator>Teixeira, J. E.</dc:creator>
<dc:creator>Verlinde, C. L. M. J.</dc:creator>
<dc:creator>Sateriale, A.</dc:creator>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Osbourn, D. M.</dc:creator>
<dc:creator>Griggs, D. W.</dc:creator>
<dc:creator>Fan, E.</dc:creator>
<dc:creator>Buckner, F. S.</dc:creator>
<dc:creator>Huston, C. D.</dc:creator>
<dc:date>2021-01-05</dc:date>
<dc:identifier>doi:10.1101/2021.01.04.425361</dc:identifier>
<dc:title><![CDATA[Spontaneous selection of Cryptosporidium drug resistance in a calf model of infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.12.19.423464v1?rss=1">
<title>
<![CDATA[
Free recall scaling laws and short-term memory effects in a latching attractor network 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.19.423464v1?rss=1"
</link>
<description><![CDATA[
Despite the complexity of human memory, paradigms like free recall have revealed robust qualitative and quantitative characteristics, such as power laws governing recall capacity. Although abstract random matrix models could explain such laws, the possibility of their implementation in large networks of interacting neurons has so far remained unexplored. We study an attractor network model of long-term memory endowed with firing rate adaptation and global inhibition. Under appropriate conditions, the transitioning behaviour of the network from memory to memory is constrained by limit cycles that prevent the network from recalling all memories, with scaling similar to what has been found in experiments. When the model is supplemented with a heteroassociative learning rule, complementing the standard autoassociative learning rule, as well as short-term synaptic facilitation, our model reproduces other key findings in the free recall literature, namely serial position effects, contiguity and forward asymmetry effects, as well as the semantic effects found to guide memory recall. The model is consistent with a broad series of manipulations aimed at gaining a better understanding of the variables that affect recall, such as the role of rehearsal, presentation rates and (continuous/end-of-list) distractor conditions. We predict that recall capacity may be increased with the addition of small amounts of noise, for example in the form of weak random stimuli during recall. Moreover, we predict that although the statistics of the encoded memories has a strong effect on the recall capacity, the power laws governing recall capacity may still be expected to hold.
]]></description>
<dc:creator>Boboeva, V.</dc:creator>
<dc:creator>Pezzotta, A.</dc:creator>
<dc:creator>Clopath, C.</dc:creator>
<dc:date>2020-12-21</dc:date>
<dc:identifier>doi:10.1101/2020.12.19.423464</dc:identifier>
<dc:title><![CDATA[Free recall scaling laws and short-term memory effects in a latching attractor network]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.01.06.425462v1?rss=1">
<title>
<![CDATA[
Neural stem cells alter nucleocytoplasmic partitioning and accumulate nuclear polyadenylated transcripts during quiescence 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.01.06.425462v1?rss=1"
</link>
<description><![CDATA[
Quiescence is a cellular state characterised by reversible cell-cycle arrest and diminished biosynthetic activity that protects against environmental insults, replicative exhaustion and proliferation-induced mutations1. Entry into and exit from this state controls development, maintenance and repair of tissues plus, in the adult central nervous system, generation of new neurons and thus cognition and mood2-4. Cancer stem cells too can undergo quiescence, which confers them resistance to current therapies5, 6. Despite clinical relevance, quiescence is poorly understood and is defined functionally given lack of molecular markers. Decrease of the most resource-intensive cellular process of protein synthesis is a feature of quiescence, controlled across species and cell types by inhibition of the Target of Rapamycin (TOR) pathway1, 7. Here, we combine Drosophila genetics and a mammalian model to show that altered nucleocytoplasmic partitioning and nuclear accumulation of polyadenylated RNAs are novel evolutionarily conserved hallmarks of quiescence regulation. Furthermore, nuclear accumulation of messenger RNA (mRNA) in quiescent NSCs (qNSCs) largely predicts protein downregulation, accounting for uncoupling between transcriptome and proteome in quiescence. These mechanisms provide a previously unappreciated regulatory layer to reducing protein synthesis in quiescent cells, whilst priming them for reactivation in response to appropriate cues.
]]></description>
<dc:creator>Rossi, A.</dc:creator>
<dc:creator>Coum, A.</dc:creator>
<dc:creator>Madelenat, M.</dc:creator>
<dc:creator>Harris, L.</dc:creator>
<dc:creator>Miedzik, A.</dc:creator>
<dc:creator>Strohbuecker, S.</dc:creator>
<dc:creator>Chai, A.</dc:creator>
<dc:creator>Fiaz, H.</dc:creator>
<dc:creator>Chaouni, R.</dc:creator>
<dc:creator>Faull, P.</dc:creator>
<dc:creator>Grey, W.</dc:creator>
<dc:creator>Bonnet, D.</dc:creator>
<dc:creator>Makeyev, E. V.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Kelly, G.</dc:creator>
<dc:creator>Guillemot, F.</dc:creator>
<dc:creator>Sousa-Nunes, R.</dc:creator>
<dc:date>2021-01-06</dc:date>
<dc:identifier>doi:10.1101/2021.01.06.425462</dc:identifier>
<dc:title><![CDATA[Neural stem cells alter nucleocytoplasmic partitioning and accumulate nuclear polyadenylated transcripts during quiescence]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.01.08.425895v1?rss=1">
<title>
<![CDATA[
Genome-scale sequencing and analysis of human, wolf and bison DNA from 25,000 year-old sediment 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.01.08.425895v1?rss=1"
</link>
<description><![CDATA[
Archaeological sediments have been shown to preserve ancient DNA, but so far have not yielded genome-scale information of the magnitude of skeletal remains. We retrieved and analysed human and mammalian low-coverage nuclear and high-coverage mitochondrial genomes from Upper Palaeolithic sediments from Satsurblia cave, western Georgia, dated to 25,000 years ago. First, a human female genome with substantial basal Eurasian ancestry, which was an ancestry component of the majority of post-Ice Age people in the Near East, North Africa, and parts of Europe. Second, a wolf genome that is basal to extant Eurasian wolves and dogs and represents a previously unknown, likely extinct, Caucasian lineage that diverged from the ancestors of modern wolves and dogs before these diversified. Third, a bison genome that is basal to present-day populations, suggesting that population structure has been substantially reshaped since the Last Glacial Maximum. Our results provide new insights into the late Pleistocene genetic histories of these three species, and demonstrate that sediment DNA can be used not only for species identification, but also be a source of genome-wide ancestry information and genetic history.

HighlightsO_LIWe demonstrate for the first time that genome sequencing from sediments is comparable to that of skeletal remains
C_LIO_LIA single Pleistocene sediment sample from the Caucasus yielded three low-coverage mammalian ancient genomes
C_LIO_LIWe show that sediment ancient DNA can reveal important aspects of the human and faunal past
C_LIO_LIEvidence of an uncharacterized human lineage from the Caucasus before the Last Glacial Maximum
C_LIO_LI[~]0.01-fold coverage wolf and bison genomes are both basal to present-day diversity, suggesting reshaping of population structure in both species
C_LI
]]></description>
<dc:creator>Gelabert, P.</dc:creator>
<dc:creator>Sawyer, S.</dc:creator>
<dc:creator>Bergstrom, A.</dc:creator>
<dc:creator>Collin, T. C.</dc:creator>
<dc:creator>Meshvelian, T.</dc:creator>
<dc:creator>Belfer-Cohen, A.</dc:creator>
<dc:creator>Lordkipanidze, D.</dc:creator>
<dc:creator>Jakeli, N.</dc:creator>
<dc:creator>Matskevich, Z.</dc:creator>
<dc:creator>Bar-Oz, G.</dc:creator>
<dc:creator>Fernandes, D. M.</dc:creator>
<dc:creator>Cheronet, O.</dc:creator>
<dc:creator>Ozdogan, K. T.</dc:creator>
<dc:creator>Oberreiter, V.</dc:creator>
<dc:creator>Feeney, R. N. M.</dc:creator>
<dc:creator>Skoglund, P.</dc:creator>
<dc:creator>Pinhasi, R.</dc:creator>
<dc:date>2021-01-08</dc:date>
<dc:identifier>doi:10.1101/2021.01.08.425895</dc:identifier>
<dc:title><![CDATA[Genome-scale sequencing and analysis of human, wolf and bison DNA from 25,000 year-old sediment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.01.13.426503v1?rss=1">
<title>
<![CDATA[
Functional and multiscale 3D structural investigation of brain tissue through correlative in vivo physiology, synchrotron micro-tomography and volume electron microscopy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.01.13.426503v1?rss=1"
</link>
<description><![CDATA[
Attributing in vivo neurophysiology to the brains ultrastructure requires a large field of view containing contextual anatomy. Electron microscopy (EM) is the gold standard technique to identify ultrastructure, yet acquiring volumes containing full mammalian neural circuits is challenging and time consuming using EM. Here, we show that synchrotron X-ray computed tomography (SXRT) provides rapid imaging of EM-prepared tissue volumes of several cubic millimetres. Resolution was sufficient for distinguishing cell bodies as well as for tracing apical dendrites in olfactory bulb and hippocampus, for up to 350 m. Correlating EM with SXRT allowed us to associate dendritic spines on pyramidal cell apical dendrites in the stratum radiatum to their corresponding soma locations. Superficial pyramidal neurons had larger spine apparatus density compared to deeper ones, implying differential synaptic plasticity for superficial and deeper cells. Finally, we show that X-ray tomography and volume EM can be reliably correlated to prior in vivo imaging. Thus, combining functional measurements with multiscale X-ray microscopy and volume EM establishes a correlative workflow that enables functional and structural investigation of subcellular features in the context of cellular morphologies, tissues and ultimately whole organs.
]]></description>
<dc:creator>Bosch, C.</dc:creator>
<dc:creator>Ackels, T.</dc:creator>
<dc:creator>Pacureanu, A.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Peddie, C. J.</dc:creator>
<dc:creator>Berning, M.</dc:creator>
<dc:creator>Rzepka, N.</dc:creator>
<dc:creator>Zdora, M.-C.</dc:creator>
<dc:creator>Whiteley, I.</dc:creator>
<dc:creator>Storm, M.</dc:creator>
<dc:creator>Bonnin, A.</dc:creator>
<dc:creator>Rau, C.</dc:creator>
<dc:creator>Margrie, T.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:creator>Schaefer, A. T.</dc:creator>
<dc:date>2021-01-14</dc:date>
<dc:identifier>doi:10.1101/2021.01.13.426503</dc:identifier>
<dc:title><![CDATA[Functional and multiscale 3D structural investigation of brain tissue through correlative in vivo physiology, synchrotron micro-tomography and volume electron microscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.01.15.426883v1?rss=1">
<title>
<![CDATA[
Protein Phosphatase 1 regulates atypical chromosome segregation and cell polarity during mitotic and meiotic division in Plasmodium sexual stages 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.01.15.426883v1?rss=1"
</link>
<description><![CDATA[
PP1 is a conserved eukaryotic serine/threonine phosphatase that regulates many aspects of mitosis and meiosis, often working in concert with other phosphatases, such as CDC14 and CDC25. The proliferative stages of the malaria parasite life cycle include sexual development within the mosquito vector, with male gamete formation characterized by an atypical rapid mitosis, consisting of three rounds of DNA synthesis, successive spindle formation with clustered kinetochores, and a meiotic stage during zygote to ookinete development following fertilization. It is unclear how PP1 is involved in these unusual processes. Using real-time live-cell and ultrastructural imaging, conditional gene knockdown, RNA-seq and proteomic approaches, we show that Plasmodium PP1 is implicated in both mitotic exit and, potentially, establishing cell polarity during zygote development in the mosquito midgut, suggesting that small molecule inhibitors of PP1 should be explored for blocking parasite transmission.
]]></description>
<dc:creator>Zeeshan, M.</dc:creator>
<dc:creator>Pandey, R.</dc:creator>
<dc:creator>SUBUDHI, A. K.</dc:creator>
<dc:creator>Ferguson, D. J. P.</dc:creator>
<dc:creator>Kaur, G.</dc:creator>
<dc:creator>Rashpa, R.</dc:creator>
<dc:creator>Nugmanova, R.</dc:creator>
<dc:creator>Brady, D.</dc:creator>
<dc:creator>Bottrill, A.</dc:creator>
<dc:creator>vaughan, s.</dc:creator>
<dc:creator>Brochet, M.</dc:creator>
<dc:creator>Bollen, M.</dc:creator>
<dc:creator>Pain, A.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:creator>Guttery, D. S.</dc:creator>
<dc:creator>Tewari, R.</dc:creator>
<dc:date>2021-01-17</dc:date>
<dc:identifier>doi:10.1101/2021.01.15.426883</dc:identifier>
<dc:title><![CDATA[Protein Phosphatase 1 regulates atypical chromosome segregation and cell polarity during mitotic and meiotic division in Plasmodium sexual stages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.01.15.426849v1?rss=1">
<title>
<![CDATA[
The impact of Spike mutations on SARS-CoV-2 neutralization 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.01.15.426849v1?rss=1"
</link>
<description><![CDATA[
Multiple SARS-CoV-2 vaccines have shown protective efficacy, which is most likely mediated by neutralizing antibodies recognizing the viral entry protein, Spike. Antibodies from SARS-CoV-2 infection neutralize the virus by focused targeting of Spike and there is limited serum cross-neutralization of the closely-related SARS-CoV. As new SARS-CoV-2 variants are rapidly emerging, exemplified by the B.1.1.7, 501Y.V2 and P.1 lineages, it is critical to understand if antibody responses induced by infection with the original SARS-CoV-2 virus or the current vaccines will remain effective against virus variants. In this study we evaluate neutralization of a series of mutated Spike pseudotypes including a B.1.1.7 Spike pseudotype. The analyses of a panel of Spike-specific monoclonal antibodies revealed that the neutralizing activity of some antibodies was dramatically reduced by Spike mutations. In contrast, polyclonal antibodies in the serum of patients infected in early 2020 remained active against most mutated Spike pseudotypes. The majority of serum samples were equally able to neutralize the B.1.1.7 Spike pseudotype, however potency was reduced in a small number of samples (3 of 36) by 5-10-fold. This work highlights that changes in the SARS-CoV-2 Spike can alter neutralization sensitivity and underlines the need for effective real-time monitoring of emerging mutations and their impact on vaccine efficacy.
]]></description>
<dc:creator>Rees-Spear, C.</dc:creator>
<dc:creator>Muir, L.</dc:creator>
<dc:creator>Griffith, S. A.</dc:creator>
<dc:creator>Heaney, J.</dc:creator>
<dc:creator>Aldon, Y.</dc:creator>
<dc:creator>Snitselaar, J.</dc:creator>
<dc:creator>Thomas, P.</dc:creator>
<dc:creator>Graham, C.</dc:creator>
<dc:creator>Seow, J.</dc:creator>
<dc:creator>Lee, N.</dc:creator>
<dc:creator>Rosa, A.</dc:creator>
<dc:creator>Roustan, C.</dc:creator>
<dc:creator>Houlihan, C. F.</dc:creator>
<dc:creator>Sanders, R. W.</dc:creator>
<dc:creator>Gupta, R. K.</dc:creator>
<dc:creator>Cherepanov, P.</dc:creator>
<dc:creator>Stauss, H.</dc:creator>
<dc:creator>Nastouli, E.</dc:creator>
<dc:creator>Doores, K. J.</dc:creator>
<dc:creator>van Gils, M. J.</dc:creator>
<dc:creator>McCoy, L. E.</dc:creator>
<dc:date>2021-01-19</dc:date>
<dc:identifier>doi:10.1101/2021.01.15.426849</dc:identifier>
<dc:title><![CDATA[The impact of Spike mutations on SARS-CoV-2 neutralization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.01.19.427232v1?rss=1">
<title>
<![CDATA[
Luminal nutrients activate distinct patterns in submucosal and myenteric neurons in the mouse small intestine 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.01.19.427232v1?rss=1"
</link>
<description><![CDATA[
Nutrient signals sensed by enteroendocrine cells are conveyed to the enteric nervous system (ENS) to initiate intestinal reflexes. We addressed whether there are specific enteric pathways dedicated to detecting different luminal nutrients. Calcium imaging was performed on intact jejunal preparations from Wnt1-cre;R26R-GCaMP3 and Villin-cre;R26R-GCaMP3 mice which express a fluorescent calcium indicator in their ENS or intestinal epithelium, respectively. Glucose, acetate, and L-phenylalanine were perfused onto the mucosa whilst imaging underlying enteric neurons. Nutrient transport or diffusion across the mucosa was mimicked by applying nutrients onto sensory nerve endings in a villus, or onto myenteric ganglia. The nutrients perfused onto the mucosa each elicited Ca2+ transients in submucosal neurons and in distinct patterns of myenteric neurons. Notably, the neurochemical subtypes of myenteric neurons that responded differed between the nutrients, while submucosal responders were predominantly cholinergic. Nutrients applied into villi or onto ganglia did not elicit specific neuronal responses but did stimulate Ca2+ signaling in the mucosal epithelium. These data suggest that nutrients are first detected at the level of the epithelium and that the ENS is capable of discriminating between different compositions of luminal content. Furthermore, our data show that responses to mucosal stimulation are primarily in the myenteric plexus and submucosal neurons respond secondarily.
]]></description>
<dc:creator>Fung, C.</dc:creator>
<dc:creator>Hao, M. M.</dc:creator>
<dc:creator>Obata, Y.</dc:creator>
<dc:creator>Tack, J.</dc:creator>
<dc:creator>Pachnis, V.</dc:creator>
<dc:creator>Boesmans, W.</dc:creator>
<dc:creator>Vanden Berghe, P.</dc:creator>
<dc:date>2021-01-20</dc:date>
<dc:identifier>doi:10.1101/2021.01.19.427232</dc:identifier>
<dc:title><![CDATA[Luminal nutrients activate distinct patterns in submucosal and myenteric neurons in the mouse small intestine]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.01.27.428555v1?rss=1">
<title>
<![CDATA[
Diminished miRNA activity is associated with aberrant cytoplasmic intron retention in ALS pathogenesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.01.27.428555v1?rss=1"
</link>
<description><![CDATA[
Intron retention (IR) is now recognized as a dominant splicing event during motor neuron (MN) development, however the role and regulation of intron-retaining transcripts (IRTs) localized to the cytoplasm remain particularly understudied. By resolving the spatiotemporal dynamics of IR underlying distinct stages of MN lineage restriction, we identify a cytoplasmic group of IRTs that is not associated with reduced expression of their own genes but instead with an upregulation of predicted target genes of specific miRNAs, the motifs of which are enriched within the intronic sequences of this group. Next, we show that ALS-causing VCP mutations lead to a selective increase in IR of this particular class of introns. This in turn temporally coincides with an increase in the expression level of predicted target genes of these miRNAs, providing a potential mechanistic insight into ALS pathogenesis. Altogether, we propose a novel role for the cytoplasmic intronic sequences in regulating miRNA activity through miRNA sequestration, which potentially contributes to ALS pathogenesis.
]]></description>
<dc:creator>Petric-Howe, M.</dc:creator>
<dc:creator>Crerar, H.</dc:creator>
<dc:creator>Neeves, J.</dc:creator>
<dc:creator>Tyzack, G. E.</dc:creator>
<dc:creator>Patani, R.</dc:creator>
<dc:creator>Luisier, R.</dc:creator>
<dc:date>2021-01-28</dc:date>
<dc:identifier>doi:10.1101/2021.01.27.428555</dc:identifier>
<dc:title><![CDATA[Diminished miRNA activity is associated with aberrant cytoplasmic intron retention in ALS pathogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.01.29.428632v1?rss=1">
<title>
<![CDATA[
The regulatory landscape of cells in the developing mouse cerebellum 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.01.29.428632v1?rss=1"
</link>
<description><![CDATA[
Organ development is orchestrated by cell- and time-specific gene regulatory networks. Here we investigated the regulatory basis of mouse cerebellum development from early neurogenesis to adulthood. By acquiring snATAC-seq profiles for ~90,000 cells spanning eleven stages, we mapped all major cerebellar cell types and identified candidate cis-regulatory elements (CREs). We detected extensive spatiotemporal heterogeneity among progenitor cells and characterized the regulatory programs underlying the differentiation of cerebellar neurons. Although CRE activity is predominantly cell type- and time-specific, periods of greater regulatory change are shared across cell types. There is a universal decrease in CRE conservation and pleiotropy during development and differentiation, but the degree of evolutionary constraint differs between cerebellar cell types. Our work delineates the developmental and evolutionary dynamics of gene regulation in cerebellar cells and provides general insights into mammalian organ development.
]]></description>
<dc:creator>Sarropoulos, I.</dc:creator>
<dc:creator>Sepp, M.</dc:creator>
<dc:creator>Frömel, R.</dc:creator>
<dc:creator>Leiss, K.</dc:creator>
<dc:creator>Trost, N.</dc:creator>
<dc:creator>Leushkin, E.</dc:creator>
<dc:creator>Okonechnikov, K.</dc:creator>
<dc:creator>Joshi, P.</dc:creator>
<dc:creator>Kutscher, L. M.</dc:creator>
<dc:creator>Cardoso-Moreira, M.</dc:creator>
<dc:creator>Pfister, S. M.</dc:creator>
<dc:creator>Kaessmann, H.</dc:creator>
<dc:date>2021-01-29</dc:date>
<dc:identifier>doi:10.1101/2021.01.29.428632</dc:identifier>
<dc:title><![CDATA[The regulatory landscape of cells in the developing mouse cerebellum]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.01.31.429033v1?rss=1">
<title>
<![CDATA[
Dendritic domain-specific sampling of long-range axons shapes feedforward and feedback connectivity of L5 neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.01.31.429033v1?rss=1"
</link>
<description><![CDATA[
Feedforward and feedback pathways interact in specific dendritic domains to enable cognitive functions such as predictive processing and learning. Based on axonal projections, hierarchically lower areas are thought to form synapses primarily on dendrites in middle cortical layers, while higher-order areas are posited to target dendrites in layer 1 and in deep layers. However, the extent to which functional synapses form in regions of axo-dendritic overlap has not been extensively studied. Here, we use viral tracing in the secondary visual cortex of mice to map brain-wide inputs to thick-tufted layer 5 pyramidal neurons. Furthermore, we provide a comprehensive map of input locations through subcellular optogenetic circuit mapping. We show that input pathways target distinct dendritic domains with far greater specificity than appears from their axonal branching, often deviating substantially from the canonical patterns. Common assumptions regarding the dendrite-level interaction of feedforward and feedback inputs may thus need revisiting.
]]></description>
<dc:creator>Galloni, A. R.</dc:creator>
<dc:creator>Ye, Z.</dc:creator>
<dc:creator>Rancz, E.</dc:creator>
<dc:date>2021-02-01</dc:date>
<dc:identifier>doi:10.1101/2021.01.31.429033</dc:identifier>
<dc:title><![CDATA[Dendritic domain-specific sampling of long-range axons shapes feedforward and feedback connectivity of L5 neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.02.02.429358v1?rss=1">
<title>
<![CDATA[
Characterisation of tumour immune microenvironment remodelling following oncogene inhibition in preclinical studies using an optimised imaging mass cytometry workflow 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.02.02.429358v1?rss=1"
</link>
<description><![CDATA[
Mouse models are critical in pre-clinical studies of cancer therapy, allowing dissection of mechanisms through chemical and genetic manipulations that are not feasible in the clinical setting. In studies of the tumour microenvironment (TME), multiplexed imaging methods can provide a rich source of information. However, the application of such technologies in mouse tissues is still in its infancy. Here we present a workflow for studying the TME using imaging mass cytometry with a panel of 27 antibodies on frozen mouse tissues. We optimise and validate image segmentation strategies and automate the process in a Nextflow-based pipeline (imcyto) that is scalable and portable, allowing for parallelised segmentation of large multi-image datasets. With these methods we interrogate the remodelling of the TME induced by a KRAS G12C inhibitor in an immune competent mouse orthotopic lung cancer model, highlighting the infiltration and activation of antigen presenting cells and effector cells.
]]></description>
<dc:creator>van Maldegem, F.</dc:creator>
<dc:creator>Valand, K.</dc:creator>
<dc:creator>Cole, M.</dc:creator>
<dc:creator>Patel, H.</dc:creator>
<dc:creator>Angelova, M.</dc:creator>
<dc:creator>Rana, S.</dc:creator>
<dc:creator>Colliver, E.</dc:creator>
<dc:creator>Enfield, K.</dc:creator>
<dc:creator>Bah, N.</dc:creator>
<dc:creator>Tsang, V.</dc:creator>
<dc:creator>Mugarza, E.</dc:creator>
<dc:creator>Moore, C.</dc:creator>
<dc:creator>Hobson, P.</dc:creator>
<dc:creator>Levi, D.</dc:creator>
<dc:creator>Molina, M.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Downward, J.</dc:creator>
<dc:date>2021-02-02</dc:date>
<dc:identifier>doi:10.1101/2021.02.02.429358</dc:identifier>
<dc:title><![CDATA[Characterisation of tumour immune microenvironment remodelling following oncogene inhibition in preclinical studies using an optimised imaging mass cytometry workflow]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.01.30.428948v1?rss=1">
<title>
<![CDATA[
Mixology: a tool for calculating required masses and volumes for laboratory solutions 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.01.30.428948v1?rss=1"
</link>
<description><![CDATA[
We have created a tool to calculate the amount of each ingredient required to make up a custom solution with defined concentrations. It can convert between many kinds of volumetric, mass and concentration units. This includes the ability to convert between molarities and mass-based concentrations, using molecular masses retrieved from the ChEBI database. Mixology can be accessed at http://mixology.science.
]]></description>
<dc:creator>Sanderson, T.</dc:creator>
<dc:date>2021-01-31</dc:date>
<dc:identifier>doi:10.1101/2021.01.30.428948</dc:identifier>
<dc:title><![CDATA[Mixology: a tool for calculating required masses and volumes for laboratory solutions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.02.03.429355v1?rss=1">
<title>
<![CDATA[
Impact of the B.1.1.7 variant on neutralizing monoclonal antibodies recognizing diverse epitopes on SARS-CoV-2 Spike 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.02.03.429355v1?rss=1"
</link>
<description><![CDATA[
The interaction of the SARS-CoV-2 Spike receptor binding domain (RBD) with the ACE2 receptor on host cells is essential for viral entry. RBD is the dominant target for neutralizing antibodies and several neutralizing epitopes on RBD have been molecularly characterized. Analysis of circulating SARS-CoV-2 variants has revealed mutations arising in the RBD, the N-terminal domain (NTD) and S2 subunits of Spike. To fully understand how these mutations affect the antigenicity of Spike, we have isolated and characterized neutralizing antibodies targeting epitopes beyond the already identified RBD epitopes. Using recombinant Spike as a sorting bait, we isolated >100 Spike-reactive monoclonal antibodies from SARS-CoV-2 infected individuals. ~45% showed neutralizing activity of which ~20% were NTD-specific. None of the S2-specific antibodies showed neutralizing activity. Competition ELISA revealed that NTD-specific mAbs formed two distinct groups: the first group was highly potent against infectious virus, whereas the second was less potent and displayed glycan-dependant neutralization activity. Importantly, mutations present in B.1.1.7 Spike frequently conferred resistance to neutralization by the NTD-specific neutralizing antibodies. This work demonstrates that neutralizing antibodies targeting subdominant epitopes need to be considered when investigating antigenic drift in emerging variants.
]]></description>
<dc:creator>Graham, C.</dc:creator>
<dc:creator>Seow, J.</dc:creator>
<dc:creator>Huettner, I.</dc:creator>
<dc:creator>Khan, H.</dc:creator>
<dc:creator>Kouphou, N.</dc:creator>
<dc:creator>Acors, S.</dc:creator>
<dc:creator>Winstone, H.</dc:creator>
<dc:creator>Pickering, S.</dc:creator>
<dc:creator>Pedro Galao, R.</dc:creator>
<dc:creator>Jose Lista, M.</dc:creator>
<dc:creator>Jimenez-Guardeno, J. M.</dc:creator>
<dc:creator>Laing, A. G.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Joseph, M.</dc:creator>
<dc:creator>Muir, L.</dc:creator>
<dc:creator>Ng, W. M.</dc:creator>
<dc:creator>Duyvesteyn, H. M. E.</dc:creator>
<dc:creator>Zhao, Y.</dc:creator>
<dc:creator>Bowden, T. A.</dc:creator>
<dc:creator>Shankar-Hari, M.</dc:creator>
<dc:creator>Rosa, A.</dc:creator>
<dc:creator>Cherepanov, P.</dc:creator>
<dc:creator>McCoy, L. E.</dc:creator>
<dc:creator>Hayday, A. C.</dc:creator>
<dc:creator>Neil, S. J. D.</dc:creator>
<dc:creator>Malim, M. H.</dc:creator>
<dc:creator>Doores, K. J.</dc:creator>
<dc:date>2021-02-03</dc:date>
<dc:identifier>doi:10.1101/2021.02.03.429355</dc:identifier>
<dc:title><![CDATA[Impact of the B.1.1.7 variant on neutralizing monoclonal antibodies recognizing diverse epitopes on SARS-CoV-2 Spike]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.02.04.429728v1?rss=1">
<title>
<![CDATA[
An extracellular redox signal triggers calcium release and impacts the asexual development of Toxoplasma gondii 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.02.04.429728v1?rss=1"
</link>
<description><![CDATA[
The ability of an organism to sense and respond to environmental redox fluctuations relies on a signaling network that is incompletely understood in apicomplexan parasites such as Toxoplasma gondii. The impact of changes in redox upon the development of this intracellular parasite is not known. Here, we provide a revised collection of 58 genes containing domains related to canonical antioxidant function, with their encoded proteins widely dispersed throughout different cellular compartments. We demonstrate that addition of exogenous H2O2 to human fibroblasts infected with T. gondii triggers a Ca2+ flux in the cytosol of intracellular parasites that can induce egress. In line with existing models, egress triggered by exogenous H2O2 is reliant upon both Calcium-Dependent Protein Kinase 3 and diacylglycerol kinases. Finally, we show that the overexpression a glutaredoxin-roGFP2 redox sensor fusion protein in the parasitophorous vacuole severely impacts parasite replication. These data highlight the rich redox network that exists in T. gondii, evidencing a link between extracellular redox and intracellular Ca2+ signaling that can culminate in parasite egress. Our findings also indicate that the redox potential of the intracellular environment contributes to normal parasite growth. Combined, our findings highlight the important role of redox as an unexplored regulator of parasite biology.
]]></description>
<dc:creator>Alves, E.</dc:creator>
<dc:creator>Benns, H. J.</dc:creator>
<dc:creator>Magnus, L.</dc:creator>
<dc:creator>Dominicus, C.</dc:creator>
<dc:creator>Dobai, T.</dc:creator>
<dc:creator>Blight, J.</dc:creator>
<dc:creator>Wincott, C. J.</dc:creator>
<dc:creator>Child, M. A.</dc:creator>
<dc:date>2021-02-04</dc:date>
<dc:identifier>doi:10.1101/2021.02.04.429728</dc:identifier>
<dc:title><![CDATA[An extracellular redox signal triggers calcium release and impacts the asexual development of Toxoplasma gondii]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.02.05.429998v1?rss=1">
<title>
<![CDATA[
Analysis of independent cohorts of outbred CFW mice reveals novel loci for behavioral and physiological traits and identifies factors determining reproducibility 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.02.05.429998v1?rss=1"
</link>
<description><![CDATA[
Combining samples for genetic association is standard practice in human genetic analysis of complex traits, but is rarely undertaken in rodent genetics. Here, using 23 phenotypes and genotypes from two independent laboratories, we obtained a sample size of 3,076 commercially available outbred mice and identified 70 loci, more than double the number of loci identified in the component studies. Fine-mapping in the combined sample reduced the number of likely causal variants, with a median reduction in set size of 51%, and indicated novel gene associations, including Pnpo, Ttll6 and GM11545 with bone mineral density, and Psmb9 with weight. However replication at a nominal threshold of 0.05 between the two component studies was surprisingly low, with less than a third of loci identified in one study replicated in the second. In addition to overestimates in the effect size in the discovery sample (Winners Curse), we also found that heterogeneity between studies explained the poor replication, but the contribution of these two factors varied among traits. Available methods to control Winners Curse were contingent on the power of the discovery sample, and depending on the method used, both overestimated and underestimated the true effect. Leveraging these observations we integrated information about replication rates, confounding, and Winners Curse corrected estimates of power to assign variants to one of four confidence levels. Our approach addresses concerns about reproducibility, and demonstrates how to obtain robust results from mapping complex traits in any genome-wide association study.
]]></description>
<dc:creator>Zou, J.</dc:creator>
<dc:creator>Gopalakrishnan, S.</dc:creator>
<dc:creator>Parker, C. C.</dc:creator>
<dc:creator>Nicod, J.</dc:creator>
<dc:creator>Mott, R.</dc:creator>
<dc:creator>Cai, N.</dc:creator>
<dc:creator>Lionikas, A.</dc:creator>
<dc:creator>Davies, R. W.</dc:creator>
<dc:creator>Palmer, A. A.</dc:creator>
<dc:creator>Flint, J.</dc:creator>
<dc:date>2021-02-07</dc:date>
<dc:identifier>doi:10.1101/2021.02.05.429998</dc:identifier>
<dc:title><![CDATA[Analysis of independent cohorts of outbred CFW mice reveals novel loci for behavioral and physiological traits and identifies factors determining reproducibility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.02.06.428911v1?rss=1">
<title>
<![CDATA[
Integrative genome-wide analysis reveals EIF3A as a key downstream regulator of translational repressor protein Musashi 2 (MSI2) 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.02.06.428911v1?rss=1"
</link>
<description><![CDATA[
Musashi 2 (MSI2) is an RNA binding protein (RBP) that regulates asymmetric cell division and cell fate decisions in normal and cancer stem cells. MSI2 appears to repress translation by binding to 3 untranslated regions (3UTRs) of mRNA, but the identity of functional targets remains unknown. Here we used iCLIP to identify direct RNA binding partners of MSI2 and integrated these data with polysome profiling to obtain insights into MSI2 function. iCLIP revealed specific MSI2 binding to thousands of target mRNAs largely in 3UTRs, but translational differences were restricted to a small fraction of these transcripts, indicating that MSI2 regulation is not triggered by simple binding. Instead, the functional targets identified here were bound at higher density and contain more "U/TAG" motifs compared to targets bound non-productively. To further distinguish direct and indirect targets, MSI2 was acutely depleted. Surprisingly, only 50 transcripts were found to undergo translational induction on acute MSI2 loss. Eukaryotic elongation factor 3A (EIF3A) was determined to be an immediate, direct target. We propose that MSI2 down-regulation of EIF3A amplifies these effects on the proteome. Our results also underscore the challenges in defining functional targets of RBP since mere binding does not imply a discernible functional interaction.
]]></description>
<dc:creator>Karmakar, S.</dc:creator>
<dc:creator>Ramirez, O.</dc:creator>
<dc:creator>Paul, K. V.</dc:creator>
<dc:creator>Gupta, A. K.</dc:creator>
<dc:creator>Botti, V.</dc:creator>
<dc:creator>Mozos, I. R. d. l.</dc:creator>
<dc:creator>Neuenkirchen, N.</dc:creator>
<dc:creator>Ross, R. J.</dc:creator>
<dc:creator>Neugebauer, K. M.</dc:creator>
<dc:creator>Pillai, M. M.</dc:creator>
<dc:date>2021-02-07</dc:date>
<dc:identifier>doi:10.1101/2021.02.06.428911</dc:identifier>
<dc:title><![CDATA[Integrative genome-wide analysis reveals EIF3A as a key downstream regulator of translational repressor protein Musashi 2 (MSI2)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.02.11.430828v1?rss=1">
<title>
<![CDATA[
Comprehensive phenotypic analysis of the Dp1Tyb mouse strain reveals a broad range of Down Syndrome-related phenotypes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.02.11.430828v1?rss=1"
</link>
<description><![CDATA[
Down syndrome (DS), trisomy 21, results in many complex phenotypes including cognitive deficits, heart defects and craniofacial alterations. Phenotypes arise from an extra copy of human chromosome 21 (Hsa21) genes. However, causative genes remain mostly unknown. Animal models enable identification of these genes and pathological mechanisms. The Dp1Tyb mouse model of DS has an extra copy of 63% of Hsa21-orthologous mouse genes. Here, we comprehensively phenotype Dp1Tyb mice and find wide-ranging DS-like phenotypes including aberrant megakaryopoiesis, reduced bone density, and deficits in memory, locomotion, hearing and sleep. Thus, Dp1Tyb mice are an excellent model for studies of many complex DS phenotypes.
]]></description>
<dc:creator>Lana-Elola, E.</dc:creator>
<dc:creator>Cater, H.</dc:creator>
<dc:creator>Watson-Scales, S.</dc:creator>
<dc:creator>Greenaway, S.</dc:creator>
<dc:creator>Mueller-Winkler, J.</dc:creator>
<dc:creator>Gibbins, D.</dc:creator>
<dc:creator>Nemes, M.</dc:creator>
<dc:creator>Slender, A.</dc:creator>
<dc:creator>Hough, T.</dc:creator>
<dc:creator>Keskivali-Bond, P.</dc:creator>
<dc:creator>Scudamore, C. L.</dc:creator>
<dc:creator>Herbert, E.</dc:creator>
<dc:creator>Banks, G. T.</dc:creator>
<dc:creator>Mobbs, H.</dc:creator>
<dc:creator>Canonica, T.</dc:creator>
<dc:creator>Tosh, J.</dc:creator>
<dc:creator>Noy, S.</dc:creator>
<dc:creator>Llorian, M.</dc:creator>
<dc:creator>Nolan, P. M.</dc:creator>
<dc:creator>Griffin, J. L.</dc:creator>
<dc:creator>Good, M.</dc:creator>
<dc:creator>Simon, M.</dc:creator>
<dc:creator>Mallon, A.-M.</dc:creator>
<dc:creator>Wells, S.</dc:creator>
<dc:creator>Fisher, E. M.</dc:creator>
<dc:creator>Tybulewicz, V. L.</dc:creator>
<dc:date>2021-02-11</dc:date>
<dc:identifier>doi:10.1101/2021.02.11.430828</dc:identifier>
<dc:title><![CDATA[Comprehensive phenotypic analysis of the Dp1Tyb mouse strain reveals a broad range of Down Syndrome-related phenotypes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.02.14.431132v1?rss=1">
<title>
<![CDATA[
A Brownian ratchet model for DNA loop extrusion by the cohesin complex 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.02.14.431132v1?rss=1"
</link>
<description><![CDATA[
The cohesin complex topologically encircles DNA to promote sister chromatid cohesion. Alternatively cohesin extrudes DNA loops, thought to reflect chromatin domain formation. Here, we propose a structure-based model explaining both activities, supported by biochemical experiments. ATP and DNA binding to cohesin promote conformational changes that guide DNA through a kleisin gate into a DNA gripping state. Two HEAT-repeat DNA binding modules, associated with cohesins heads and hinge, are now juxtaposed. ATP hydrolysis disassembles the gripping state, allowing unidirectional hinge module movement to complete topological DNA entry. Without initial kleisin gate passage, biased hinge module motion during gripping state resolution creates a Brownian ratchet that drives loop extrusion. Molecular-mechanical simulations of gripping state formation and resolution cycles recapitulate experimentally observed DNA loop extrusion characteristics. Our model extends to asymmetric and symmetric loop extrusion, as well as z-loop formation. Loop extrusion by biased Brownian fluctuations has important implications for chromosomal cohesin function.
]]></description>
<dc:creator>Higashi, T. L.</dc:creator>
<dc:creator>Tang, M.</dc:creator>
<dc:creator>Pobegalov, G.</dc:creator>
<dc:creator>Uhlmann, F.</dc:creator>
<dc:creator>Molodtsov, M.</dc:creator>
<dc:date>2021-02-14</dc:date>
<dc:identifier>doi:10.1101/2021.02.14.431132</dc:identifier>
<dc:title><![CDATA[A Brownian ratchet model for DNA loop extrusion by the cohesin complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.02.15.431291v1?rss=1">
<title>
<![CDATA[
Favourable antibody responses to human coronaviruses in children and adolescents with autoimmune rheumatic diseases 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.02.15.431291v1?rss=1"
</link>
<description><![CDATA[
Differences in humoral immunity to coronaviruses, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), between children and adults remain unexplained and the impact of underlying immune dysfunction or suppression unknown. Here, we examined the antibody immune competence of children and adolescents with prevalent inflammatory rheumatic diseases, juvenile idiopathic arthritis (JIA), juvenile dermatomyositis (JDM) and juvenile systemic lupus erythematosus (JSLE), against the seasonal human coronavirus (HCoV)-OC43 that frequently infects this age group. Despite immune dysfunction and immunosuppressive treatment, JIA, JDM and JSLE patients mounted comparable or stronger responses than healthier peers, dominated by IgG antibodies to HCoV-OC43 spike, and harboured IgG antibodies that cross-reacted with SARS-CoV-2 spike. In contrast, responses to HCoV-OC43 and SARS-CoV-2 nucleoproteins exhibited delayed age-dependent class-switching and were not elevated in JIA, JDM and JSLE patients, arguing against increased exposure. Consequently, autoimmune rheumatic diseases and their treatment were associated with a favourable ratio of spike to nucleoprotein antibodies.
]]></description>
<dc:creator>Deakin, C.</dc:creator>
<dc:creator>Cornish, G.</dc:creator>
<dc:creator>Ng, K.</dc:creator>
<dc:creator>Faulkner, N.</dc:creator>
<dc:creator>Bolland, W.</dc:creator>
<dc:creator>Panova, V.</dc:creator>
<dc:creator>Hope, J.</dc:creator>
<dc:creator>Rosa, A.</dc:creator>
<dc:creator>Harvey, R.</dc:creator>
<dc:creator>Hussain, S.</dc:creator>
<dc:creator>Earl, C.</dc:creator>
<dc:creator>Jebson, B.</dc:creator>
<dc:creator>Wilkinson, M.</dc:creator>
<dc:creator>Marshall, L.</dc:creator>
<dc:creator>O'Brien, K.</dc:creator>
<dc:creator>Rosser, L.</dc:creator>
<dc:creator>Radziszewska, A.</dc:creator>
<dc:creator>Peckham, H.</dc:creator>
<dc:creator>Heaney, J.</dc:creator>
<dc:creator>Rickman, H.</dc:creator>
<dc:creator>Paraskevopoulou, S.</dc:creator>
<dc:creator>Houlihan, C.</dc:creator>
<dc:creator>Spyer, M.</dc:creator>
<dc:creator>Gamblin, S.</dc:creator>
<dc:creator>Mccauley, J.</dc:creator>
<dc:creator>Nastouli, E.</dc:creator>
<dc:creator>Cherepanov, P.</dc:creator>
<dc:creator>Ciurtin, C.</dc:creator>
<dc:creator>Wedderburn, L.</dc:creator>
<dc:creator>Kassiotis, G.</dc:creator>
<dc:date>2021-02-16</dc:date>
<dc:identifier>doi:10.1101/2021.02.15.431291</dc:identifier>
<dc:title><![CDATA[Favourable antibody responses to human coronaviruses in children and adolescents with autoimmune rheumatic diseases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.02.15.431215v1?rss=1">
<title>
<![CDATA[
Characterization of humoral and SARS-CoV-2 specific T cell responses in people living with HIV 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.02.15.431215v1?rss=1"
</link>
<description><![CDATA[
There is an urgent need to understand the nature of immune responses generated against SARS-CoV-2, to better inform risk-mitigation strategies for people living with HIV (PLWH). Although not all PLWH are considered immunosuppressed, residual cellular immune deficiency and ongoing inflammation could influence COVID-19 disease severity, the evolution and durability of protective memory responses. Here, we performed an integrated analysis, characterizing the nature, breadth and magnitude of SARS-CoV-2-specific immune responses in PLWH, controlled on ART, and HIV negative subjects. Both groups were in the convalescent phase of predominately mild COVID-19 disease. The majority of PLWH mounted SARS-CoV-2 Spike- and Nucleoprotein-specific antibodies with neutralizing activity and SARS-CoV-2-specific T cell responses, as measured by ELISpot, at levels comparable to HIV negative subjects. T cell responses against Spike, Membrane and Nucleocapsid were the most prominent, with SARS-CoV-2-specific CD4 T cells outnumbering CD8 T cells. Notably, the overall magnitude of SARS-CoV-2-specific T cell responses related to the size of the naive CD4 T cell pool and the CD4:CD8 ratio in PLWH, in whom disparate antibody and T cell responses were observed. Both humoral and cellular responses to SARS-CoV-2 were detected at 5-7 months post-infection, providing evidence of medium-term durability of responses irrespective of HIV serostatus. Incomplete immune reconstitution on ART and a low CD4:CD8 ratio could, however, hamper the development of immunity to SARS-CoV-2 and serve as a useful tool for risk stratification of PLWH. These findings have implications for the individual management and potential effectiveness of vaccination against SARS-CoV-2 in PLWH.

One Sentence SummaryAdaptive immune responses to SARS-CoV-2 in the setting of HIV infection
]]></description>
<dc:creator>Alrubayyi, A.</dc:creator>
<dc:creator>Gea-Mallorqui, E.</dc:creator>
<dc:creator>Touizer, E.</dc:creator>
<dc:creator>Hameiri-Bowen, D.</dc:creator>
<dc:creator>Kopycinski, J.</dc:creator>
<dc:creator>Charlton, B.</dc:creator>
<dc:creator>Fisher-Pearson, N.</dc:creator>
<dc:creator>Muir, L.</dc:creator>
<dc:creator>Rosa, A.</dc:creator>
<dc:creator>Roustan, C.</dc:creator>
<dc:creator>Earl, C.</dc:creator>
<dc:creator>Cherepanov, P.</dc:creator>
<dc:creator>Pellegrino, P.</dc:creator>
<dc:creator>Waters, L.</dc:creator>
<dc:creator>Burns, F.</dc:creator>
<dc:creator>Kinloch, S.</dc:creator>
<dc:creator>Dong, T.</dc:creator>
<dc:creator>Dorrell, L.</dc:creator>
<dc:creator>Rowland-Jones, S.</dc:creator>
<dc:creator>McCoy, L. E.</dc:creator>
<dc:creator>Peppa, D.</dc:creator>
<dc:date>2021-02-16</dc:date>
<dc:identifier>doi:10.1101/2021.02.15.431215</dc:identifier>
<dc:title><![CDATA[Characterization of humoral and SARS-CoV-2 specific T cell responses in people living with HIV]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.02.17.431573v1?rss=1">
<title>
<![CDATA[
Inferring population histories for ancient genomes using genome-wide genealogies 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.02.17.431573v1?rss=1"
</link>
<description><![CDATA[
Ancient genomes anchor genealogies in directly observed historical genetic variation, and contextualise ancestral lineages with archaeological insights into their geography and lifestyles. We introduce an extension of the Relate algorithm to incorporate ancient genomes and reconstruct the joint genealogies of 14 previously published high-coverage ancients and 278 present-day individuals of the Simons Genome Diversity Project. As the majority of ancient genomes are of lower coverage and cannot be directly built into genealogies, we additionally present a fast and scalable method, Colate, for inferring coalescence rates between low-coverage genomes without requiring phasing or imputation. Our method leverages sharing patterns of mutations dated using a genealogy to construct a likelihood, which is maximised using an expectation-maximisation algorithm. We apply Colate to 430 ancient human shotgun genomes of >0.5x mean coverage. Using Relate and Colate, we characterise dynamic population structure, such as repeated partial population replacements in Ireland, and gene-flow between early farmer and European hunter-gatherer groups. We further show that the previously reported increase in the TCC/TTC mutation rate, which is strongest in West Eurasians among present-day people, was already widespread across West Eurasia in the Late Glacial Period ~10k - 15k years ago, is strongest in Neolithic and Anatolian farmers, and is remarkably well predicted by the coalescence rates between other genomes and a 10,000-year-old Anatolian individual. This suggests that the driver of this signal originated in ancestors of ancient Anatolia >14k years ago, but was already absent by the Mesolithic and may indicate a genetic link between the Near East and European hunter-gatherer groups in the Late Paleolithic.
]]></description>
<dc:creator>Speidel, L.</dc:creator>
<dc:creator>Cassidy, L.</dc:creator>
<dc:creator>Davies, R. W.</dc:creator>
<dc:creator>Hellenthal, G.</dc:creator>
<dc:creator>Skoglund, P.</dc:creator>
<dc:creator>Myers, S.</dc:creator>
<dc:date>2021-02-17</dc:date>
<dc:identifier>doi:10.1101/2021.02.17.431573</dc:identifier>
<dc:title><![CDATA[Inferring population histories for ancient genomes using genome-wide genealogies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.02.18.431818v1?rss=1">
<title>
<![CDATA[
Rotavirus susceptibility of antibiotic-treated mice ascribed to diminished expression of interleukin-22 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.02.18.431818v1?rss=1"
</link>
<description><![CDATA[
The commensal microbiota regulates susceptibility to enteric pathogens by fine-tuning mucosal innate immune responses, but how susceptibility to enteric viruses is shaped by the microbiota remains incompletely understood. Past reports have indicated that commensal bacteria may either promote or repress rotavirus replication in the small intestine of mice. We now report that rotavirus replicated more efficiently in the intestines of germ-free and antibiotic-treated mice compared to animals with an unmodified microbiota. Antibiotic treatment also facilitated rotavirus replication in type I and type III interferon (IFN) receptor-deficient mice, revealing IFN-independent proviral effects. Expression of interleukin-22 (IL-22) was strongly diminished in the intestine of antibiotic-treated mice. Treatment with exogenous IL-22 blocked rotavirus replication in microbiota-depleted wild-type and Stat1-/- mice, demonstrating that the antiviral effect of IL-22 in animals with altered microbiome is not dependent on IFN signaling. In antibiotic-treated animals, IL-22-induced a specific set of genes including Fut2, encoding fucosyl-transferase 2 that participates in the biosynthesis of fucosylated glycans which can mediate rotavirus binding. Interestingly, IL-22 also blocked rotavirus replication in antibiotic-treated Fut2-/- mice. Furthermore, IL-22 inhibited rotavirus replication in antibiotic-treated mice lacking key molecules of the necroptosis or pyroptosis pathways of programmed cell death. Taken together, our results demonstrate that IL-22 determines rotavirus susceptibility of antibiotic-treated mice, yet the IL-22-induced effector molecules conferring rotavirus resistance remain elusive.
]]></description>
<dc:creator>Schnepf, D.</dc:creator>
<dc:creator>Hernandez, P.</dc:creator>
<dc:creator>Mahlakoiv, T.</dc:creator>
<dc:creator>Crotta, S.</dc:creator>
<dc:creator>Sullender, M. E.</dc:creator>
<dc:creator>Peterson, S. T.</dc:creator>
<dc:creator>Ohnemus, A.</dc:creator>
<dc:creator>Michiels, C.</dc:creator>
<dc:creator>Gentle, I.</dc:creator>
<dc:creator>Dumoutier, L.</dc:creator>
<dc:creator>Reis, C. A.</dc:creator>
<dc:creator>Diefenbach, A.</dc:creator>
<dc:creator>Wack, A.</dc:creator>
<dc:creator>Baldridge, M. T.</dc:creator>
<dc:creator>Staeheli, P.</dc:creator>
<dc:date>2021-02-18</dc:date>
<dc:identifier>doi:10.1101/2021.02.18.431818</dc:identifier>
<dc:title><![CDATA[Rotavirus susceptibility of antibiotic-treated mice ascribed to diminished expression of interleukin-22]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.02.24.432780v1?rss=1">
<title>
<![CDATA[
Parallel evolution of a splicing program controlling neuronal excitability in flies and mammals 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.02.24.432780v1?rss=1"
</link>
<description><![CDATA[
Neurons draw on alternative splicing for their increased transcriptomic complexity throughout animal phylogeny. To delve into the mechanisms controlling the assembly and evolution of this regulatory layer, we characterized the neuronal microexon program in Drosophila and compared it with that of mammals. We found that in Drosophila, this splicing program is restricted to neurons by the post-transcriptional processing of the enhancer of microexons (eMIC) domain in Srrm234 by Elav and Fne. eMIC deficiency or misexpression leads to widespread neurological alterations largely emerging from impaired neuronal activity, as revealed by a combination of neuronal imaging experiments and cell-type-specific rescues. These defects are associated with the genome-wide skipping of short neural exons, which are strongly enriched in ion channels. Remarkably, we found no overlap of eMIC-regulated exons between flies and mice, illustrating how ancient post-transcriptional programs can evolve independently in different phyla to impact distinct cellular modules while maintaining cell-type specificity.
]]></description>
<dc:creator>Torres-Mendez, A.</dc:creator>
<dc:creator>Pop, S.</dc:creator>
<dc:creator>Bonnal, S.</dc:creator>
<dc:creator>Almudi, I.</dc:creator>
<dc:creator>Roberts, R. J. V.</dc:creator>
<dc:creator>Paolantoni, C.</dc:creator>
<dc:creator>Alcaina, A.</dc:creator>
<dc:creator>Avola, A.</dc:creator>
<dc:creator>Martin Anduaga, A.</dc:creator>
<dc:creator>Haussmann, I. H.</dc:creator>
<dc:creator>Morin, V.</dc:creator>
<dc:creator>Soller, M.</dc:creator>
<dc:creator>Kadener, S.</dc:creator>
<dc:creator>Roignant, J.-Y.</dc:creator>
<dc:creator>Prieto-Godino, L.</dc:creator>
<dc:creator>Irimia, M.</dc:creator>
<dc:date>2021-02-25</dc:date>
<dc:identifier>doi:10.1101/2021.02.24.432780</dc:identifier>
<dc:title><![CDATA[Parallel evolution of a splicing program controlling neuronal excitability in flies and mammals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.02.28.433227v1?rss=1">
<title>
<![CDATA[
Whole-genome doubling-aware copy number phylogenies for cancer evolution with MEDICC2 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.02.28.433227v1?rss=1"
</link>
<description><![CDATA[
Chromosomal instability (CIN) and somatic copy-number alterations (SCNA) play a key role in the evolutionary process that shapes cancer genomes. SC-NAs comprise many classes of clinically relevant events, such as localised amplifications, gains, losses, loss-of-heterozygosity (LOH) events, and recently discovered parallel evolutionary events revealed by multi-sample phasing. These events frequently appear jointly with whole genome doubling (WGD), a transformative event in tumour evolution involving tetraploidization of genomes preceded or followed by individual chromosomal copy-number changes and associated with an overall increase in structural CIN.

While SCNAs have been leveraged for phylogeny reconstruction in the past, existing methods do not take WGD events into account and cannot model parallel evolution. They frequently make use of the infinite sites assumption, do not model horizontal dependencies between adjacent genomic loci and can not infer ancestral genomes. Here we present MEDICC2, a new phylogeny inference algorithm for allele-specific SCNA data that addresses these shortcomings. MEDICC2 dispenses with the infinite sites assumption, models parallel evolution and accurately identifies clonal and subclonal WGD events. It times SCNAs relative to each other, quantifies SCNA burden in single-sample studies and infers phylogenetic trees and ancestral genomes in multi-sample or single-cell sequencing scenarios with thousands of cells.

We demonstrate MEDICC2s ability on simulated data, real-world data of 2,778 single sample tumours from the Pan-cancer analysis of whole genomes (PCAWG), 10 bulk multi-region prostate cancer patients and two recent single-cell datasets of triple-negative breast cancer comprising several thousands of single cells.
]]></description>
<dc:creator>Petkovic, M.</dc:creator>
<dc:creator>Watkins, T. B.</dc:creator>
<dc:creator>Colliver, E. C.</dc:creator>
<dc:creator>Laskina, S.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Haase, K.</dc:creator>
<dc:creator>Schwarz, R. F.</dc:creator>
<dc:date>2021-02-28</dc:date>
<dc:identifier>doi:10.1101/2021.02.28.433227</dc:identifier>
<dc:title><![CDATA[Whole-genome doubling-aware copy number phylogenies for cancer evolution with MEDICC2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.03.01.433314v1?rss=1">
<title>
<![CDATA[
Reduced antibody cross-reactivity following infection with B.1.1.7 than with parental SARS-CoV-2 strains 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.01.433314v1?rss=1"
</link>
<description><![CDATA[
We examined the immunogenicity of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variant B.1.1.7 that arose in the United Kingdom and spread globally. Antibodies elicited by B.1.1.7 infection exhibited significantly reduced recognition and neutralisation of parental strains or of the South Africa B.1.351 variant, than of the infecting variant. The drop in cross-reactivity was more pronounced following B.1.1.7 than parental strain infection, indicating asymmetric heterotypic immunity induced by SARS-CoV-2 variants.
]]></description>
<dc:creator>Faulkner, N.</dc:creator>
<dc:creator>Ng, K.</dc:creator>
<dc:creator>Wu, M.</dc:creator>
<dc:creator>Harvey, R.</dc:creator>
<dc:creator>Hussain, S.</dc:creator>
<dc:creator>Greco, M.</dc:creator>
<dc:creator>Bolland, W.</dc:creator>
<dc:creator>Warchal, S.</dc:creator>
<dc:creator>Margaritis, M.</dc:creator>
<dc:creator>Paraskevopoulou, S.</dc:creator>
<dc:creator>Heaney, J.</dc:creator>
<dc:creator>Rickman, H.</dc:creator>
<dc:creator>Houlihan, C.</dc:creator>
<dc:creator>Spyer, M.</dc:creator>
<dc:creator>Frampton, D.</dc:creator>
<dc:creator>Byott, M.</dc:creator>
<dc:creator>de Oliveira, T.</dc:creator>
<dc:creator>Sigal, A.</dc:creator>
<dc:creator>Kjaer, S.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Beale, R.</dc:creator>
<dc:creator>Gamblin, S.</dc:creator>
<dc:creator>Mccauley, J.</dc:creator>
<dc:creator>Daniels, R.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>Bauer, D.</dc:creator>
<dc:creator>Nastouli, E.</dc:creator>
<dc:creator>Kassiotis, G.</dc:creator>
<dc:date>2021-03-01</dc:date>
<dc:identifier>doi:10.1101/2021.03.01.433314</dc:identifier>
<dc:title><![CDATA[Reduced antibody cross-reactivity following infection with B.1.1.7 than with parental SARS-CoV-2 strains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.03.02.433398v1?rss=1">
<title>
<![CDATA[
Genome-wide screens identify calcium signaling as a key regulator of IgE+ plasma celldifferentiation and survival 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.02.433398v1?rss=1"
</link>
<description><![CDATA[
IgE antibodies protect against toxins and parasites, however, they also mediate allergic reactions. In contrast to other antibody isotypes, B cells switched to IgE respond transiently and do not give rise to long-lived plasma cells (PCs) or memory B cells. Although the intrinsic differences of IgE+ B cells have been linked to signaling by the IgE-B cell receptor (BCR), the molecular pathways controlling their behavior remain poorly understood. Here we employ whole-genome CRISPR screening to identify genes regulating IgE+ B cell proliferation, survival and differentiation into PCs. We show that IgE+ B cells are selectively suppressed by the IgE-BCR signaling to intracellular calcium, which inhibits PC differentiation and limits their lifespan after differentiation. Consequently, manipulation of calcium signaling in vivo enhances IgE+ PC responses. Insights from this pathway shed new light on the self-limiting character of IgE responses and open new avenues to eliminate IgE+ PCs in allergy.
]]></description>
<dc:creator>Newman, R.</dc:creator>
<dc:creator>Tolar, P.</dc:creator>
<dc:date>2021-03-02</dc:date>
<dc:identifier>doi:10.1101/2021.03.02.433398</dc:identifier>
<dc:title><![CDATA[Genome-wide screens identify calcium signaling as a key regulator of IgE+ plasma celldifferentiation and survival]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.03.04.433786v1?rss=1">
<title>
<![CDATA[
Barcode Sequencing and a High-throughput Assay for Chronological Lifespan Uncover Ageing-associated Genes in Fission Yeast 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.04.433786v1?rss=1"
</link>
<description><![CDATA[
Ageing-related processes are largely conserved, with simple organisms remaining the main platform to discover and dissect new ageing-associated genes. Yeasts provide potent model systems to study cellular ageing owing their amenability to systematic functional assays under controlled conditions. Even with yeast cells, however, ageing assays can be laborious and resource-intensive. Here we present improved experimental and computational methods to study chronological lifespan in Schizosaccharomyces pombe. We decoded the barcodes for 3206 mutants of the latest gene-deletion library, enabling the parallel profiling of [~]700 additional mutants compared to previous screens. We then applied a refined method of barcode sequencing (Bar-seq), addressing technical and statistical issues raised by persisting DNA in dead cells and sampling bottlenecks in aged cultures, to screen for mutants showing altered lifespan during stationary phase. This screen identified 341 long-lived mutants and 1246 short-lived mutants which point to many previously unknown ageing-associated genes, including 51 conserved but entirely uncharacterized genes. The ageing-associated genes showed coherent enrichments in processes also associated with human ageing, particularly with respect to ageing in non-proliferative brain cells. We also developed an automated colony-forming unit assay for chronological lifespan to facilitate medium- to high-throughput ageing studies by saving time and resources compared to the traditional assay. Results from the Bar-seq screen showed good agreement with this new assay, validating 33 genes not previously associated with cellular ageing. This study provides an effective methodological platform and identifies many new ageing-associated genes as a framework for analysing cellular ageing in yeast and beyond.
]]></description>
<dc:creator>Romila, C.</dc:creator>
<dc:creator>Townsend, S.</dc:creator>
<dc:creator>Malecki, M.</dc:creator>
<dc:creator>Kamrad, S.</dc:creator>
<dc:creator>Rodriguez-Lopez, M.</dc:creator>
<dc:creator>Hillson, O.</dc:creator>
<dc:creator>Cotobal, C.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:creator>Bahler, J.</dc:creator>
<dc:date>2021-03-04</dc:date>
<dc:identifier>doi:10.1101/2021.03.04.433786</dc:identifier>
<dc:title><![CDATA[Barcode Sequencing and a High-throughput Assay for Chronological Lifespan Uncover Ageing-associated Genes in Fission Yeast]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.03.05.431171v1?rss=1">
<title>
<![CDATA[
Robust optical autofocus system utilizing neural networks trained for extended range and time-course and automated multiwell plate imaging including single molecule localization microscopy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.05.431171v1?rss=1"
</link>
<description><![CDATA[
We present a robust, long-range optical autofocus system for microscopy utilizing machine learning. This can be useful for experiments with long image data acquisition times that may be impacted by defocusing resulting from drift of components, e.g. due to changes in temperature or mechanical drift. It is also useful for automated slide scanning or multiwell plate imaging where the sample(s) to be imaged may not be in the same horizontal plane throughout the image data acquisition. To address the impact of (thermal or mechanical) fluctuations over time in the optical autofocus system itself, we utilise a convolutional neural network (CNN) that is trained over multiple days to account for such fluctuations. To address the trade-off between axial precision and range of the autofocus, we implement orthogonal optical readouts with separate CNN training data, thereby achieving an accuracy well within the 600 nm depth of field of our 1.3 numerical aperture objective lens over a defocus range of up to approximately +/- 100 m. We characterise the performance of this autofocus system and demonstrate its application to automated multiwell plate single molecule localisation microscopy.
]]></description>
<dc:creator>Lightley, J.</dc:creator>
<dc:creator>Görlitz, F.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Kalita, R.</dc:creator>
<dc:creator>Kolbeinsson, A.</dc:creator>
<dc:creator>Garcia, E.</dc:creator>
<dc:creator>Alexandrov, Y.</dc:creator>
<dc:creator>Bousgouni, V.</dc:creator>
<dc:creator>Wysoczanski, R.</dc:creator>
<dc:creator>Barnes, P.</dc:creator>
<dc:creator>Donnelly, L.</dc:creator>
<dc:creator>Bakal, C.</dc:creator>
<dc:creator>Dunsby, C.</dc:creator>
<dc:creator>Neil, M. A. A.</dc:creator>
<dc:creator>Flaxman, S.</dc:creator>
<dc:creator>French, P. M. W.</dc:creator>
<dc:date>2021-03-05</dc:date>
<dc:identifier>doi:10.1101/2021.03.05.431171</dc:identifier>
<dc:title><![CDATA[Robust optical autofocus system utilizing neural networks trained for extended range and time-course and automated multiwell plate imaging including single molecule localization microscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.03.05.434119v1?rss=1">
<title>
<![CDATA[
SARS-CoV-2-host chimeric RNA-sequencing reads do not necessarily signify virus integration into the host DNA 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.05.434119v1?rss=1"
</link>
<description><![CDATA[
The human genome bears evidence of extensive invasion by retroviruses and other retroelements, as well as by diverse RNA and DNA viruses. High frequency of somatic integration of the RNA virus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) into the DNA of infected cells was recently suggested, partly based on the detection of chimeric RNA-sequencing (RNA-seq) reads between SARS-CoV-2 RNA and RNA transcribed from human host DNA. Here, we examined the possible origin of human-SARS-CoV-2 chimeric reads in RNA-seq libraries and provide alternative explanations for their origin. Chimeric reads were frequently detected also between SARS-CoV-2 RNA and RNA transcribed from mitochondrial DNA or episomal adenoviral DNA present in transfected cell lines, which was unlikely the result of SARS-CoV-2 integration. Furthermore, chimeric reads between SARS-CoV-2 RNA and RNA transcribed from nuclear DNA was highly enriched for host exonic, than intronic or intergenic sequences and often involved the same, highly expressed host genes. These findings suggest that human-SARS-CoV-2 chimeric reads found in RNA-seq data may arise during library preparation and do not necessarily signify SARS-CoV-2 reverse transcription, integration in to host DNA and further transcription.
]]></description>
<dc:creator>Kazachenka, A.</dc:creator>
<dc:creator>Kassiotis, G.</dc:creator>
<dc:date>2021-03-05</dc:date>
<dc:identifier>doi:10.1101/2021.03.05.434119</dc:identifier>
<dc:title><![CDATA[SARS-CoV-2-host chimeric RNA-sequencing reads do not necessarily signify virus integration into the host DNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.03.08.434385v1?rss=1">
<title>
<![CDATA[
High sensitivity dia-PASEF proteomics with DIA-NN and FragPipe 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.08.434385v1?rss=1"
</link>
<description><![CDATA[
The dia-PASEF technology exploits ion mobility separation for high-sensitivity analysis of complex proteomes. Here, we demonstrate neural network-based processing of the ion mobility data, which we implement in the DIA-NN software suite. Using spectral libraries generated with the MSFragger-based FragPipe computational platform, the DIA-NN analysis of dia-PASEF raw data increases the proteomic depth by up to 69% compared to the originally published dia-PASEF workflow. For example, we quantify over 5200 proteins from 10ng of HeLa peptides separated with a 95-minute nanoflow gradient, and over 5000 proteins from 200ng using a 4.8-minute separation with an Evosep One system. In complex samples, featuring a mix of human and yeast lysates, the workflow detects over 11700 proteins in single runs acquired with a 100-minute nanoflow gradient, while demonstrating quantitative precision. Hence, the combination of FragPipe and DIA-NN provides a simple-to-use software platform for dia-PASEF data analysis, yielding significant gains in high-sensitivity proteomics.
]]></description>
<dc:creator>Demichev, V.</dc:creator>
<dc:creator>Yu, F.</dc:creator>
<dc:creator>Teo, G. C.</dc:creator>
<dc:creator>Szyrwiel, L.</dc:creator>
<dc:creator>Rosenberger, G.</dc:creator>
<dc:creator>Decker, J.</dc:creator>
<dc:creator>Kaspar-Schoenefeld, S.</dc:creator>
<dc:creator>Lilley, K. S.</dc:creator>
<dc:creator>Muelleder, M.</dc:creator>
<dc:creator>Nesvizhskii, A. I.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:date>2021-03-09</dc:date>
<dc:identifier>doi:10.1101/2021.03.08.434385</dc:identifier>
<dc:title><![CDATA[High sensitivity dia-PASEF proteomics with DIA-NN and FragPipe]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.09.18.304139v1?rss=1">
<title>
<![CDATA[
Antisense oligonucleotides target a nearly invariant structural element from the SARS-CoV-2 genome and drive RNA degradation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.09.18.304139v1?rss=1"
</link>
<description><![CDATA[
RNA structural elements occur in numerous single stranded (+)-sense RNA viruses. The stemloop 2 motif (s2m) is one such element with an unusually high degree of sequence conservation, being found in the 3 UTR in the genomes of many astroviruses, some picornaviruses and noroviruses, and a variety of coronaviruses, including SARS-CoV and SARS-CoV-2. The evolutionary conservation and its occurrence in all viral subgenomic transcripts implicates a key role of s2m in the viral infection cycle. Our findings indicate that the element, while stably folded, can nonetheless be invaded and remodelled spontaneously by antisense oligonucleotides (ASOs) that initiate pairing in exposed loops and trigger efficient sequence-specific RNA cleavage in reporter assays. ASOs also act to inhibit replication in an astrovirus replicon model system in a sequence-specific, dose-dependent manner and inhibit SARS-CoV-2 infection in cell culture. Our results thus permit us to suggest that the s2m element is a site of vulnerability readily targeted by ASOs, which show promise as anti-viral agents.
]]></description>
<dc:creator>Lulla, V.</dc:creator>
<dc:creator>Wandel, M. P.</dc:creator>
<dc:creator>Bandyra, K. J.</dc:creator>
<dc:creator>Dendooven, T.</dc:creator>
<dc:creator>Yang, X.</dc:creator>
<dc:creator>Doyle, N.</dc:creator>
<dc:creator>Oerum, S.</dc:creator>
<dc:creator>O'Rourke, S.</dc:creator>
<dc:creator>Randow, F.</dc:creator>
<dc:creator>Maier, H. J.</dc:creator>
<dc:creator>Scott, W.</dc:creator>
<dc:creator>Ding, Y.</dc:creator>
<dc:creator>Firth, A.</dc:creator>
<dc:creator>Bloznelyte, K.</dc:creator>
<dc:creator>Luisi, B.</dc:creator>
<dc:date>2020-09-19</dc:date>
<dc:identifier>doi:10.1101/2020.09.18.304139</dc:identifier>
<dc:title><![CDATA[Antisense oligonucleotides target a nearly invariant structural element from the SARS-CoV-2 genome and drive RNA degradation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.03.11.434982v1?rss=1">
<title>
<![CDATA[
A quantitative landscape of cell fate transitions identifies principles of cellular decision-making 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.11.434982v1?rss=1"
</link>
<description><![CDATA[
Fate decisions in developing tissues involve cells transitioning between a set of discrete cell states, each defined by a distinct gene expression profile. Geometric models, often referred to as Waddington landscapes, in which developmental paths are given by the gradient and cell states by the minima of the model, are an appealing way to describe differentiation dynamics and developmental decisions. To construct and validate accurate dynamical landscapes, quantitative methods based on experimental data are necessary. To this end we took advantage of the differentiation of neural and mesodermal cells from pluripotent mouse embryonic stem cells exposed to different combinations and durations of signalling factors. We developed a principled statistical approach using flow cytometry data to quantify differentiating cell states. Then, using a framework based on Catastrophe Theory and approximate Bayesian computation, we constructed the corresponding dynamical landscape. The result was a quantitative model that accurately predicted the proportions of neural and mesodermal cells differentiating in response to specific signalling regimes. Analysis of the geometry of the landscape revealed two distinct ways in which cells make a binary choice between one of two fates. We discuss the biological relevance of these mechanisms and suggest that they represent general archetypal designs for developmental decisions. Taken together, the approach we describe is broadly applicable for the quantitative analysis of differentiation dynamics and for determining the logic of developmental cell fate decisions.
]]></description>
<dc:creator>Saez, M.</dc:creator>
<dc:creator>Blassberg, R.</dc:creator>
<dc:creator>Camacho-Aguilar, E.</dc:creator>
<dc:creator>Siggia, E. D.</dc:creator>
<dc:creator>Rand, D. A.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:date>2021-03-11</dc:date>
<dc:identifier>doi:10.1101/2021.03.11.434982</dc:identifier>
<dc:title><![CDATA[A quantitative landscape of cell fate transitions identifies principles of cellular decision-making]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.03.13.435244v1?rss=1">
<title>
<![CDATA[
Crosstalk between enterocytes and innate lymphoid cells drives early IFN-g-mediated control of Cryptosporidium 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.13.435244v1?rss=1"
</link>
<description><![CDATA[
The intestinal parasite, Cryptosporidium, is a major contributor to global child mortality and causes opportunistic infection in immune deficient individuals. Innate resistance to Cryptosporidium, which specifically invades enterocytes, is dependent on the production of IFN-{gamma}, yet whether enterocytes contribute to parasite control is poorly understood. In this study, utilizing the natural mouse pathogen, Cryptosporidium tyzzeri, we show that epithelial-derived IL-18 synergized with IL-12 to stimulate innate lymphoid cell (ILC) production of IFN-{gamma}. This innate IFN-{gamma} was required for early parasite control. Loss of STAT1 in enterocytes, but not dendritic cells or macrophages, antagonized early parasite control. Transcriptional profiling of enterocytes from infected mice identified an IFN-{gamma} signature and enrichment of anti-microbial effectors like IDO, GBP and IRG. Deletion experiments identified a role for Irgm1/m3 in parasite control. Thus, enterocytes promote ILC production of IFN-{gamma} that acts on enterocytes to restrict the growth of C. tyzzeri.
]]></description>
<dc:creator>Gullicksrud, J.</dc:creator>
<dc:creator>Sateriale, A.</dc:creator>
<dc:creator>Engiles, J. B.</dc:creator>
<dc:creator>Gibson, A.</dc:creator>
<dc:creator>Shaw, S.</dc:creator>
<dc:creator>Hutchins, Z.</dc:creator>
<dc:creator>Martin, L.</dc:creator>
<dc:creator>Christian, D. A.</dc:creator>
<dc:creator>Taylor, G. A.</dc:creator>
<dc:creator>Yamamoto, M.</dc:creator>
<dc:creator>Beiting, D. P.</dc:creator>
<dc:creator>Striepen, B.</dc:creator>
<dc:creator>Hunter, C.</dc:creator>
<dc:date>2021-03-14</dc:date>
<dc:identifier>doi:10.1101/2021.03.13.435244</dc:identifier>
<dc:title><![CDATA[Crosstalk between enterocytes and innate lymphoid cells drives early IFN-g-mediated control of Cryptosporidium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.03.15.434938v1?rss=1">
<title>
<![CDATA[
High Content and High Throughout Phenotypic Assay for the Hourly Resolution of the Malaria Parasite Erythrocytic Cycle 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.15.434938v1?rss=1"
</link>
<description><![CDATA[
Over the last 20 years increased funding for malaria research has resulted in very significant technical advances to study the biology of Plasmodium species. High throughput phenotypic assays have been developed to screen millions of compounds and identify small molecules with antiparasitic activity. At the same time, advances in malaria genetic have greatly facilitated the generation of genetically modified parasites, and whole genome genetic screens are now feasible in Plasmodium species. Finally, there has been an increased interest to study malaria parasites at the population level, in particular in the area of drug resistance. Drug resistant field isolates have been collected around the world, and drug resistant strains are routinely generated in the lab to study the mechanisms of drug resistance. As a result, one of the current bottlenecks in malaria research is our ability to quickly characterize the phenotype associated with compound treatment or genetic modification, or to quickly compare differences in intracellular development between strains. Here, we present a high content/high throughput phenotypic assay that combines highly selective RNA, DNA, and RBC membrane dyes to provide hourly resolution of the full erythrocytic cycle for both P. falciparum and P. knowlesi. A flow cytometry assay allows the analysis of samples in a 384-well format and a quick way to determine the parasite developmental stage. On the other hand, the fluorescence microscopy format allows for a detailed visualization of parasite morphology. Finally, using open source software we have developed protocols for the automated cluster analysis of microscopy images. This assay can be applied to any Plasmodium species, requires very little amount of sample, is performed with fixed cells, and is easily scalable. Overall, we believe this assay will be a great tool for the malaria community to study Plasmodium species.
]]></description>
<dc:creator>Bell, D.</dc:creator>
<dc:creator>Ridewood, S.</dc:creator>
<dc:creator>Patel, A. P.</dc:creator>
<dc:creator>Lee, S. H.</dc:creator>
<dc:creator>Chang, Y.-T.</dc:creator>
<dc:creator>Deu, E.</dc:creator>
<dc:date>2021-03-15</dc:date>
<dc:identifier>doi:10.1101/2021.03.15.434938</dc:identifier>
<dc:title><![CDATA[High Content and High Throughout Phenotypic Assay for the Hourly Resolution of the Malaria Parasite Erythrocytic Cycle]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.03.15.433711v1?rss=1">
<title>
<![CDATA[
Development of novel MOG analogues with increased stability to explore MCT2 and α-ketoglutarate biology in vivo 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.15.433711v1?rss=1"
</link>
<description><![CDATA[
-ketoglutarate (KG) is a central metabolic node with a broad influence on cellular physiology. The KG analogue N-oxalylglycine (NOG) and its membrane-permeable pro-drug derivative dimethyl-oxalylglycine (DMOG) have been extensively used as tools to study prolyl hydroxylases (PHDs) and other KG-dependent processes. In cell culture media, DMOG is rapidly converted to MOG, which enters cells through monocarboxylate transporter MCT2, leading to intracellular NOG concentrations that are sufficiently high to inhibit glutaminolysis enzymes and cause cytotoxicity. Therefore, the degree of (D)MOG instability together with MCT2 expression levels determine the intracellular targets NOG engages with and, ultimately, its effects on cell viability. Here we designed and characterised a series of MOG analogues with the aims of improving stability and exploring the functional requirements for interaction with MCT2, a relatively understudied member of the SLC16 family. We report MOG analogues that maintain ability to enter cells via MCT2, and identify compounds that do not inhibit glutaminolysis or cause cytotoxicity but can still inhibit PHDs. We use these analogues to show that glutaminolysis-induced activation of mTORC1 can be uncoupled from PHD activity. Therefore, these new compounds can help deconvolute cellular effects that result from the polypharmacological action of NOG.
]]></description>
<dc:creator>Fets, L.</dc:creator>
<dc:creator>Nunes, P. M.</dc:creator>
<dc:creator>Campos, S.</dc:creator>
<dc:creator>Silva dos Santos, M.</dc:creator>
<dc:creator>Bevan, N.</dc:creator>
<dc:creator>Sherriff, E. B.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>House, D.</dc:creator>
<dc:creator>Anastasiou, D.</dc:creator>
<dc:date>2021-03-15</dc:date>
<dc:identifier>doi:10.1101/2021.03.15.433711</dc:identifier>
<dc:title><![CDATA[Development of novel MOG analogues with increased stability to explore MCT2 and α-ketoglutarate biology in vivo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.03.15.433780v1?rss=1">
<title>
<![CDATA[
Epiblast morphogenesis is controlled by selective mRNA decay triggered by LIN28A relocation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.15.433780v1?rss=1"
</link>
<description><![CDATA[
The embryonic progression from naive to primed pluripotency is accompanied by the rapid decay of pluripotency-associated mRNAs and a concomitant radical morphogenetic sequence of epiblast polarization, rosette formation and lumenogenesis. The mechanisms triggering and linking these events remain poorly understood. Guided by machine learning and metabolic RNA sequencing, we identified RNA binding proteins (RBPs), especially LIN28A, as primary mRNA decay factors. Using mRNA-RBP interactome capture, we revealed a dramatic increase in LIN28A mRNA binding during the naive-rosette-primed pluripotency transition, driven by its nucleolar-to-cytoplasmic translocation. Cytoplasmic LIN28A binds to 3UTRs of pluripotency-associated mRNAs to directly stimulate their decay and drive lumenogenesis. Accordingly, forced nuclear retention of LIN28A impeded lumenogenesis, impaired gastrulation, and caused an unforeseen embryonic multiplication. Selective mRNA decay, driven by nucleo-cytoplasmic RBP translocation, therefore acts as an intrinsic mechanism linking cell identity switches to the control of embryonic growth and morphogenesis.
]]></description>
<dc:creator>Modic, M.</dc:creator>
<dc:creator>los Mozos, I. R. d.</dc:creator>
<dc:creator>Steinhauser, S.</dc:creator>
<dc:creator>van Genderen, E.</dc:creator>
<dc:creator>Schirge, S.</dc:creator>
<dc:creator>Bergant, V.</dc:creator>
<dc:creator>Ryan, J.</dc:creator>
<dc:creator>Mulholland, C. B.</dc:creator>
<dc:creator>Lee, F. C. Y.</dc:creator>
<dc:creator>Faraway, R.</dc:creator>
<dc:creator>Klobucar, T.</dc:creator>
<dc:creator>Merl-Pham, J.</dc:creator>
<dc:creator>Hauck, S. M.</dc:creator>
<dc:creator>Drukker, M.</dc:creator>
<dc:creator>Bultmann, S.</dc:creator>
<dc:creator>Leonhardt, H.</dc:creator>
<dc:creator>Lickert, H.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:creator>ten Berge, D.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:date>2021-03-16</dc:date>
<dc:identifier>doi:10.1101/2021.03.15.433780</dc:identifier>
<dc:title><![CDATA[Epiblast morphogenesis is controlled by selective mRNA decay triggered by LIN28A relocation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.03.18.436028v1?rss=1">
<title>
<![CDATA[
HIV-1 requires capsid remodelling at the nuclear pore for nuclear entry and integration 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.18.436028v1?rss=1"
</link>
<description><![CDATA[
The capsid (CA) lattice of the HIV-1 core plays a key role during infection. From the moment the core is released into the cytoplasm, it interacts with a range of cellular factors that, ultimately, direct the pre-integration complex to the integration site. For integration to occur, the CA lattice must disassemble. Early uncoating or a failure to do so has detrimental effects on virus infectivity, indicating that an optimal stability of the viral core is crucial for infection. Here, we introduced cysteine residues into HIV-1 CA in order to induce disulphide bond formation and engineer hyper-stable mutants that are slower or unable to uncoat, and then followed their replication. From a panel of mutants, we identified three with increased capsid stability in cells and found that, whilst the M68C/E212C mutant had a 5-fold reduction in reverse transcription, two mutants, A14C/E45C and E180C, were able to reverse transcribe to approximately WT levels. Moreover, these mutants only had a 5-fold reduction in 2-LTR circle production, suggesting that not only could reverse transcription complete in hyper-stable cores, but that the nascent viral cDNA could enter the nuclear compartment. Furthermore, we observed significant levels of A14C/E45C mutant capsid in nuclear and chromatin-associated fractions implying that the hyper-stable cores themselves entered the nucleus. Immunofluorescence studies revealed that although the A14C/E45C mutant capsid reached the nuclear pore with the same kinetics as wild type capsid, it was then retained at the pore in association with Nup153. Crucially, infection with the hyper-stable mutants did not promote CPSF6 re-localisation to nuclear speckles, despite the mutant capsids being competent for CPSF6 binding. These observations suggest that hyper-stable cores are not able to uncoat, or remodel, enough to pass through or dissociate from the nuclear pore and integrate successfully. This, is turn, highlights the importance of capsid lattice flexibility for nuclear entry. In conclusion, we hypothesise that during a productive infection, a capsid remodelling step takes place at the nuclear pore that releases the core complex from Nup153, and relays it to CPSF6, which then localises it to chromatin ready for integration.

AUTHOR SUMMARYThe mature viral core of human immunodeficiency virus (HIV) consists of a highly organised lattice formed by capsid molecules that encloses the viral RNA and viral enzymes. This lattice is crucial during the early stages of viral replication, as it has to break down - uncoat - at the right time and place in order for the viral DNA to integrate successfully. Lentiviruses, like HIV, can infect non-dividing cells and are able to access the host cell DNA by entering the nucleus through nuclear pores. Until recently, uncoating was thought to occur in the cytoplasm as the whole core was thought too large to pass through the nuclear pore. However, lately it has been suggested that uncoating might occur at the nuclear pore or even inside the nucleus and the site of uncoating is currently hotly debated. By investigating HIV mutants with an increased lattice stability, we have shown that lattice flexibility is crucial for nuclear entry. Provocatively, we observed hyper-stable mutant capsid in nuclear and chromatin-associated fractions suggesting that uncoating is not required for nuclear entry. Nonetheless, microscopy experiments suggested that these hyper-stable mutants were retained on the inner side of the nuclear pore, and were impaired for downstream events in the nucleus, leading to a severe infectivity defect. Therefore, we believe that an essential uncoating, or capsid lattice remodelling event normally takes place at the nuclear pore.
]]></description>
<dc:creator>Guedan, A.</dc:creator>
<dc:creator>Donaldson, C. D.</dc:creator>
<dc:creator>Cosnefroy, O.</dc:creator>
<dc:creator>Taylor, I. A.</dc:creator>
<dc:creator>Bishop, K. N.</dc:creator>
<dc:date>2021-03-18</dc:date>
<dc:identifier>doi:10.1101/2021.03.18.436028</dc:identifier>
<dc:title><![CDATA[HIV-1 requires capsid remodelling at the nuclear pore for nuclear entry and integration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.03.20.436222v1?rss=1">
<title>
<![CDATA[
N-myristoyltransferase inhibition is synthetic lethal in MYC-deregulated cancers 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.20.436222v1?rss=1"
</link>
<description><![CDATA[
Human N-myristoyltransferases (NMTs) catalyze N-terminal protein myristoylation, a modification regulating membrane trafficking and interactions of >100 proteins. NMT is a promising target in cancer, but a mechanistic rationale for targeted therapy remains poorly defined. Here, large-scale cancer cell line screens against a panel of NMT inhibitors (NMTi) were combined with systems-level analyses to reveal that NMTi is synthetic lethal with deregulated MYC. Synthetic lethality is mediated by post-transcriptional failure in mitochondrial respiratory complex I protein synthesis concurrent with loss of myristoylation and degradation of complex I assembly factor NDUFAF4, followed by mitochondrial dysfunction specifically in MYC-deregulated cancer cells. NMTi eliminated MYC-deregulated tumors in vivo without overt toxicity, providing a new paradigm in which targeting a constitutive co-translational protein modification is synthetically lethal in MYC-deregulated cancers.

One-sentence summaryN-myristoyltransferase inhibition leads to post-transcriptional complex I failure and cell death in MYC-deregulated cancers
]]></description>
<dc:creator>Lueg, G. A.</dc:creator>
<dc:creator>Faronato, M.</dc:creator>
<dc:creator>Gorelik, A.</dc:creator>
<dc:creator>Goya Grocin, A.</dc:creator>
<dc:creator>Caamano-Gutierrez, E.</dc:creator>
<dc:creator>Falciani, F.</dc:creator>
<dc:creator>Solari, R.</dc:creator>
<dc:creator>Carr, R.</dc:creator>
<dc:creator>Bell, A. S.</dc:creator>
<dc:creator>Bartlett, E.</dc:creator>
<dc:creator>Hutton, J. A.</dc:creator>
<dc:creator>Llorian-Sopena, M.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Brzezicha, B.</dc:creator>
<dc:creator>Janz, M.</dc:creator>
<dc:creator>Garnett, M. J.</dc:creator>
<dc:creator>Calado, D. P.</dc:creator>
<dc:creator>Tate, E. W.</dc:creator>
<dc:date>2021-03-20</dc:date>
<dc:identifier>doi:10.1101/2021.03.20.436222</dc:identifier>
<dc:title><![CDATA[N-myristoyltransferase inhibition is synthetic lethal in MYC-deregulated cancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.03.19.436198v1?rss=1">
<title>
<![CDATA[
ChromWave: Deciphering the DNA-encoded competition between transcription factors and nucleosomes with deep neural networks 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.19.436198v1?rss=1"
</link>
<description><![CDATA[
Transcription factors (TFs) regulate gene expression by recognising and binding specific DNA sequences. At times, these regulatory elements may be occluded by nucleosomes, making them inaccessible for TF-binding. The competition for DNA occupancy between TFs and nucleosomes, and associated gene regulatory outputs, are important consequences of the cis-regulatory information encoded in the genome. However, these sequence patterns are subtle and remain difficult to interpret. Here, we introduce ChromWave, a deep-learning model that, for the first time, predicts the competing profiles for TF and nucleosomes occupancies with remarkable accuracy. Models trained using short- and long-fragment MNase-Seq data successfully learn the sequence preferences underlying TF and nucleosome occupancies across the entire yeast genome. They recapitulate nucleosome evictions from regions containing "strong" TF binding sites and knock-out simulations show nucleosomes gaining occupancy in the absence of these TFs, accompanied by lateral rearrangement of adjacent nucleosomes. At a local level, models anticipate with high accuracy the outcomes of detailed experimental analysis of partially unwrapped nucleosomes at the GAL4 UAS locus. Finally, we trained a ChromWave model that successfully predicts nucleosome positions at promoters in the human genome. We find that human promoters generally contain few sites at which simple sequence changes can alter nucleosome occupancies and that these positions align well with causal variants linked to DNase hypersensitivity.
]]></description>
<dc:creator>Cakiroglu, S. A.</dc:creator>
<dc:creator>Steinhauser, S.</dc:creator>
<dc:creator>Smith, J.</dc:creator>
<dc:creator>Xing, W.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:date>2021-03-20</dc:date>
<dc:identifier>doi:10.1101/2021.03.19.436198</dc:identifier>
<dc:title><![CDATA[ChromWave: Deciphering the DNA-encoded competition between transcription factors and nucleosomes with deep neural networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.03.24.436749v1?rss=1">
<title>
<![CDATA[
TRIM28-dependent SUMOylation protects the adult ovary from the male pathway 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.24.436749v1?rss=1"
</link>
<description><![CDATA[
Gonadal sexual fate in mammals is determined during embryonic development and must be actively maintained in adulthood. In the mouse ovary, oestrogen receptors and FOXL2 protect ovarian granulosa cells from transdifferentiation into Sertoli cells, their testicular counterpart. However, the mechanism underlying their protective effect is unknown. Here, we show that TRIM28 is required to prevent female-to-male sex reversal of the mouse ovary after birth. We found that upon loss of Trim28, ovarian granulosa cells transdifferentiate to Sertoli cells through an intermediate cell type, different from gonadal embryonic progenitors. TRIM28 is recruited on chromatin in the proximity of FOXL2 to maintain the ovarian pathway and to repress testicular-specific genes. The role of TRIM28 in ovarian maintenance depends on its E3-SUMO ligase activity that regulates the sex-specific SUMOylation profile of ovarian-specific genes. Our study identifies TRIM28 as a key factor in protecting the adult ovary from the testicular pathway.
]]></description>
<dc:creator>Rossitto, M.</dc:creator>
<dc:creator>Dejardin, S.</dc:creator>
<dc:creator>Rands, C. M.</dc:creator>
<dc:creator>Legras, S.</dc:creator>
<dc:creator>Migale, R.</dc:creator>
<dc:creator>Rafiee, M.-R.</dc:creator>
<dc:creator>Neirijnck, Y.</dc:creator>
<dc:creator>Pruvost, A.</dc:creator>
<dc:creator>Nguyen, A. L.</dc:creator>
<dc:creator>Bossis, G.</dc:creator>
<dc:creator>Cammas, F. M.</dc:creator>
<dc:creator>Le Gallic, L.</dc:creator>
<dc:creator>Wilhelm, D.</dc:creator>
<dc:creator>Boizet-Bonhoure, B.</dc:creator>
<dc:creator>Lovell-Badge, R.</dc:creator>
<dc:creator>Nef, S.</dc:creator>
<dc:creator>Poulat, F.</dc:creator>
<dc:date>2021-03-24</dc:date>
<dc:identifier>doi:10.1101/2021.03.24.436749</dc:identifier>
<dc:title><![CDATA[TRIM28-dependent SUMOylation protects the adult ovary from the male pathway]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.03.30.437698v1?rss=1">
<title>
<![CDATA[
In vitro teratogenicity testing using a 3D, embryo-like gastruloid system 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.30.437698v1?rss=1"
</link>
<description><![CDATA[
Pharmaceuticals that are intended for use in patients of childbearing potential need to be tested for teratogenicity before marketing. Several pharmaceutical companies use animal-free in vitro models which allow a more rapid selection of lead compounds and contribute to 3Rs principles ( replace, reduce and refine) by streamlining the selection of promising compounds that are submitted to further regulatory studies in animals. Currently available in vitro models typically rely on adherent monolayer cultures or disorganized 3D structures, both of which lack the spatiotemporal and morphological context of the developing embryo. A newly developed 3D  gastruloid model has the potential to achieve a more reliable prediction of teratogenicity by providing a robust recapitulation of gastrulation-like events alongside morphological coordination at relatively high-throughput. In this first proof-of-concept study, we used both mouse and human gastruloids to examine a panel of seven reference compounds, with associated in vivo data and known teratogenic risk, to quantitatively assess in vitro teratogenicity. We observed several gross morphological effects, including significantly reduced elongation or decreased size of the gastruloids, upon exposure to several of the reference compounds. We also observed aberrant gene expression using fluorescent reporters, including SOX2, BRA, and SOX17, suggestive of multi-lineage differentiation defects and disrupted axial patterning. Finally, we saw that gastruloids recapitulated some of the known in vivo species-specific susceptibilities between their mouse and human counterparts. We therefore suggest that gastruloids represent a powerful tool for teratogenicity assessment by enabling relevant physiological recapitulation of early embryonic development, demonstrating their use as a novel in vitro teratogenic model system.
]]></description>
<dc:creator>Mantziou, V.</dc:creator>
<dc:creator>Baillie-Benson, P.</dc:creator>
<dc:creator>Jaklin, M.</dc:creator>
<dc:creator>Kustermann, S.</dc:creator>
<dc:creator>Martinez Arias, A.</dc:creator>
<dc:creator>Moris, N.</dc:creator>
<dc:date>2021-03-30</dc:date>
<dc:identifier>doi:10.1101/2021.03.30.437698</dc:identifier>
<dc:title><![CDATA[In vitro teratogenicity testing using a 3D, embryo-like gastruloid system]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.03.29.437407v1?rss=1">
<title>
<![CDATA[
Biallelic mutations in cancer genomes reveal local mutational determinants 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.29.437407v1?rss=1"
</link>
<description><![CDATA[
The infinite sites model of molecular evolution requires that every position in the genome is mutated at most once1. It is a cornerstone of tumour phylogenetic analysis2, and is often implied when calling, phasing and interpreting variants3,4 or studying the mutational landscape as a whole5. Here we identify 20,555 biallelic mutations, where the same base is mutated independently on both parental copies, in 722 (26.0%) bulk sequencing samples from the Pan-Cancer Analysis of Whole Genomes study (PCAWG). Biallelic mutations reveal UV damage hotspots at ETS and NFAT binding sites, and hypermutable motifs in POLE-mutant and other cancers. We formulate recommendations for variant calling and provide frameworks to model and detect biallelic mutations. These results highlight the need for accurate models of mutation rates and tumour evolution, as well as their inference from sequencing data.
]]></description>
<dc:creator>Demeulemeester, J.</dc:creator>
<dc:creator>Dentro, S. C.</dc:creator>
<dc:creator>Gerstung, M.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:date>2021-03-30</dc:date>
<dc:identifier>doi:10.1101/2021.03.29.437407</dc:identifier>
<dc:title><![CDATA[Biallelic mutations in cancer genomes reveal local mutational determinants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.03.30.437173v1?rss=1">
<title>
<![CDATA[
Severity of SARS-CoV-2 infection as a function of the interferon landscape across the respiratory tract of COVID-19 patients. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.30.437173v1?rss=1"
</link>
<description><![CDATA[
The COVID-19 outbreak driven by SARS-CoV-2 has caused more than 2.5 million deaths globally, with the most severe cases characterized by over-exuberant production of immune-mediators, the nature of which is not fully understood. Interferons of the type I (IFN-I) or type III (IFN-III) families are potent antivirals, but their role in COVID-19 remains debated. Our analysis of gene and protein expression along the respiratory tract shows that IFNs, especially IFN-III, are over-represented in the lower airways of patients with severe COVID-19, while high levels of IFN-III, and to a lesser extent IFN-I, characterize the upper airways of patients with high viral burden but reduced disease risk or severity; also, IFN expression varies with abundance of the cell types that produce them. Our data point to a dynamic process of inter- and intra-family production of IFNs in COVID-19, and suggest that IFNs play opposing roles at distinct anatomical sites.
]]></description>
<dc:creator>Sposito, B.</dc:creator>
<dc:creator>Broggi, A.</dc:creator>
<dc:creator>Pandolfi, L.</dc:creator>
<dc:creator>Crotta, S.</dc:creator>
<dc:creator>Ferrarese, R.</dc:creator>
<dc:creator>Sisti, S.</dc:creator>
<dc:creator>Clementi, N.</dc:creator>
<dc:creator>Ambrosi, A.</dc:creator>
<dc:creator>Liu, E.</dc:creator>
<dc:creator>Frangipane, V.</dc:creator>
<dc:creator>Saracino, L.</dc:creator>
<dc:creator>Marongiu, L.</dc:creator>
<dc:creator>Facchini, F.</dc:creator>
<dc:creator>Bottazzi, A.</dc:creator>
<dc:creator>Fossali, T.</dc:creator>
<dc:creator>Colombo, R.</dc:creator>
<dc:creator>Clementi, M.</dc:creator>
<dc:creator>Tagliabue, E.</dc:creator>
<dc:creator>Pontiroli, A.</dc:creator>
<dc:creator>Meloni, F.</dc:creator>
<dc:creator>Wack, A.</dc:creator>
<dc:creator>Mancini, N.</dc:creator>
<dc:creator>Zanoni, I.</dc:creator>
<dc:date>2021-03-30</dc:date>
<dc:identifier>doi:10.1101/2021.03.30.437173</dc:identifier>
<dc:title><![CDATA[Severity of SARS-CoV-2 infection as a function of the interferon landscape across the respiratory tract of COVID-19 patients.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.01.437886v1?rss=1">
<title>
<![CDATA[
A SIMPLI (Single-cell Identification from MultiPLexed Images) approach for spatially resolved tissue phenotypingat single-cell resolution. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.01.437886v1?rss=1"
</link>
<description><![CDATA[
Multiplexed imaging technologies enable the study of biological tissues at single-cell resolution while preserving spatial information. Currently, high-dimension imaging data analysis is technology-specific and requires multiple tools, restricting analytical scalability and result reproducibility. Here we present SIMPLI (Single-cell Identification from MultiPlexed Images), a novel, flexible and technology-agnostic software that unifies all steps of multiplexed imaging data analysis. After raw image processing, SIMPLI performs a spatially resolved, single-cell analysis of the tissue slide as wells as cell-independent quantifications of marker expression to investigate features undetectable at the cell level. SIMPLI is highly customisable and can run on desktop computers as well as high-performance computing environments, enabling workflow parallelisation for large datasets. SIMPLI produces multiple tabular and graphical outputs at each step of the analysis. Its containerised implementation and minimum configuration requirements make SIMPLI a portable and reproducible solution for multiplexed imaging data analysis.

SIMPLI is available at: https://github.com/ciccalab/SIMPLI.
]]></description>
<dc:creator>Bortolomeazzi, M.</dc:creator>
<dc:creator>Montorsi, L.</dc:creator>
<dc:creator>Temelkovski, D.</dc:creator>
<dc:creator>Keddar, M. R.</dc:creator>
<dc:creator>Acha-Sagredo, A.</dc:creator>
<dc:creator>Pitcher, M. J.</dc:creator>
<dc:creator>Basso, G.</dc:creator>
<dc:creator>Laghi, L.</dc:creator>
<dc:creator>Rodriguez-Justo, M.</dc:creator>
<dc:creator>Spencer, J.</dc:creator>
<dc:creator>Ciccarelli, F. D.</dc:creator>
<dc:date>2021-04-02</dc:date>
<dc:identifier>doi:10.1101/2021.04.01.437886</dc:identifier>
<dc:title><![CDATA[A SIMPLI (Single-cell Identification from MultiPLexed Images) approach for spatially resolved tissue phenotypingat single-cell resolution.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.02.437896v1?rss=1">
<title>
<![CDATA[
Oncogenic RAS activity predicts response to chemotherapy and outcome in lung adenocarcinoma 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.02.437896v1?rss=1"
</link>
<description><![CDATA[
Activating mutations in the driver oncogene KRAS occur in 32% of lung adenocarcinomas, leading to more aggressive disease and resistance to therapy in preclinical studies. However, the association between KRAS mutational status and patient outcome or response to treatment remains unclear, likely due to additional events modulating RAS pathways. To obtain a broader measure of RAS pathway activation beyond KRAS mutation only, we developed RAS84, a transcriptional signature optimised to capture RAS oncogenic activity in lung adenocarcinoma. Using RAS84 to classify lung cell lines, we show that RAS transcriptional activity outperforms KRAS mutation to predict resistance to chemotherapy drugs in vitro. We report that 84% of lung adenocarcinomas show clear transcriptional evidence of RAS oncogenic activation, falling into four groups characterised by coincident mutation of STK11/LKB1, TP53 or CDKN2A. Given that 65% of these RAS pathway active tumours do not have KRAS mutations, we find that the classifications developed when considering only KRAS mutant tumours have significance in a much broader cohort of patients. Critically, patients in the highest RAS activity groups show adverse clinical outcome and reduced response to chemotherapy. The stratification of patients using gene expression patterns linked to oncogenic RAS signalling activity instead of genetic alterations in cancer genes could ultimately help clinical decision making.
]]></description>
<dc:creator>East, P.</dc:creator>
<dc:creator>Kelly, G.</dc:creator>
<dc:creator>Biswas, D.</dc:creator>
<dc:creator>Marani, M.</dc:creator>
<dc:creator>Hancock, D. C.</dc:creator>
<dc:creator>Creasy, T.</dc:creator>
<dc:creator>Sachsenmeier, K.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>de Carne Trecesson, S.</dc:creator>
<dc:creator>Downward, J.</dc:creator>
<dc:date>2021-04-04</dc:date>
<dc:identifier>doi:10.1101/2021.04.02.437896</dc:identifier>
<dc:title><![CDATA[Oncogenic RAS activity predicts response to chemotherapy and outcome in lung adenocarcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.02.438170v1?rss=1">
<title>
<![CDATA[
Common ALS/FTD risk variants in UNC13A exacerbate its cryptic splicing and loss upon TDP-43 mislocalization 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.02.438170v1?rss=1"
</link>
<description><![CDATA[
Variants within the UNC13A gene have long been known to increase risk of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), two related neurodegenerative diseases defined by mislocalization of the RNA-binding protein TDP-43. Here, we show that TDP-43 depletion induces robust inclusion of a cryptic exon (CE) within UNC13A, a critical synaptic gene, resulting in nonsense-mediated decay and protein loss. Strikingly, two common polymorphisms strongly associated with ALS/FTD risk directly alter TDP-43 binding within the CE or downstream intron, increasing CE inclusion in cultured cells and in patient brains. Our findings, which are the first to demonstrate a genetic link specifically between loss of TDP-43 nuclear function and disease, reveal both the mechanism by which UNC13A variants exacerbate the effects of decreased nuclear TDP-43 function, and provide a promising therapeutic target for TDP-43 proteinopathies.

One-Sentence SummaryShared ALS/FTD risk variants increase the sensitivity of a cryptic exon in the synaptic gene UNC13A to TDP-43 depletion.
]]></description>
<dc:creator>Brown, A.-L.</dc:creator>
<dc:creator>Wilkins, O. G.</dc:creator>
<dc:creator>Keuss, M. J.</dc:creator>
<dc:creator>Hill, S. E.</dc:creator>
<dc:creator>Zanovello, M.</dc:creator>
<dc:creator>Lee, W. C.</dc:creator>
<dc:creator>Lee, F. C. Y.</dc:creator>
<dc:creator>Masino, L.</dc:creator>
<dc:creator>Qi, Y. A.</dc:creator>
<dc:creator>Bryce-Smith, S.</dc:creator>
<dc:creator>Bampton, A.</dc:creator>
<dc:creator>Gatt, A.</dc:creator>
<dc:creator>Phatnani, H.</dc:creator>
<dc:creator>NYGC ALS Consortium,</dc:creator>
<dc:creator>Schiavo, G.</dc:creator>
<dc:creator>Fisher, E. M. C.</dc:creator>
<dc:creator>Raj, T.</dc:creator>
<dc:creator>Secrier, M.</dc:creator>
<dc:creator>Lashley, T.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Buratti, E.</dc:creator>
<dc:creator>Humphrey, J.</dc:creator>
<dc:creator>Ward, M. E.</dc:creator>
<dc:creator>Fratta, P.</dc:creator>
<dc:date>2021-04-04</dc:date>
<dc:identifier>doi:10.1101/2021.04.02.438170</dc:identifier>
<dc:title><![CDATA[Common ALS/FTD risk variants in UNC13A exacerbate its cryptic splicing and loss upon TDP-43 mislocalization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.07.438812v1?rss=1">
<title>
<![CDATA[
Identifying SARS-CoV-2 Antiviral Compounds by Screening for Small Molecule Inhibitors of Nsp14/nsp10 Exoribonuclease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.07.438812v1?rss=1"
</link>
<description><![CDATA[
SARS-CoV-2 is a coronavirus that emerged in 2019 and rapidly spread across the world causing a deadly pandemic with tremendous social and economic costs. Healthcare systems worldwide are under great pressure, and there is urgent need for effective antiviral treatments. The only currently approved antiviral treatment for COVID-19 is remdesivir, an inhibitor of viral genome replication. SARS-CoV-2 proliferation relies on the enzymatic activities of the non-structural proteins (nsp), which makes them interesting targets for the development of new antiviral treatments. With the aim to identify novel SARS-CoV-2 antivirals, we have purified the exoribonuclease/methyltransferase (nsp14) and its cofactor (nsp10) and developed biochemical assays compatible with high-throughput approaches to screen for exoribonuclease inhibitors. We have screened a library of over 5000 commercial compounds and identified patulin and aurintricarboxylic acid (ATA) as inhibitors of nsp14 exoribonuclease in vitro. We found that patulin and ATA inhibit replication of SARS-CoV-2 in a VERO E6 cell-culture model. These two new antiviral compounds will be valuable tools for further coronavirus research as well as potentially contributing to new therapeutic opportunities for COVID-19.
]]></description>
<dc:creator>Basier, C.</dc:creator>
<dc:creator>Basu, S.</dc:creator>
<dc:creator>Beale, R.</dc:creator>
<dc:creator>Bertolin, A. P.</dc:creator>
<dc:creator>Canal, B.</dc:creator>
<dc:creator>Curran, J. F.</dc:creator>
<dc:creator>Deegan, T. D.</dc:creator>
<dc:creator>Diffley, J. F.</dc:creator>
<dc:creator>Drury, L. S.</dc:creator>
<dc:creator>Fujisawa, R.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>Labib, K.</dc:creator>
<dc:creator>McClure, A. W.</dc:creator>
<dc:creator>Milligan, J.</dc:creator>
<dc:creator>Roberts, E.</dc:creator>
<dc:creator>Ulferts, R.</dc:creator>
<dc:creator>Weissmann, F.</dc:creator>
<dc:creator>Wu, M.</dc:creator>
<dc:creator>Zeng, J.</dc:creator>
<dc:date>2021-04-08</dc:date>
<dc:identifier>doi:10.1101/2021.04.07.438812</dc:identifier>
<dc:title><![CDATA[Identifying SARS-CoV-2 Antiviral Compounds by Screening for Small Molecule Inhibitors of Nsp14/nsp10 Exoribonuclease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.07.438811v1?rss=1">
<title>
<![CDATA[
Identifying SARS-CoV-2 Antiviral Compounds by Screening for Small Molecule Inhibitors of Nsp15 Endoribonuclease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.07.438811v1?rss=1"
</link>
<description><![CDATA[
SARS-CoV-2 is responsible for COVID-19, a human disease that has caused over 2 million deaths, stretched health systems to near-breaking point and endangered the economies of countries and families around the world. Antiviral treatments to combat COVID-19 are currently lacking. Remdesivir, the only antiviral drug approved for the treatment of COVID-19, can affect disease severity, but better treatments are needed. SARS-CoV-2 encodes 16 non-structural proteins (nsp) that possess different enzymatic activities with important roles in viral genome replication, transcription and host immune evasion. One key aspect of host immune evasion is performed by the uridine-directed endoribonuclease activity of nsp15. Here we describe the expression and purification of nsp15 recombinant protein. We have developed biochemical assays to follow its activity, and we have found evidence for allosteric behaviour. We screened a custom chemical library of over 5000 compounds to identify nsp15 endoribonuclease inhibitors, and we identified and validated NSC95397 as an inhibitor of nsp15 endoribonuclease in vitro. Although NSC95397 did not inhibit SARS-CoV-2 growth in VERO E6 cells, further studies will be required to determine the effect of nsp15 inhibition on host immune evasion.
]]></description>
<dc:creator>Beale, R.</dc:creator>
<dc:creator>Bertolin, A. P.</dc:creator>
<dc:creator>Canal, B.</dc:creator>
<dc:creator>Deegan, T. D.</dc:creator>
<dc:creator>Diffley, J. F.</dc:creator>
<dc:creator>Drury, L. S.</dc:creator>
<dc:creator>Fujisawa, R.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>Labib, K.</dc:creator>
<dc:creator>McClure, A. W.</dc:creator>
<dc:creator>Ulferts, R.</dc:creator>
<dc:creator>Weissmann, F.</dc:creator>
<dc:creator>Wu, M.</dc:creator>
<dc:creator>Zeng, J.</dc:creator>
<dc:date>2021-04-08</dc:date>
<dc:identifier>doi:10.1101/2021.04.07.438811</dc:identifier>
<dc:title><![CDATA[Identifying SARS-CoV-2 Antiviral Compounds by Screening for Small Molecule Inhibitors of Nsp15 Endoribonuclease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.07.438804v1?rss=1">
<title>
<![CDATA[
Identifying SARS-CoV-2 Antiviral Compounds by Screening for Small Molecule Inhibitors of Nsp3 Papain-like Protease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.07.438804v1?rss=1"
</link>
<description><![CDATA[
The COVID-19 pandemic has emerged as the biggest life-threatening disease of this century. Whilst vaccination should provide a long-term solution, this is pitted against the constant threat of mutations in the virus rendering the current vaccines less effective. Consequently, small molecule antiviral agents would be extremely useful to complement the vaccination program. The causative agent of COVID-19 is a novel coronavirus, SARS-CoV-2, which encodes at least nine enzymatic activities that all have drug targeting potential. The papain-like protease (PLpro) contained in the nsp3 protein generates viral non-structural proteins from a polyprotein precursor, and cleaves ubiquitin and ISG protein conjugates. Here we describe the expression and purification of PLpro. We developed a protease assay that was used to screen a custom chemical library from which we identified Dihydrotanshinone I and Ro 08-2750 as compounds that inhibit PLpro in protease and isopeptidase assays and also inhibit viral replication in cell culture-based assays.
]]></description>
<dc:creator>Armstrong, L. A.</dc:creator>
<dc:creator>Beale, R.</dc:creator>
<dc:creator>Bineva-Todd, G.</dc:creator>
<dc:creator>Canal, B.</dc:creator>
<dc:creator>Diffley, J. F.</dc:creator>
<dc:creator>Drury, L. S.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>Kulathu, Y.</dc:creator>
<dc:creator>Labib, K.</dc:creator>
<dc:creator>Lim, C. T.</dc:creator>
<dc:creator>Milligan, J.</dc:creator>
<dc:creator>O'Reilly, N.</dc:creator>
<dc:creator>Ozono, E.</dc:creator>
<dc:creator>Tan, K. W.</dc:creator>
<dc:creator>Ulferts, R.</dc:creator>
<dc:creator>Weissmann, F.</dc:creator>
<dc:creator>Wu, M.</dc:creator>
<dc:creator>Zeisner, T. U.</dc:creator>
<dc:creator>Zeng, J.</dc:creator>
<dc:date>2021-04-08</dc:date>
<dc:identifier>doi:10.1101/2021.04.07.438804</dc:identifier>
<dc:title><![CDATA[Identifying SARS-CoV-2 Antiviral Compounds by Screening for Small Molecule Inhibitors of Nsp3 Papain-like Protease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.07.438808v1?rss=1">
<title>
<![CDATA[
Identifying SARS-CoV-2 Antiviral Compounds by Screening for Small Molecule Inhibitors of Nsp13 Helicase 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.07.438808v1?rss=1"
</link>
<description><![CDATA[
The coronavirus disease 2019 (COVID-19) pandemic, which is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a global public health challenge. While the efficacy of vaccines against emerging and future virus variants remains unclear, there is a need for therapeutics. Repurposing existing drugs represents a promising and potentially rapid opportunity to find novel antivirals against SARS-CoV-2. The virus encodes at least nine enzymatic activities that are potential drug targets. Here we have expressed, purified and developed enzymatic assays for SARS-CoV-2 nsp13 helicase, a viral replication protein that is essential for the coronavirus life cycle. We screened a custom chemical library of over 5000 previously characterised pharmaceuticals for nsp13 inhibitors using a FRET-based high-throughput screening (HTS) approach. From this, we have identified FPA-124 and several suramin-related compounds as novel inhibitors of nsp13 helicase activity in vitro. We describe the efficacy of these drugs using assays we developed to monitor SARS-CoV-2 growth in Vero E6 cells.
]]></description>
<dc:creator>Beale, R.</dc:creator>
<dc:creator>Bertolin, A. P.</dc:creator>
<dc:creator>Borg, A.</dc:creator>
<dc:creator>Canal, B.</dc:creator>
<dc:creator>Diffley, J. F.</dc:creator>
<dc:creator>Drury, L. S.</dc:creator>
<dc:creator>Harvey, R.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>Hussain, S.</dc:creator>
<dc:creator>Kjaer, S.</dc:creator>
<dc:creator>McCauley, J.</dc:creator>
<dc:creator>McCoy, L.</dc:creator>
<dc:creator>Milligan, J.</dc:creator>
<dc:creator>Posse, V.</dc:creator>
<dc:creator>Roustan, C.</dc:creator>
<dc:creator>Ulferts, R.</dc:creator>
<dc:creator>Weissmann, F.</dc:creator>
<dc:creator>Wu, M.</dc:creator>
<dc:creator>Zeng, J.</dc:creator>
<dc:date>2021-04-08</dc:date>
<dc:identifier>doi:10.1101/2021.04.07.438808</dc:identifier>
<dc:title><![CDATA[Identifying SARS-CoV-2 Antiviral Compounds by Screening for Small Molecule Inhibitors of Nsp13 Helicase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.07.438807v1?rss=1">
<title>
<![CDATA[
Identifying SARS-CoV-2 Antiviral Compounds by Screening for Small Molecule Inhibitors of Nsp12/7/8 RNA-dependent RNA Polymerase 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.07.438807v1?rss=1"
</link>
<description><![CDATA[
The coronavirus disease 2019 (COVID-19) global pandemic has turned into the largest public health and economic crisis in recent history impacting virtually all sectors of society. There is a need for effective therapeutics to battle the ongoing pandemic. Repurposing existing drugs with known pharmacological safety profiles is a fast and cost-effective approach to identify novel treatments. The COVID-19 etiologic agent is the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a single-stranded positive-sense RNA virus. Coronaviruses rely on the enzymatic activity of the replication-transcription complex (RTC) to multiply inside host cells. The RTC core catalytic component is the RNA-dependent RNA polymerase (RdRp) holoenzyme. The RdRp is one of the key druggable targets for CoVs due to its essential role in viral replication, high degree of sequence and structural conservation and the lack of homologs in human cells. Here, we have expressed, purified and biochemically characterised active SARS-CoV-2 RdRp complexes. We developed a novel fluorescence resonance energy transfer (FRET)-based strand displacement assay for monitoring SARS-CoV-2 RdRp activity suitable for a high-throughput format. As part of a larger research project to identify inhibitors for all the enzymatic activities encoded by SARS-CoV-2, we used this assay to screen a custom chemical library of over 5000 approved and investigational compounds for novel SARS-CoV-2 RdRp inhibitors. We identified 3 novel compounds (GSK-650394, C646 and BH3I-1) and confirmed suramin and suramin-like compounds as in vitro SARS-CoV-2 RdRp activity inhibitors. We also characterised the antiviral efficacy of these drugs in cell-based assays that we developed to monitor SARS-CoV-2 growth.
]]></description>
<dc:creator>Beale, R.</dc:creator>
<dc:creator>Bertolin, A. P.</dc:creator>
<dc:creator>Canal, B.</dc:creator>
<dc:creator>Diffley, J. F.</dc:creator>
<dc:creator>Drury, L. S.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>Milligan, J.</dc:creator>
<dc:creator>Posse, V.</dc:creator>
<dc:creator>Ulferts, R.</dc:creator>
<dc:creator>Weissmann, F.</dc:creator>
<dc:creator>Wu, M.</dc:creator>
<dc:creator>Zeng, J.</dc:creator>
<dc:date>2021-04-08</dc:date>
<dc:identifier>doi:10.1101/2021.04.07.438807</dc:identifier>
<dc:title><![CDATA[Identifying SARS-CoV-2 Antiviral Compounds by Screening for Small Molecule Inhibitors of Nsp12/7/8 RNA-dependent RNA Polymerase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.07.438806v1?rss=1">
<title>
<![CDATA[
Identifying SARS-CoV-2 Antiviral Compounds by Screening for Small Molecule Inhibitors of nsp5 Main Protease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.07.438806v1?rss=1"
</link>
<description><![CDATA[
The coronavirus 2019 (COVID-19) pandemic, caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), spread around the world with unprecedented health and socio-economic effects for the global population. While different vaccines are now being made available, very few antiviral drugs have been approved. The main viral protease (nsp5) of SARS-CoV-2 provides an excellent target for antivirals, due to its essential and conserved function in the viral replication cycle. We have expressed, purified and developed assays for nsp5 protease activity. We screened the nsp5 protease against a custom chemical library of over 5,000 characterised pharmaceuticals. We identified calpain inhibitor I and three different peptidyl fluoromethylketones (FMK) as inhibitors of nsp5 activity in vitro, with IC50 values in the low micromolar range. By altering the sequence of our peptidomimetic FMK inhibitors to better mimic the substrate sequence of nsp5, we generated an inhibitor with a subnanomolar IC50. Calpain inhibitor I inhibited viral infection in monkey-derived Vero E6 cells, with an EC50 in the low micromolar range. The most potent and commercially available peptidyl-FMK compound inhibited viral growth in Vero E6 cells to some extent, while our custom peptidyl FMK inhibitor offered a marked antiviral improvement.
]]></description>
<dc:creator>Basier, C.</dc:creator>
<dc:creator>Beale, R.</dc:creator>
<dc:creator>Bineva-Todd, G.</dc:creator>
<dc:creator>Canal, B.</dc:creator>
<dc:creator>Curran, J. F.</dc:creator>
<dc:creator>Deegan, T. D.</dc:creator>
<dc:creator>Diffley, J. F.</dc:creator>
<dc:creator>Fujisawa, R.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>Joshi, D.</dc:creator>
<dc:creator>Labib, K.</dc:creator>
<dc:creator>Lim, C. T.</dc:creator>
<dc:creator>Milligan, J.</dc:creator>
<dc:creator>Nagaraj, H.</dc:creator>
<dc:creator>O'Reilly, N.</dc:creator>
<dc:creator>Papageorgiou, G.</dc:creator>
<dc:creator>Soudy, C.</dc:creator>
<dc:creator>Tan, K. W.</dc:creator>
<dc:creator>Ulferts, R.</dc:creator>
<dc:creator>Weissmann, F.</dc:creator>
<dc:creator>Wu, M.</dc:creator>
<dc:creator>Zeisner, T. U.</dc:creator>
<dc:date>2021-04-08</dc:date>
<dc:identifier>doi:10.1101/2021.04.07.438806</dc:identifier>
<dc:title><![CDATA[Identifying SARS-CoV-2 Antiviral Compounds by Screening for Small Molecule Inhibitors of nsp5 Main Protease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.07.438810v1?rss=1">
<title>
<![CDATA[
Identification of SARS-CoV-2 Antiviral Compounds by Screening for Small Molecule Inhibitors of the nsp14 RNA Cap Methyltransferase 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.07.438810v1?rss=1"
</link>
<description><![CDATA[
The COVID-19 pandemic has presented itself as one of the most critical public health challenges of the century, with SARS-CoV-2 being the third member of the Coronaviridae family to cause fatal disease in humans. There is currently only one antiviral compound, remdesivir, that can be used for the treatment of COVID-19. In order to identify additional potential therapeutics, we investigated the enzymatic proteins encoded in the SARS-CoV-2 genome. In this study, we focussed on the viral RNA cap methyltransferases, which play a key role in enabling viral protein translation and facilitating viral escape from the immune system. We expressed and purified both the guanine-N7 methyltransferase nsp14, and the nsp16 2-O-methyltransferase with its activating cofactor, nsp10. We performed an in vitro high-throughput screen for inhibitors of nsp14 using a custom compound library of over 5,000 pharmaceutical compounds that have previously been characterised in either clinical or basic research. We identified 4 compounds as potential inhibitors of nsp14, all of which also show antiviral capacity in a cell based model of SARS-CoV-2 infection. Three of the 4 compounds also exhibited synergistic effects on viral replication with remdesivir.
]]></description>
<dc:creator>Basier, C.</dc:creator>
<dc:creator>Basu, S.</dc:creator>
<dc:creator>Beale, R.</dc:creator>
<dc:creator>Canal, B.</dc:creator>
<dc:creator>Cowling, V. H.</dc:creator>
<dc:creator>Curran, J. F.</dc:creator>
<dc:creator>Deegan, T. D.</dc:creator>
<dc:creator>Diffley, J. F.</dc:creator>
<dc:creator>Drury, L. S.</dc:creator>
<dc:creator>Fujisawa, R.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>Labib, K.</dc:creator>
<dc:creator>Lim, C. T.</dc:creator>
<dc:creator>Mak, T.</dc:creator>
<dc:creator>McClure, A. W.</dc:creator>
<dc:creator>Roberts, E.</dc:creator>
<dc:creator>Tan, K. W.</dc:creator>
<dc:creator>Ulferts, R.</dc:creator>
<dc:creator>Weissmann, F.</dc:creator>
<dc:creator>Wu, M.</dc:creator>
<dc:creator>Zeisner, T. U.</dc:creator>
<dc:date>2021-04-08</dc:date>
<dc:identifier>doi:10.1101/2021.04.07.438810</dc:identifier>
<dc:title><![CDATA[Identification of SARS-CoV-2 Antiviral Compounds by Screening for Small Molecule Inhibitors of the nsp14 RNA Cap Methyltransferase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.08.439054v1?rss=1">
<title>
<![CDATA[
Image-based deep learning reveals the responses of human motor neurons to stress and ALS 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.08.439054v1?rss=1"
</link>
<description><![CDATA[
Although morphological attributes of cells and their substructures are recognized readouts of physiological or pathophysiological states, these have been relatively understudied in amyotrophic lateral sclerosis (ALS) research. In this study we integrate multichannel fluorescence high-content microscopy data with deep-learning imaging methods to reveal - directly from unsegmented images - novel neurite-associated morphological perturbations associated with (ALS-causing) VCP-mutant human motor neurons (MNs). Surprisingly, we reveal that previously unrecognized disease-relevant information is withheld in broadly used and often considered  generic biological markers of nuclei (DAPI) and neurons ({beta}III-tubulin). Additionally, we identify changes within the information content of ALS-related RNA binding protein (RBP) immunofluorescence imaging that is captured in VCP-mutant MN cultures. Furthermore, by analyzing MN cultures exposed to different extrinsic stressors, we show that heat stress recapitulates key aspects of ALS. Our study therefore reveals disease-relevant information contained in a range of both generic and more specific fluorescent markers, and establishes the use of image-based deep learning methods for rapid, automated and unbiased testing of biological hypotheses.
]]></description>
<dc:creator>Verzat, C.</dc:creator>
<dc:creator>Harley, J.</dc:creator>
<dc:creator>Patani, R.</dc:creator>
<dc:creator>Luisier, R.</dc:creator>
<dc:date>2021-04-09</dc:date>
<dc:identifier>doi:10.1101/2021.04.08.439054</dc:identifier>
<dc:title><![CDATA[Image-based deep learning reveals the responses of human motor neurons to stress and ALS]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.09.439171v1?rss=1">
<title>
<![CDATA[
Rad53 checkpoint kinase regulation of DNA replication fork rate via Mrc1 phosphorylation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.09.439171v1?rss=1"
</link>
<description><![CDATA[
The Rad53 DNA checkpoint protein kinase plays multiple roles in the budding yeast cell response to DNA replication stress. Key amongst these is its enigmatic role in safeguarding DNA replication forks. Using DNA replication reactions reconstituted with purified proteins, we show Rad53 phosphorylation of Sld3/7 or Dbf4-dependent kinase blocks replication initiation whilst phosphorylation of Mrc1 or Mcm10 slows elongation. Mrc1 phosphorylation is necessary and sufficient to slow replication forks in complete reactions; Mcm10 phosphorylation can also slow replication forks, but only in the absence of unphosphorylated Mrc1. Mrc1 stimulates the unwinding rate of the replicative helicase, CMG, and Rad53 phosphorylation of Mrc1 prevents this. We show that a phosphorylation-mimicking Mrc1 mutant cannot stimulate replication in vitro and partially rescues the sensitivity of a rad53 null mutant to genotoxic stress in vivo. Our results show that Rad53 protects replication forks in part by antagonising Mrc1 stimulation of CMG unwinding.
]]></description>
<dc:creator>McClure, A. W.</dc:creator>
<dc:creator>Diffley, J.</dc:creator>
<dc:date>2021-04-10</dc:date>
<dc:identifier>doi:10.1101/2021.04.09.439171</dc:identifier>
<dc:title><![CDATA[Rad53 checkpoint kinase regulation of DNA replication fork rate via Mrc1 phosphorylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.09.439198v1?rss=1">
<title>
<![CDATA[
Multiple 9-1-1 complexes promote homolog synapsis, DSB repair, and ATR signaling during mammalian meiosis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.09.439198v1?rss=1"
</link>
<description><![CDATA[
DNA damage response mechanisms have meiotic roles that ensure successful gamete formation. While completion of meiotic double-strand break (DSB) repair requires the canonical RAD9A-RAD1-HUS1 (9A-1-1) complex, mammalian meiocytes also express RAD9A and HUS1 paralogs, RAD9B and HUS1B, predicted to form alternative 9-1-1 complexes. The RAD1 subunit is shared by all predicted 9-1-1 complexes and localizes to meiotic chromosomes even in the absence of HUS1 and RAD9A. Here we report that testis-specific RAD1 disruption resulted in impaired DSB repair, germ cell depletion and infertility. Unlike Hus1 or Rad9a disruption, Rad1 loss also caused defects in homolog synapsis, ATR signaling and meiotic sex chromosome inactivation. Comprehensive testis phosphoproteomics revealed that RAD1 and ATR coordinately regulate numerous proteins involved in DSB repair, meiotic silencing, synaptonemal complex formation, and cohesion. Together, these results establish critical roles for both canonical and alternative 9-1-1 complexes in meiotic ATR activation and successful prophase I completion.
]]></description>
<dc:creator>Pereira, C.</dc:creator>
<dc:creator>Arroyo-Martinez, G. A.</dc:creator>
<dc:creator>Guo, M. Z.</dc:creator>
<dc:creator>Kelly, E. R.</dc:creator>
<dc:creator>Grive, K. J.</dc:creator>
<dc:creator>Mahadevaiah, S. K.</dc:creator>
<dc:creator>Sims, J.</dc:creator>
<dc:creator>Faca, V. M.</dc:creator>
<dc:creator>Tsai, C.</dc:creator>
<dc:creator>Schiltz, C. J.</dc:creator>
<dc:creator>Wit, N.</dc:creator>
<dc:creator>Jacobs, H.</dc:creator>
<dc:creator>Clark, N. L.</dc:creator>
<dc:creator>Freire, R.</dc:creator>
<dc:creator>Turner, J.</dc:creator>
<dc:creator>Lyndaker, A. M.</dc:creator>
<dc:creator>Brieno-Enriquez, M. A.</dc:creator>
<dc:creator>Cohen, P. E.</dc:creator>
<dc:creator>Smolka, M. B.</dc:creator>
<dc:creator>Weiss, R. S.</dc:creator>
<dc:date>2021-04-10</dc:date>
<dc:identifier>doi:10.1101/2021.04.09.439198</dc:identifier>
<dc:title><![CDATA[Multiple 9-1-1 complexes promote homolog synapsis, DSB repair, and ATR signaling during mammalian meiosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.12.439474v1?rss=1">
<title>
<![CDATA[
Single cell transcriptome profiling of the human developing spinal cord reveals a conserved genetic programme with human specific features 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.12.439474v1?rss=1"
</link>
<description><![CDATA[
The spinal cord receives input from peripheral sensory neurons and controls motor output by regulating muscle innervating motor neurons. These functions are carried out by neural circuits comprising molecularly and physiologically distinct neuronal subtypes that are generated in a characteristic spatial-temporal arrangement from progenitors in the embryonic neural tube. The systematic mapping of gene expression in mouse embryos has provided insight into the diversity and complexity of cells in the neural tube. For human embryos, however, less information has been available. To address this, we used single cell mRNA sequencing to profile cervical and thoracic regions in four human embryos of Carnegie Stages (CS) CS12, CS14, CS17 and CS19 from Gestational Weeks (W) 4-7. In total we recovered the transcriptomes of 71,219 cells. Analysis of progenitor and neuronal populations from the neural tube, as well as cells of the peripheral nervous system, in dorsal root ganglia adjacent to the neural tube, identified dozens of distinct cell types and facilitated the reconstruction of the differentiation pathways of specific neuronal subtypes. Comparison with existing mouse datasets revealed the overall similarity of mouse and human neural tube development while highlighting specific features that differed between species. These data provide a catalogue of gene expression and cell type identity in the developing neural tube that will support future studies of sensory and motor control systems and can be explored at https://shiny.crick.ac.uk/scviewer/neuraltube/.
]]></description>
<dc:creator>Rayon, T.</dc:creator>
<dc:creator>Maizels, R. J.</dc:creator>
<dc:creator>Barrington, C.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:date>2021-04-12</dc:date>
<dc:identifier>doi:10.1101/2021.04.12.439474</dc:identifier>
<dc:title><![CDATA[Single cell transcriptome profiling of the human developing spinal cord reveals a conserved genetic programme with human specific features]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.14.437846v1?rss=1">
<title>
<![CDATA[
Single-shot phase contrast microscopy using polarisation-resolved differential phase contrast 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.14.437846v1?rss=1"
</link>
<description><![CDATA[
We present a robust, low-cost single-shot implementation of differential phase microscopy utilising a polarisation-sensitive camera to simultaneously acquire 4 images from which the phase gradients and quantitative phase image can be calculated. This polarisation-resolved differential phase contrast (pDPC) microscopy technique can be interleaved with single-shot imaging polarimetry.
]]></description>
<dc:creator>Kalita, R.</dc:creator>
<dc:creator>Flanagan, W.</dc:creator>
<dc:creator>Lightley, J.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Alexandrov, Y.</dc:creator>
<dc:creator>Garcia, E.</dc:creator>
<dc:creator>Hintze, M.</dc:creator>
<dc:creator>Barkoulas, M.</dc:creator>
<dc:creator>Dunsby, C.</dc:creator>
<dc:creator>French, P. M. W.</dc:creator>
<dc:date>2021-04-14</dc:date>
<dc:identifier>doi:10.1101/2021.04.14.437846</dc:identifier>
<dc:title><![CDATA[Single-shot phase contrast microscopy using polarisation-resolved differential phase contrast]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.15.439906v1?rss=1">
<title>
<![CDATA[
A suite of activity-based probes to dissect the KLK activome in drug-resistant prostate cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.15.439906v1?rss=1"
</link>
<description><![CDATA[
Kallikrein-related peptidases (KLKs) are a family of secreted serine proteases, which form a network - the KLK activome - with an important role in proteolysis and signaling. In prostate cancer (PCa), increased KLK activity promotes tumor growth and metastasis through multiple biochemical pathways, and specific quantification and tracking of changes in the KLK activome could contribute to validation of KLKs as potential drug targets. Herein we report a technology platform based on novel activity-based probes (ABPs) and inhibitors with unprecedented potency and selectivity enabling simultaneous orthogonal analysis of KLK2, KLK3 and KLK14 activity in hormone-responsive PCa cell lines and tumor homogenates. Using selective inhibitors and multiplexed fluorescent activity-based protein profiling (ABPP) we dissect the KLK activome in PCa cells and show that increased KLK14 activity leads to a migratory phenotype. Furthermore, using biotinylated ABPs we show that active KLK molecules are secreted into the bone microenvironment by PCa cells following stimulation by osteoblasts suggesting KLK-mediated signaling mechanisms could contribute to PCa metastasis to bone. Together our findings show that ABPP is a powerful approach to dissect dysregulation of the KLK activome as a promising and previously underappreciated therapeutic target in advanced PCa.
]]></description>
<dc:creator>Lovell, S.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Kryza, T.</dc:creator>
<dc:creator>Neodo, A.</dc:creator>
<dc:creator>Bock, N.</dc:creator>
<dc:creator>Williams, E. D.</dc:creator>
<dc:creator>Engelsberger, E.</dc:creator>
<dc:creator>Xu, C.</dc:creator>
<dc:creator>Bakker, A. T.</dc:creator>
<dc:creator>De Vita, E.</dc:creator>
<dc:creator>Maneiro, M.</dc:creator>
<dc:creator>Tanaka, R. J.</dc:creator>
<dc:creator>Bevan, C. L.</dc:creator>
<dc:creator>Clements, J. A.</dc:creator>
<dc:creator>Tate, E. W.</dc:creator>
<dc:date>2021-04-15</dc:date>
<dc:identifier>doi:10.1101/2021.04.15.439906</dc:identifier>
<dc:title><![CDATA[A suite of activity-based probes to dissect the KLK activome in drug-resistant prostate cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.16.440164v1?rss=1">
<title>
<![CDATA[
A gene regulatory network for neural induction 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.16.440164v1?rss=1"
</link>
<description><![CDATA[
During early vertebrate development, signals from a special region of the embryo, the organizer, can re-direct the fate of non-neural ectoderm cells to form a complete, patterned nervous system. This is called neural induction and has generally been imagined as a single signalling event, causing a switch of fate. Here we undertake a comprehensive analysis, in very fine time-course, of the events following exposure of ectoderm to the organizer. Using transcriptomics and epigenomics we generate a Gene Regulatory Network comprising 175 transcriptional regulators and 5,614 predicted interactions between them, with fine temporal dynamics from initial exposure to the signals to expression of mature neural plate markers. Using in situ hybridization, single-cell RNA-sequencing and reporter assays we show that neural induction by a grafted organizer closely resembles normal neural plate development. The study is accompanied by a comprehensive resource including information about conservation of the predicted enhancers in different vertebrate systems.
]]></description>
<dc:creator>Trevers, K. E.</dc:creator>
<dc:creator>Lu, H.-C.</dc:creator>
<dc:creator>Yang, Y.</dc:creator>
<dc:creator>Thiery, A.</dc:creator>
<dc:creator>Strobl, A. C.</dc:creator>
<dc:creator>Palinkasova, B.</dc:creator>
<dc:creator>de Oliveira, N. M. M.</dc:creator>
<dc:creator>de Almeida, I. M.</dc:creator>
<dc:creator>Khan, M. A. F.</dc:creator>
<dc:creator>Moncaut, N.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:creator>Dale, L.</dc:creator>
<dc:creator>Streit, A.</dc:creator>
<dc:creator>Stern, C. D.</dc:creator>
<dc:date>2021-04-16</dc:date>
<dc:identifier>doi:10.1101/2021.04.16.440164</dc:identifier>
<dc:title><![CDATA[A gene regulatory network for neural induction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.16.440179v1?rss=1">
<title>
<![CDATA[
Developmental regulation of neuronal gene expression by Elongator complex protein 1 dosage. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.16.440179v1?rss=1"
</link>
<description><![CDATA[
Elongator is a highly conserved protein complex required for transcriptional elongation, intracellular transport and translation. Elongator complex protein 1 (ELP1) is the scaffolding protein of Elongator and is essential for its assembly and stability. Familial dysautonomia (FD), a hereditary sensory and autonomic neuropathy, is caused by a mutation in ELP1 that lead to a tissue-specific reduction of ELP1 protein. Our work to generate a phenotypic mouse model for FD led to the discovery that homozygous deletion of the mouse Elp1 gene leads to embryonic lethality prior to mid-gestation. Given that FD is caused by a reduction, not loss, of ELP1, we generated two new mouse models by introducing different copy numbers of the human FD ELP1 transgene into the Elp1 knockout mouse (Elp1-/-) and observed that human ELP1 expression rescues embryonic development in a dose dependent manner. We then conducted a comprehensive transcriptome analysis in mouse embryos to identify genes and pathways whose expression correlates with the amount of ELP1. We found that ELP1 is essential for the expression of genes responsible for the formation and development of the nervous system. Further, gene length analysis of the differentially expressed genes showed that the loss of Elp1 mainly impacts the expression of long genes and that by gradually restoring Elongator their expression is progressively rescued. Finally, through evaluation of co-expression modules, we identified gene sets with unique expression patterns that depended on ELP1 expression. Overall, this study highlights the crucial role of ELP1 during early embryonic neuronal development and reveals gene networks and biological pathways that are regulated by Elongator.
]]></description>
<dc:creator>Morini, E.</dc:creator>
<dc:creator>Gao, D.</dc:creator>
<dc:creator>Logan, E. M.</dc:creator>
<dc:creator>Salani, M.</dc:creator>
<dc:creator>Krauson, A. J.</dc:creator>
<dc:creator>Chekuri, A.</dc:creator>
<dc:creator>Chen, Y.-T.</dc:creator>
<dc:creator>Ragavendran, A.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Erdin, S.</dc:creator>
<dc:creator>Stortchevoi, A.</dc:creator>
<dc:creator>Svejstrup, J. Q.</dc:creator>
<dc:creator>Talkowski, M. E.</dc:creator>
<dc:creator>Slaugenhaupt, S. A.</dc:creator>
<dc:date>2021-04-16</dc:date>
<dc:identifier>doi:10.1101/2021.04.16.440179</dc:identifier>
<dc:title><![CDATA[Developmental regulation of neuronal gene expression by Elongator complex protein 1 dosage.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.19.440412v1?rss=1">
<title>
<![CDATA[
Aberrant integration of Hepatitis B virus DNA promotes major restructuring of human hepatocellular carcinoma genome architecture 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.19.440412v1?rss=1"
</link>
<description><![CDATA[
Most cancers are characterized by the somatic acquisition of genomic rearrangements during tumour evolution that eventually drive the oncogenesis. There are different mutational mechanisms causing structural variation, some of which are specific to particular cancer types. Here, using multiplatform sequencing technologies, we identify and characterize a remarkable mutational mechanism in human hepatocellular carcinoma caused by Hepatitis B virus, by which DNA molecules from the virus are inserted into the tumour genome causing dramatic changes in its configuration, including non-homologous chromosomal fusions and megabase-size telomeric deletions. This aberrant mutational process, present in at least 8% of all HCC tumours, is active early during liver cancer evolution and can provide the driver rearrangements that a cancer clone requires to survive and grow.
]]></description>
<dc:creator>Alvarez, E. G.</dc:creator>
<dc:creator>Demeulemeester, J.</dc:creator>
<dc:creator>Jolly, C.</dc:creator>
<dc:creator>Garcia-Souto, D.</dc:creator>
<dc:creator>Otero, P.</dc:creator>
<dc:creator>Pequeno, A.</dc:creator>
<dc:creator>Zamora, J.</dc:creator>
<dc:creator>Tojo, M.</dc:creator>
<dc:creator>Temes, J.</dc:creator>
<dc:creator>Baez-Ortega, A.</dc:creator>
<dc:creator>Rodriguez-Martin, B.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Oitaben, A.</dc:creator>
<dc:creator>Bruzos, A. L.</dc:creator>
<dc:creator>Martinez-Fernandez, M.</dc:creator>
<dc:creator>Haase, K.</dc:creator>
<dc:creator>Santamarina, M.</dc:creator>
<dc:creator>Zumalave, S.</dc:creator>
<dc:creator>Abal, R.</dc:creator>
<dc:creator>Rodriguez-Castro, J.</dc:creator>
<dc:creator>Rodriguez-Casanova, A.</dc:creator>
<dc:creator>Diaz-Lagares, A.</dc:creator>
<dc:creator>Raine, K.</dc:creator>
<dc:creator>Butler, A. P.</dc:creator>
<dc:creator>Ono, A.</dc:creator>
<dc:creator>Aikata, H.</dc:creator>
<dc:creator>Chayama, K.</dc:creator>
<dc:creator>Ueno, M.</dc:creator>
<dc:creator>Hayami, S.</dc:creator>
<dc:creator>Yamaue, H.</dc:creator>
<dc:creator>Blanco, M. G.</dc:creator>
<dc:creator>Forns, X.</dc:creator>
<dc:creator>Rivas, C.</dc:creator>
<dc:creator>Perez-del-Pulgar, S.</dc:creator>
<dc:creator>Torres-Ruiz, R.</dc:creator>
<dc:creator>Rodriguez Perales, S.</dc:creator>
<dc:creator>Garaigorta, U.</dc:creator>
<dc:creator>Nakagawa, H.</dc:creator>
<dc:creator>Campbell, P. J.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>Tubio, J.</dc:creator>
<dc:date>2021-04-19</dc:date>
<dc:identifier>doi:10.1101/2021.04.19.440412</dc:identifier>
<dc:title><![CDATA[Aberrant integration of Hepatitis B virus DNA promotes major restructuring of human hepatocellular carcinoma genome architecture]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.20.440373v1?rss=1">
<title>
<![CDATA[
Early immune pressure imposed by tissue resident memory T cells sculpts tumour evolution in non-small cell lung cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.20.440373v1?rss=1"
</link>
<description><![CDATA[
Tissue-resident memory T cells (TRM) provide immune defence against local infection and can inhibit cancer progression. However, it is unclear to what extent chronic inflammation impacts TRM activation and how the immune pressure exerted by TRM affects developing tumours in humans. We performed deep profiling of lung cancers arising in never-smokers (NS) and ever-smokers (ES), finding evidence of enhanced TRM immunosurveillance in ES lung. Only tumours arising in ES patients underwent clonal immune escape, even when evaluating cancers with similar tumour mutational burden to NS patients, suggesting that the timing of immune pressure exerted by TRM is a critical factor in the evolution of tumour immune evasion. Tumours grown in T cell quiescent NS lungs displayed little evidence of immune evasion and had fewer neoantigens with low diversity, paradoxically making them amenable to treatment with agonist of the costimulatory molecule, ICOS. These data demonstrate local environmental insults enhance TRM immunosurveillance of human tissue, shape the evolution of tumour immunogenicity and that this interplay informs effective immunotherapeutic modalities.
]]></description>
<dc:creator>Weeden, C. E.</dc:creator>
<dc:creator>Gayevskiy, V.</dc:creator>
<dc:creator>Trussart, M.</dc:creator>
<dc:creator>Marceaux, C.</dc:creator>
<dc:creator>Tubau Ribera, N.</dc:creator>
<dc:creator>Batey, D.</dc:creator>
<dc:creator>Teh, C. E.</dc:creator>
<dc:creator>Mitchell, A. J.</dc:creator>
<dc:creator>Antippa, P.</dc:creator>
<dc:creator>Leong, T.</dc:creator>
<dc:creator>Steinfort, D.</dc:creator>
<dc:creator>Irving, L.</dc:creator>
<dc:creator>Gordon, C. L.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Speed, T. P.</dc:creator>
<dc:creator>Gray, D. H.</dc:creator>
<dc:creator>Asselin-Labat, M.-L.</dc:creator>
<dc:date>2021-04-21</dc:date>
<dc:identifier>doi:10.1101/2021.04.20.440373</dc:identifier>
<dc:title><![CDATA[Early immune pressure imposed by tissue resident memory T cells sculpts tumour evolution in non-small cell lung cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.21.440736v1?rss=1">
<title>
<![CDATA[
A systematic benchmark of Nanopore long read RNA sequencing for transcript level analysis in human cell lines 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.21.440736v1?rss=1"
</link>
<description><![CDATA[
The human genome contains more than 200,000 gene isoforms. However, different isoforms can be highly similar, and with an average length of 1.5kb remain difficult to study with short read sequencing. To systematically evaluate the ability to study the transcriptome at a resolution of individual isoforms we profiled 5 human cell lines with short read cDNA sequencing and Nanopore long read direct RNA, amplification-free direct cDNA, PCR-cDNA sequencing. The long read protocols showed a high level of consistency, with amplification-free RNA and cDNA sequencing being most similar. While short and long reads generated comparable gene expression estimates, they differed substantially for individual isoforms. We find that increased read length improves read-to-transcript assignment, identifies interactions between alternative promoters and splicing, enables the discovery of novel transcripts from repetitive regions, facilitates the quantification of full-length fusion isoforms and enables the simultaneous profiling of m6A RNA modifications when RNA is sequenced directly. Our study demonstrates the advantage of long read RNA sequencing and provides a comprehensive resource that will enable the development and benchmarking of computational methods for profiling complex transcriptional events at isoform-level resolution.
]]></description>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Davidson, N.</dc:creator>
<dc:creator>Wan, Y. K.</dc:creator>
<dc:creator>Patel, H.</dc:creator>
<dc:creator>Yao, F.</dc:creator>
<dc:creator>Low, H. M.</dc:creator>
<dc:creator>Hendra, C.</dc:creator>
<dc:creator>Watten, L.</dc:creator>
<dc:creator>Sim, A.</dc:creator>
<dc:creator>Sawyer, C.</dc:creator>
<dc:creator>Iakovleva, V.</dc:creator>
<dc:creator>Lee, P. L.</dc:creator>
<dc:creator>Xin, L.</dc:creator>
<dc:creator>Ng, H. E. V.</dc:creator>
<dc:creator>Loo, J. M.</dc:creator>
<dc:creator>Ong, X.</dc:creator>
<dc:creator>Ng, H. Q. A.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Koh, W. Q. C.</dc:creator>
<dc:creator>Poon, S. Y. P.</dc:creator>
<dc:creator>Stanojevic, D.</dc:creator>
<dc:creator>Tran, H.-D.</dc:creator>
<dc:creator>Lim, K. H. E.</dc:creator>
<dc:creator>Toh, S. Y.</dc:creator>
<dc:creator>Ewels, P.</dc:creator>
<dc:creator>Ng, H.-H.</dc:creator>
<dc:creator>Iyer, N. G.</dc:creator>
<dc:creator>Thiery, A.</dc:creator>
<dc:creator>Chng, W. J.</dc:creator>
<dc:creator>Chen, L.</dc:creator>
<dc:creator>DasGupta, R.</dc:creator>
<dc:creator>Sikic, M.</dc:creator>
<dc:creator>Chan, Y.-S.</dc:creator>
<dc:creator>Tan, B. O. P.</dc:creator>
<dc:creator>Wan, Y.</dc:creator>
<dc:creator>Tam, W. L.</dc:creator>
<dc:creator>Yu, Q.</dc:creator>
<dc:creator>Khor, C. C.</dc:creator>
<dc:creator>Wuestefeld, T.</dc:creator>
<dc:creator>Pratanwanich, P. N.</dc:creator>
<dc:creator>Love, M. I.</dc:creator>
<dc:creator>Goh, W. S. S.</dc:creator>
<dc:creator>Ng, S.</dc:creator>
<dc:creator>Oshlack, A.</dc:creator>
<dc:creator>Goeke, J.</dc:creator>
<dc:date>2021-04-22</dc:date>
<dc:identifier>doi:10.1101/2021.04.21.440736</dc:identifier>
<dc:title><![CDATA[A systematic benchmark of Nanopore long read RNA sequencing for transcript level analysis in human cell lines]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.23.440888v1?rss=1">
<title>
<![CDATA[
Molecular reconstruction of recurrent evolutionary switching in olfactory receptor specificity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.23.440888v1?rss=1"
</link>
<description><![CDATA[
Olfactory receptor repertoires exhibit remarkable functional diversity, but how these proteins have evolved is poorly understood. Through analysis of extant and ancestrally-reconstructed drosophilid olfactory receptors from the Ionotropic Receptor (IR) family, we investigated evolution of two organic acid-sensing receptors, IR75a and IR75b. Despite their low amino acid identity, we identify a common "hotspot" in their ligand-binding pocket that has a major effect on changing the specificity of both IRs, as well as at least two distinct functional transitions in IR75a during evolution. Ligand-docking into IR models predicts that the hotspot does not contact odor molecules, suggesting that this residue indirectly influences ligand/receptor interactions. Moreover, we show that odor specificity is refined by changes in additional, receptor-specific sites, including those outside the ligand-binding pocket. Our work reveals how a core, common determinant of ligand-tuning acts within epistatic and allosteric networks of substitutions to lead to functional evolution of olfactory receptors.
]]></description>
<dc:creator>Prieto-Godino, L. L.</dc:creator>
<dc:creator>Schmidt, H. R.</dc:creator>
<dc:creator>Benton, R.</dc:creator>
<dc:date>2021-04-23</dc:date>
<dc:identifier>doi:10.1101/2021.04.23.440888</dc:identifier>
<dc:title><![CDATA[Molecular reconstruction of recurrent evolutionary switching in olfactory receptor specificity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.20.440617v1?rss=1">
<title>
<![CDATA[
Sox8 is sufficient to reprogram ectoderm into ear vesicles and associated neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.20.440617v1?rss=1"
</link>
<description><![CDATA[
The vertebrate inner ear arises from a pool of progenitors with the potential to contribute to all the sense organs and cranial ganglia in the head. Here we explore the molecular mechanisms that control ear specification from these precursors. Using a multi-omics approach combined with loss-of-function experiments we identify a core transcriptional circuit that imparts ear identity, along with the first genome-wide characterization of non-coding elements that integrate this information. This analysis places the transcription factor Sox8 at the top of the ear determination network. Introducing Sox8 into cranial ectoderm not only converts non-ear cells into ear progenitors, but also activates the cellular programs for ear morphogenesis and neurogenesis. Thus, Sox8 has the unique ability to remodel transcriptional networks in the cranial ectoderm towards ear identity.
]]></description>
<dc:creator>Buzzi, A. L.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Thiery, A.</dc:creator>
<dc:creator>Delile, J.</dc:creator>
<dc:creator>Streit, A.</dc:creator>
<dc:date>2021-04-20</dc:date>
<dc:identifier>doi:10.1101/2021.04.20.440617</dc:identifier>
<dc:title><![CDATA[Sox8 is sufficient to reprogram ectoderm into ear vesicles and associated neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.30.442156v1?rss=1">
<title>
<![CDATA[
Volumetric Semantic Instance Segmentation of the Plasma Membrane of HeLa Cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.30.442156v1?rss=1"
</link>
<description><![CDATA[
In this work, the unsupervised volumetric semantic segmentation of the plasma membrane of HeLa cells as observed with Serial Block Face Scanning Electron Microscopy is described. The resin background of the images was segmented at different slices of a 3D stack of 518 slices with 8, 192 x 8, 192 pixels each. The background was used to create a distance map which helped identify and rank the cells by their size at each slice. The centroids of the cells detected at different slices were linked to identify them as a single cell that spanned a number of slices. A subset of these cells, i.e., largest ones and those not close to the edges were selected for further processing. The selected cells were then automatically cropped to smaller regions of interest of 2, 000 x 2, 000 x 300 voxels that were treated as cell instances. Then, for each of these volumes the nucleus was segmented and the cell was separated from any neighbouring cells through a series of traditional image processing steps that followed the plasma membrane. The segmentation process was repeated for all the regions selected. For one cell for which the ground truth was available, the algorithm provided excellent results in Accuracy (AC) and Jaccard Index (JI): Nucleus: JI = 0.9665, AC= 0.9975, Cell and Nucleus JI = 0.8711, AC = 0.9655, Cell only JI = 0.8094, AC = 0.9629. A limitation of the algorithm for the plasma membrane segmentation was the presence of background, as in cases of tightly packed cells. When tested for these conditions, the segmentation of the nuclear envelope was still possible. All the code and data are released openly through GitHub, Zenodo and EMPIAR.
]]></description>
<dc:creator>Karabag, C.</dc:creator>
<dc:creator>Jones, M. L.</dc:creator>
<dc:creator>Reyes-Aldasoro, C. C.</dc:creator>
<dc:date>2021-04-30</dc:date>
<dc:identifier>doi:10.1101/2021.04.30.442156</dc:identifier>
<dc:title><![CDATA[Volumetric Semantic Instance Segmentation of the Plasma Membrane of HeLa Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.30.441940v1?rss=1">
<title>
<![CDATA[
Signatures of copy number alterations in human cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.30.441940v1?rss=1"
</link>
<description><![CDATA[
The gains and losses of DNA that emerge as a consequence of mitotic errors and chromosomal instability are prevalent in cancer. These copy number alterations contribute to cancer initiaition, progression and therapeutic resistance. Here, we present a conceptual framework for examining the patterns of copy number alterations in human cancer using whole-genome sequencing, whole-exome sequencing, and SNP6 microarray data making it widely applicable to diverse datasets. Deploying this framework to 9,873 cancers representing 33 human cancer types from the TCGA project revealed a set of 19 copy number signatures that explain the copy number patterns of 93% of TCGA samples. 15 copy number signatures were attributed to biological processes of whole-genome doubling, aneuploidy, loss of heterozygosity, homologous recombination deficiency, and chromothripsis. The aetiology of four copy number signatures are unexplained and some cancer types have unique patterns of amplicon signatures associated with extrachromosomal DNA, disease-specific survival, and gains of proto-oncogenes such as MDM2. In contrast to base-scale mutational signatures, no copy number signature associated with known cancer risk factors. The results provide a foundation for exploring patterns of copy number changes in cancer genomes and synthesise the global landscape of copy number alterations in human cancer by revealing a diversity of mutational processes giving rise to copy number changes.
]]></description>
<dc:creator>Steele, C. D.</dc:creator>
<dc:creator>Abbasi, A.</dc:creator>
<dc:creator>Islam, A. S. M.</dc:creator>
<dc:creator>Khandekar, A.</dc:creator>
<dc:creator>Haase, K.</dc:creator>
<dc:creator>Hames, S.</dc:creator>
<dc:creator>Tarabichi, M.</dc:creator>
<dc:creator>Lesluyes, T.</dc:creator>
<dc:creator>Flanagan, A. M.</dc:creator>
<dc:creator>Mertens, F.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>Alexandrov, L. B.</dc:creator>
<dc:creator>Pillay, N.</dc:creator>
<dc:date>2021-04-30</dc:date>
<dc:identifier>doi:10.1101/2021.04.30.441940</dc:identifier>
<dc:title><![CDATA[Signatures of copy number alterations in human cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.05.01.442251v1?rss=1">
<title>
<![CDATA[
Flow cytometry method for absolute counting and single-cell phenotyping of mycobacteria 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.05.01.442251v1?rss=1"
</link>
<description><![CDATA[
Detection and accurate quantitation of viable Mycobacterium tuberculosis is fundamental to understanding mycobacterial pathogenicity, tuberculosis (TB) disease progression and outcomes; TB transmission; drug action, efficacy and drug resistance. Despite this importance, methods for determining numbers of viable bacilli are limited in accuracy and precision owing to inherent characteristics of mycobacterial cell biology - including the tendency to clump, and "differential" culturability - and technical challenges consequent on handling an infectious pathogen under biosafe conditions. We developed an absolute counting method for mycobacteria in liquid cultures using a bench-top flow cytometer, and the low-cost fluorescent dyes Calcein-AM (CA) and SYBR-gold (SG). During exponential growth CA+ cell counts are highly correlated with CFU counts and can be used as a real-time alternative to simplify the accurate standardisation of inocula for experiments. In contrast to CFU counting, this method can detect and enumerate cell aggregates in samples, which we show are a potential source of variance and bias when using established methods. We show that CFUs comprise a sub-population of intact, metabolically active mycobacterial cells in liquid cultures, with CFU-proportion varying by growth conditions. A pharmacodynamic application of the flow cytometry method, exploring kinetics of fluorescent probe defined subpopulations compared to CFU is demonstrated. Flow cytometry derived Mycobacterium bovis BCG time-kill curves differ for rifampicin and kanamycin versus isoniazid and ethambutol, as do the relative dynamics of discrete morphologically-distinct subpopulations of bacilli revealed by this high-throughput single-cell technique.
]]></description>
<dc:creator>Barr, D. A.</dc:creator>
<dc:creator>Omollo, C. O.</dc:creator>
<dc:creator>Mason, M.</dc:creator>
<dc:creator>Koch, A.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Lalloo, D. G.</dc:creator>
<dc:creator>Meintjes, G.</dc:creator>
<dc:creator>Mizrahi, V.</dc:creator>
<dc:creator>Warner, D. F.</dc:creator>
<dc:creator>Davies, G. R.</dc:creator>
<dc:date>2021-05-01</dc:date>
<dc:identifier>doi:10.1101/2021.05.01.442251</dc:identifier>
<dc:title><![CDATA[Flow cytometry method for absolute counting and single-cell phenotyping of mycobacteria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.05.07.443085v1?rss=1">
<title>
<![CDATA[
Integrated Genome and Transcriptome Analyses Reveal the Mechanism of Genome Instability in Ataxia with Oculomotor Apraxia 2 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.05.07.443085v1?rss=1"
</link>
<description><![CDATA[
Mutations in the SETX gene, which encodes Senataxin, are associated with the progressive neurodegenerative diseases Ataxia with Oculomotor Apraxia 2 (AOA2) and Amyotrophic Lateral Sclerosis 4 (ALS4). To identify the causal defect in AOA2, patient-derived cells and SETX knockouts (human and mouse) were analyzed using integrated genomic and transcriptomic approaches. We observed a genome-wide increase in chromosome instability (gains and losses) within genes and at chromosome fragile sites, resulting in changes to gene expression profiles. Senataxin loss caused increased transcription stress near promoters that correlated with high GCskew and R-loop accumulation at promoter-proximal regions. Notably, the chromosomal regions with gains and losses overlapped with regions of elevated transcription stress. In the absence of Senataxin, we found that Cockayne Syndrome protein CSB was required for the recruitment of the transcription-coupled repair endonucleases (XPG and XPF) and recombination protein RAD52 to target and resolve transcription bubbles containing R-loops, leading to error prone repair and genomic instability. These results show that transcription stress is an important contributor to SETX mutation-associated chromosome fragility and AOA2.
]]></description>
<dc:creator>Kanagaraj, R.</dc:creator>
<dc:creator>Mitter, R.</dc:creator>
<dc:creator>Kantidakis, T.</dc:creator>
<dc:creator>Edwards, M.</dc:creator>
<dc:creator>Benitez, A.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Fu, B.</dc:creator>
<dc:creator>Becherel, O.</dc:creator>
<dc:creator>Yang, F.</dc:creator>
<dc:creator>Lavin, M.</dc:creator>
<dc:creator>Koren, A.</dc:creator>
<dc:creator>Stewart, A.</dc:creator>
<dc:creator>West, S.</dc:creator>
<dc:date>2021-05-08</dc:date>
<dc:identifier>doi:10.1101/2021.05.07.443085</dc:identifier>
<dc:title><![CDATA[Integrated Genome and Transcriptome Analyses Reveal the Mechanism of Genome Instability in Ataxia with Oculomotor Apraxia 2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.05.11.443558v1?rss=1">
<title>
<![CDATA[
Elongation factor ELOF1 drives transcription-coupled repair and prevents genome instability 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.05.11.443558v1?rss=1"
</link>
<description><![CDATA[
Correct transcription is crucial for life. However, DNA damage severely impedes elongating RNA Polymerase II (Pol II), causing transcription inhibition and transcription-replication conflicts. Cells are equipped with intricate mechanisms to counteract the severe consequence of these transcription-blocking lesions (TBLs). However, the exact mechanism and factors involved remain largely unknown. Here, using a genome-wide CRISPR/cas9 screen, we identified elongation factor ELOF1 as an important new factor in the transcription stress response upon DNA damage. We show that ELOF1 has an evolutionary conserved role in Transcription-Coupled Nucleotide Excision Repair (TC-NER), where it promotes recruitment of the TC-NER factors UVSSA and TFIIH to efficiently repair TBLs and resume transcription. Additionally, ELOF1 modulates transcription to protect cells from transcription-mediated replication stress, thereby preserving genome stability. Thus, ELOF1 protects the transcription machinery from DNA damage by two distinct mechanisms.
]]></description>
<dc:creator>Geijer, M.</dc:creator>
<dc:creator>Zhou, D.</dc:creator>
<dc:creator>Selvam, K.</dc:creator>
<dc:creator>Steurer, B.</dc:creator>
<dc:creator>Evers, B.</dc:creator>
<dc:creator>Mukherjee, C.</dc:creator>
<dc:creator>Cugusi, S.</dc:creator>
<dc:creator>van Toorn, M.</dc:creator>
<dc:creator>van der Woude, M.</dc:creator>
<dc:creator>Gong, W.</dc:creator>
<dc:creator>Janssens, R.</dc:creator>
<dc:creator>Raams, A.</dc:creator>
<dc:creator>Lebbink, J.</dc:creator>
<dc:creator>Geverts, B.</dc:creator>
<dc:creator>Plummer, D.</dc:creator>
<dc:creator>Bezstarosti, K.</dc:creator>
<dc:creator>Theil, A.</dc:creator>
<dc:creator>Mitter, R.</dc:creator>
<dc:creator>Houtsmuller, A.</dc:creator>
<dc:creator>Vermeulen, W.</dc:creator>
<dc:creator>Demmers, J.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Lans, H.</dc:creator>
<dc:creator>Bernards, R.</dc:creator>
<dc:creator>Svejstrup, J.</dc:creator>
<dc:creator>Ray Chaudhuri, A.</dc:creator>
<dc:creator>Wyrick, J.</dc:creator>
<dc:creator>Marteijn, J.</dc:creator>
<dc:date>2021-05-11</dc:date>
<dc:identifier>doi:10.1101/2021.05.11.443558</dc:identifier>
<dc:title><![CDATA[Elongation factor ELOF1 drives transcription-coupled repair and prevents genome instability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.05.11.443575v1?rss=1">
<title>
<![CDATA[
The Aspartyl Protease Ddi1 Is Essential for Erythrocyte Invasion by the Malaria Parasite 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.05.11.443575v1?rss=1"
</link>
<description><![CDATA[
Malaria pathology is caused by the exponential replication of Plasmodium parasites in the blood stream. The bottleneck of the parasite life cycle is the invasion of erythrocytes immediately after parasites egress from infected red blood cells. DNA damage-inducible protein 1 (Ddi1) is a conserved eukaryotic proteasome shuttle protein containing an internal retroviral-like protease domain. Using conditional genetics, we now show that the proteolytic activity of the P. falciparum homologue, PfDdi1, is critically required for invasion of red blood cells. Furthermore, PfDdi1 disruption results in the accumulation of highly polyubiquitinated proteins that can be processed by purified PfDdi1 or distant eukaryotic homologues. We also show that PfDdi1 interacts with multiple components of the ubiquitin-proteasome system and that parasites lacking PfDdi1 are more sensitive to proteasome inhibition. Overall, this study establishes PfDdi1 as a key component of the eukaryotic ubiquitin-proteasome system and as a promising antimalarial target.
]]></description>
<dc:creator>Ridewood, S.</dc:creator>
<dc:creator>Dirac-Svejstrup, B.</dc:creator>
<dc:creator>Howell, S. A.</dc:creator>
<dc:creator>Weston, A.</dc:creator>
<dc:creator>Lehmann, C.</dc:creator>
<dc:creator>Patel, A. P.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:creator>Bingham, R.</dc:creator>
<dc:creator>Powell, D.</dc:creator>
<dc:creator>Snijder, A.</dc:creator>
<dc:creator>Svejstrup, J. Q.</dc:creator>
<dc:creator>Deu, E.</dc:creator>
<dc:date>2021-05-11</dc:date>
<dc:identifier>doi:10.1101/2021.05.11.443575</dc:identifier>
<dc:title><![CDATA[The Aspartyl Protease Ddi1 Is Essential for Erythrocyte Invasion by the Malaria Parasite]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.05.12.443934v1?rss=1">
<title>
<![CDATA[
SMCHD1 has separable roles in chromatin architecture and gene silencing that could be targeted in disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.05.12.443934v1?rss=1"
</link>
<description><![CDATA[
The interplay between 3D chromatin architecture and gene silencing is incompletely understood. Here, we report a novel point mutation in the non-canonical SMC protein SMCHD1 that enhances its silencing capacity at endogenous developmental targets and at the facioscapulohumeral muscular dystrophy associated macro-array, D4Z4. Heightened SMCHD1 silencing perturbs developmental Hox gene activation, causing a homeotic transformation in mice. Paradoxically, the mutant SMCHD1 appears to enhance insulation against another epigenetic regulator complex, PRC2, while depleting long range chromatin interactions akin to what is observed in the absence of SMCHD1. These data suggest that SMCHD1s role in long range chromatin interactions is not directly linked to gene silencing or insulating the chromatin, refining the model for how the different levels of SMCHD1-mediated chromatin regulation interact to bring about gene silencing in normal development and disease.
]]></description>
<dc:creator>Tapia del Fierro, A.</dc:creator>
<dc:creator>den Hamer, B.</dc:creator>
<dc:creator>Jansz, N.</dc:creator>
<dc:creator>Chen, K.</dc:creator>
<dc:creator>Beck, T.</dc:creator>
<dc:creator>Vanyai, H.</dc:creator>
<dc:creator>Benetti, N.</dc:creator>
<dc:creator>Gurzau, A. D.</dc:creator>
<dc:creator>Daxinger, L.</dc:creator>
<dc:creator>Xue, S.</dc:creator>
<dc:creator>Nguyen Ly, T. T.</dc:creator>
<dc:creator>Wanigasuriya, I.</dc:creator>
<dc:creator>Breslin, K.</dc:creator>
<dc:creator>Oey, H.</dc:creator>
<dc:creator>Krom, Y.</dc:creator>
<dc:creator>van der Hoorn, D.</dc:creator>
<dc:creator>Bouwman, L. F.</dc:creator>
<dc:creator>Ritchie, M. E.</dc:creator>
<dc:creator>Reversade, B.</dc:creator>
<dc:creator>Prin, F.</dc:creator>
<dc:creator>Mohun, T.</dc:creator>
<dc:creator>van der Maarel, S. M.</dc:creator>
<dc:creator>McGlinn, E.</dc:creator>
<dc:creator>Murphy, J. M.</dc:creator>
<dc:creator>Keniry, A.</dc:creator>
<dc:creator>de Greef, J. C.</dc:creator>
<dc:creator>Blewitt, M. E.</dc:creator>
<dc:date>2021-05-13</dc:date>
<dc:identifier>doi:10.1101/2021.05.12.443934</dc:identifier>
<dc:title><![CDATA[SMCHD1 has separable roles in chromatin architecture and gene silencing that could be targeted in disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.05.24.444785v1?rss=1">
<title>
<![CDATA[
Automatic instance segmentation of mitochondria in electron microscopy data 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.05.24.444785v1?rss=1"
</link>
<description><![CDATA[
We present a new method for rapid, automated, large-scale 3D mitochondria instance segmentation, developed in response to the ISBI 2021 MitoEM Challenge. In brief, we trained separate machine learning algorithms to predict (1) mitochondria areas and (2) mitochondria boundaries in image volumes acquired from both rat and human cortex with multi-beam scanning electron microscopy. The predictions from these algorithms were combined in a multi-step post-processing procedure, that resulted in high semantic and instance segmentation performance. All code is provided via a public repository.
]]></description>
<dc:creator>Nightingale, L.</dc:creator>
<dc:creator>de Folter, J.</dc:creator>
<dc:creator>Spiers, H.</dc:creator>
<dc:creator>Strange, A.</dc:creator>
<dc:creator>Collinson, L. M.</dc:creator>
<dc:creator>Jones, M. L.</dc:creator>
<dc:date>2021-05-24</dc:date>
<dc:identifier>doi:10.1101/2021.05.24.444785</dc:identifier>
<dc:title><![CDATA[Automatic instance segmentation of mitochondria in electron microscopy data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.05.26.445751v1?rss=1">
<title>
<![CDATA[
Location and function of all Plasmodium kinesins: key roles in parasite proliferation, polarity, and transmission 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.05.26.445751v1?rss=1"
</link>
<description><![CDATA[
Kinesins are microtubule-based motors important in cell division, motility, polarity, and intracellular transport in many eukaryotes. However, they are poorly studied in the divergent eukaryotic pathogens- Plasmodium spp., the causative agents of malaria, which manifest atypical aspects of cell division and plasticity of morphology throughout the lifecycle in both mammalian and mosquito hosts. Here we describe a genome-wide screen of Plasmodium kinesins, revealing diverse subcellular locations and functions in spindle assembly, axoneme formation and cell morphology. Surprisingly, only kinesin-13 is essential for growth in the mammalian host while the other eight kinesins are required during the proliferative and invasive stages of parasite transmission through the mosquito vector. In-depth analyses of kinesin-13 and kinesin-20 revealed functions in microtubule dynamics during apical cell polarity formation, spindle assembly, and axoneme biogenesis. These findings help us to understand the importance of microtubule motors and may be exploited to discover new therapeutic interventions against malaria.
]]></description>
<dc:creator>Zeeshan, M.</dc:creator>
<dc:creator>Rashpa, R.</dc:creator>
<dc:creator>Ferguson, D. J. P.</dc:creator>
<dc:creator>Abel, S.</dc:creator>
<dc:creator>Chahine, Z.</dc:creator>
<dc:creator>Brady, D.</dc:creator>
<dc:creator>Moores, C. A.</dc:creator>
<dc:creator>Le Roch, K. G.</dc:creator>
<dc:creator>Brochet, M.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:creator>Tewari, R.</dc:creator>
<dc:date>2021-05-27</dc:date>
<dc:identifier>doi:10.1101/2021.05.26.445751</dc:identifier>
<dc:title><![CDATA[Location and function of all Plasmodium kinesins: key roles in parasite proliferation, polarity, and transmission]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.05.27.445572v1?rss=1">
<title>
<![CDATA[
Notch controls the cell cycle to define leader versus follower identities during collective cell migration. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.05.27.445572v1?rss=1"
</link>
<description><![CDATA[
Coordination of cell proliferation and migration is fundamental for life, and its dysregulation has catastrophic consequences, as cancer. How cell cycle progression affects migration, and vice-versa, remains largely unknown. We address these questions by combining in silico modelling and in vivo experimentation in the zebrafish Trunk Neural Crest (TNC). TNC migrate collectively, forming chains with a leader cell directing the movement of trailing followers. We show that the acquisition of migratory identity is autonomously controlled by Notch signalling in TNC. High Notch activity defines leaders, while low Notch determines followers. Moreover, cell cycle progression is required for TNC migration and is regulated by Notch. Cells with low Notch activity stay longer in G1 and become followers, while leaders with high Notch activity quickly undergo G1/S transition and remain in S-phase longer. We propose that migratory behaviours are defined through the interaction of Notch signalling and cell cycle progression.
]]></description>
<dc:creator>Alhashem, Z.</dc:creator>
<dc:creator>Feldner-Busztin, D.</dc:creator>
<dc:creator>Revell, C.</dc:creator>
<dc:creator>Alvarez-Garcillan Portillo, M.</dc:creator>
<dc:creator>Richardson, J.</dc:creator>
<dc:creator>Rocha, M.</dc:creator>
<dc:creator>Gauert, A.</dc:creator>
<dc:creator>Corbeaux, T.</dc:creator>
<dc:creator>Prince, V. E.</dc:creator>
<dc:creator>Bentley, K.</dc:creator>
<dc:creator>Linker, C.</dc:creator>
<dc:date>2021-05-27</dc:date>
<dc:identifier>doi:10.1101/2021.05.27.445572</dc:identifier>
<dc:title><![CDATA[Notch controls the cell cycle to define leader versus follower identities during collective cell migration.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.05.29.446196v1?rss=1">
<title>
<![CDATA[
Cell-autonomous generation of the wave pattern within the vertebrate segmentation clock 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.05.29.446196v1?rss=1"
</link>
<description><![CDATA[
Rhythmic and sequential segmentation of the growing vertebrate body relies on the segmentation clock, a multi-cellular oscillating genetic network. The clock is visible as tissue-level kinematic waves of gene expression that travel through the pre-somitic mesoderm (PSM) and arrest at the position of each forming segment. Here we test how this hallmark wave pattern is driven by culturing single maturing PSM cells. We compare their cell-autonomous oscillatory and arrest dynamics to those we observe in the embryo at cellular resolution, finding similarity in the relative slowing of oscillations and arrest in concert with differentiation. This shows that cell-extrinsic signals are not required by the cells to instruct the developmental program underlying the wave pattern. We show that a cell-autonomous timing activity initiates during cell exit from the tailbud, then runs down in the anterior-ward cell flow in the PSM, thereby using elapsed time to provide positional information to the clock. Exogenous FGF lengthens the duration of the cell-intrinsic timer, indicating extrinsic factors in the embryo may regulate the segmentation clock via the timer. In sum, our work suggests that a noisy cell-autonomous, intrinsic timer drives the slowing and arrest of oscillations underlying the wave pattern, while extrinsic factors in the embryo tune this timers duration and precision. This is a new insight into the balance of cell-intrinsic and -extrinsic mechanisms driving tissue patterning in development.
]]></description>
<dc:creator>Rohde, L. A.</dc:creator>
<dc:creator>Bercowsky-Rama, A.</dc:creator>
<dc:creator>Negrete, J.</dc:creator>
<dc:creator>Valentin, G.</dc:creator>
<dc:creator>Naganathan, S. R.</dc:creator>
<dc:creator>Desai, R. A.</dc:creator>
<dc:creator>Strnad, P.</dc:creator>
<dc:creator>Soroldoni, D.</dc:creator>
<dc:creator>Julicher, F.</dc:creator>
<dc:creator>Oates, A. C.</dc:creator>
<dc:date>2021-05-30</dc:date>
<dc:identifier>doi:10.1101/2021.05.29.446196</dc:identifier>
<dc:title><![CDATA[Cell-autonomous generation of the wave pattern within the vertebrate segmentation clock]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.05.29.446277v1?rss=1">
<title>
<![CDATA[
Eosinophils are an integral component of the pulmonary granulocyte response in Tuberculosis and promote host resistance in mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.05.29.446277v1?rss=1"
</link>
<description><![CDATA[
Host resistance to Mycobacterium tuberculosis infection requires the activities of multiple leukocyte subsets, yet the roles of the different innate effector cells during tuberculosis are incompletely understood. Here we uncover an unexpected association between eosinophils and Mtb infection. In humans, eosinophils are decreased in the blood but enriched in resected human tuberculosis lung lesions and autopsy granulomas. Influx of eosinophils is also evident in infected zebrafish, mice, and nonhuman primate granulomas, where they are functionally activated and degranulate. Importantly, employing complementary genetic models of eosinophil deficiency, we demonstrate that, in mice, eosinophils are required for optimal pulmonary bacterial control and host survival after Mtb infection. Collectively, our findings uncover an unexpected recruitment of eosinophils to the infected lung tissue and a protective role for these cells in the control of Mtb infection in mice.
]]></description>
<dc:creator>Bohrer, A. C.</dc:creator>
<dc:creator>Castro, E.</dc:creator>
<dc:creator>Hu, Z.</dc:creator>
<dc:creator>Queiroz, A. T. L.</dc:creator>
<dc:creator>Tocheny, C. E.</dc:creator>
<dc:creator>Assmann, M.</dc:creator>
<dc:creator>Sakai, S.</dc:creator>
<dc:creator>Nelson, C.</dc:creator>
<dc:creator>Baker, P. J.</dc:creator>
<dc:creator>Ma, H.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Zilu, W.</dc:creator>
<dc:creator>du Bruyn, E.</dc:creator>
<dc:creator>Riou, C.</dc:creator>
<dc:creator>Kauffman, K. D.</dc:creator>
<dc:creator>Tuberculosis Imaging Program,</dc:creator>
<dc:creator>Moore, I. N.</dc:creator>
<dc:creator>Del Nonno, F.</dc:creator>
<dc:creator>Petrone, L.</dc:creator>
<dc:creator>Goletti, D.</dc:creator>
<dc:creator>Martineau, A. R. R.</dc:creator>
<dc:creator>Lowe, D. M.</dc:creator>
<dc:creator>Cronan, M. R.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Barry, C. E.</dc:creator>
<dc:creator>Via, L. E.</dc:creator>
<dc:creator>Barber, D. L.</dc:creator>
<dc:creator>Klion, A. D.</dc:creator>
<dc:creator>Andrade, B. B.</dc:creator>
<dc:creator>Song, Y.</dc:creator>
<dc:creator>Wong, K.-W.</dc:creator>
<dc:creator>Mayer-Barber, K. D.</dc:creator>
<dc:date>2021-05-31</dc:date>
<dc:identifier>doi:10.1101/2021.05.29.446277</dc:identifier>
<dc:title><![CDATA[Eosinophils are an integral component of the pulmonary granulocyte response in Tuberculosis and promote host resistance in mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.06.11.447968v1?rss=1">
<title>
<![CDATA[
Principles of ecDNA random inheritance drive rapid genome change and therapy resistance in human cancers 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.06.11.447968v1?rss=1"
</link>
<description><![CDATA[
The foundational principles of Darwinian evolution are variation, selection, and identity by descent. Oncogene amplification on extrachromosomal DNA (ecDNA) is a common event, driving aggressive tumour growth, drug resistance, and shorter survival in patients1-4. Currently, the impact of non-chromosomal oncogene inheritance--random identity by descent--is not well understood. Neither is the impact of ecDNA on variation and selection. Here, integrating mathematical modeling, unbiased image analysis, CRISPR-based ecDNA tagging, and live-cell imaging, we identify a set of basic "rules" for how random ecDNA inheritance drives oncogene copy number and distribution, resulting in extensive intratumoural ecDNA copy number heterogeneity and rapid adaptation to metabolic stress and targeted cancer treatment. Observed ecDNAs obligatorily benefit host cell survival or growth and can change within a single cell cycle. In studies ranging from well-curated, patient-derived cancer cell cultures to clinical tumour samples from patients with glioblastoma and neuroblastoma treated with oncogene-targeted drugs, we show how these ecDNA inheritance "rules" can predict, a priori, some of the aggressive features of ecDNA-containing cancers. These properties are entailed by their ability to rapidly change their genomes in a way that is not possible for cancers driven by chromosomal oncogene amplification. These results shed new light on how the non-chromosomal random inheritance pattern of ecDNA underlies poor outcomes for cancer patients.
]]></description>
<dc:creator>Lange, J. T.</dc:creator>
<dc:creator>Chen, C. Y.</dc:creator>
<dc:creator>Pichugin, Y.</dc:creator>
<dc:creator>Xie, F.</dc:creator>
<dc:creator>Tang, J.</dc:creator>
<dc:creator>Hung, K. L.</dc:creator>
<dc:creator>Yost, K. E.</dc:creator>
<dc:creator>Shi, Q.</dc:creator>
<dc:creator>Erb, M. L.</dc:creator>
<dc:creator>Rajkumar, U.</dc:creator>
<dc:creator>Wu, S.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Liu, Z.</dc:creator>
<dc:creator>Huang, W.</dc:creator>
<dc:creator>Chang, H. Y.</dc:creator>
<dc:creator>Bafna, V.</dc:creator>
<dc:creator>Henssen, A. G.</dc:creator>
<dc:creator>Werner, B.</dc:creator>
<dc:creator>Mischel, P. S.</dc:creator>
<dc:date>2021-06-11</dc:date>
<dc:identifier>doi:10.1101/2021.06.11.447968</dc:identifier>
<dc:title><![CDATA[Principles of ecDNA random inheritance drive rapid genome change and therapy resistance in human cancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.06.15.448466v1?rss=1">
<title>
<![CDATA[
Early mismatch between cancer cells and their tumour microenvironment blocks long term growth 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.06.15.448466v1?rss=1"
</link>
<description><![CDATA[
Cancer cells thrive embedded in a fine-tuned cellular and extracellular environment or tumour microenvironment (TME). There is a general understanding of a co-evolution between cancer cells and their surrounding TME, pointing at a functional connection between cancer cells characteristics and the perturbations induced in their surrounding tissue. However, whether this functional connection needs to be set from the start or if aggressive cancer cells can always be dominating their microenvironment has never been formally proven with a dedicated experimental setting where malignant cells can be challenged to grow in a different TME from the one they would naturally create. Here we generated an experimental setting where we transiently perturb the secretory profile of aggressive breast cancer cells without affecting their intrinsic growth ability. This led to the initial establishment of an atypical TME. Interestingly, even if initially tumours are formed, this atypical TME evolves to impair long term in vivo cancer growth. Using a combination of in vivo transcriptomics, protein arrays and in vitro co-cultures, we found that the atypical TME culminates in the infiltration of macrophages with STAT1high activity. These macrophages show strong anti-tumoural functions which reduce long-term tumour growth, despite lacking canonical M1 markers. Importantly, gene signatures of the mesenchymal compartment of the TME, as well as the anti-tumoural macrophages show striking prognostic power-correlating with less aggressive human breast cancers.
]]></description>
<dc:creator>Perdrix Rosell, A.</dc:creator>
<dc:creator>Maiques, O.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Ombrato, L.</dc:creator>
<dc:creator>Sanz-Moreno, V.</dc:creator>
<dc:creator>Malanchi, I.</dc:creator>
<dc:date>2021-06-15</dc:date>
<dc:identifier>doi:10.1101/2021.06.15.448466</dc:identifier>
<dc:title><![CDATA[Early mismatch between cancer cells and their tumour microenvironment blocks long term growth]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.06.18.448936v1?rss=1">
<title>
<![CDATA[
Structure and dynamics of the chromatin remodeler ALC1 bound to a PARylated nucleosome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.06.18.448936v1?rss=1"
</link>
<description><![CDATA[
The chromatin remodeler ALC1 is recruited to and activated by DNA damage-induced poly(ADP-ribose) (PAR) chains deposited by PARP1/PARP2/HPF1 upon detection of DNA lesions. ALC1 has emerged as a candidate drug target for cancer therapy as its loss confers synthetic lethality in homologous recombination-deficient cells. However, structure-based drug design and molecular analysis of ALC1 have been hindered by the requirement for PARylation and the highly heterogeneous nature of this post-translational modification. Here, we reconstituted an ALC1 and PARylated nucleosome complex modified in vitro using PARP2 and HPF1. This complex was amenable to cryo-EM structure determination without cross-linking, which enabled visualization of several intermediate states of ALC1 from the recognition of the PARylated nucleosome to the tight binding and activation of the remodeler. Functional biochemical assays with PARylated nucleosomes highlight the importance of nucleosomal epitopes for productive remodeling and reveal that ALC1 preferentially slides nucleosomes away from DNA breaks.
]]></description>
<dc:creator>Bacic, L.</dc:creator>
<dc:creator>Gaullier, G.</dc:creator>
<dc:creator>Sabantsev, A.</dc:creator>
<dc:creator>Lehmann, L.</dc:creator>
<dc:creator>Brackmann, K.</dc:creator>
<dc:creator>Dimakou, D.</dc:creator>
<dc:creator>Halic, M.</dc:creator>
<dc:creator>Hewitt, G.</dc:creator>
<dc:creator>Boulton, S. J.</dc:creator>
<dc:creator>Deindl, S.</dc:creator>
<dc:date>2021-06-18</dc:date>
<dc:identifier>doi:10.1101/2021.06.18.448936</dc:identifier>
<dc:title><![CDATA[Structure and dynamics of the chromatin remodeler ALC1 bound to a PARylated nucleosome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.04.20.050674v1?rss=1">
<title>
<![CDATA[
CDK1 controls CHMP7-dependent nuclear envelope reformation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.04.20.050674v1?rss=1"
</link>
<description><![CDATA[
Through membrane sealing and disassembly of spindle microtubules, the Endosomal Sorting Complex Required for Transport-III (ESCRT-III) machinery has emerged as a key player in the regeneration of a sealed nuclear envelope (NE) during mitotic exit, and in the repair of this organelle during interphase rupture. ESCRT-III assembly at the NE occurs transiently during mitotic exit and is initiated when CHMP7, an ER-localised ESCRT-II/ESCRT-III hybrid protein, interacts with the Inner Nuclear Membrane (INM) protein LEM2. Whilst classical nucleocytoplasmic transport mechanisms have been proposed to separate LEM2 and CHMP7 during interphase, it is unclear how CHMP7 assembly is suppressed in mitosis when NE and ER identities are mixed. Here, we use live cell imaging and protein biochemistry to examine the biology of these proteins during mitotic exit. Firstly, we show that CHMP7 plays an important role in the dissolution of LEM2 clusters that form at the NE during M-exit. Secondly, we show that CDK1 phosphorylates CHMP7 upon mitotic entry at Ser3 and Ser441 and that this phosphorylation suppresses CHMP7s interaction with LEM2, limiting its assembly during M-phase. We show that spatiotemporal differences in the dephosphorylation of CHMP7 license its assembly at the NE during telophase, but restrict its assembly on the ER at this time. Without CDK1 phosphorylation, CHMP7 undergoes inappropriate assembly in the peripheral ER during M-exit, capturing LEM2 and downstream ESCRT-III components. Lastly, we establish that a microtubule network is dispensable for ESCRT-III assembly at the reforming nuclear envelope. These data identify a key cell-cycle control programme allowing ESCRT-III-dependent nuclear regeneration.
]]></description>
<dc:creator>Gatta, A. T.</dc:creator>
<dc:creator>Olmos, Y.</dc:creator>
<dc:creator>Stoten, C. L.</dc:creator>
<dc:creator>Carlton, J. G.</dc:creator>
<dc:date>2020-04-21</dc:date>
<dc:identifier>doi:10.1101/2020.04.20.050674</dc:identifier>
<dc:title><![CDATA[CDK1 controls CHMP7-dependent nuclear envelope reformation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.06.23.449503v1?rss=1">
<title>
<![CDATA[
LAP1 regulates nuclear plasticity to enable constrained migration 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.06.23.449503v1?rss=1"
</link>
<description><![CDATA[
Metastasis involves dissemination of cancer cells away from a primary tumour and colonisation at distal sites. During this process, cancer cells must negotiate multiple physical constraints imposed by the microenvironment and tissue structure. The biophysical properties of the nucleus must be tuned since they pose a challenge to constrained migration. By analysing nuclear genes upregulated during the acquisition of metastatic potential, we discovered increased expression of the inner nuclear membrane protein LAP1 in metastatic melanoma cells and at the invasive fronts of human primary tumours and in metastases. Human cells express two LAP1 isoforms (LAP1B and LAP1C), which differ in their amino terminus. We found that whereas the longer isoform, LAP1B, binds more strongly to nuclear lamins and has restricted motility within the nuclear envelope, the shorter isoform, LAP1C, favours nuclear envelope blebbing and allows migration through constraints. We propose that LAP1 renders the nucleus plastic and enhances melanoma aggressiveness.
]]></description>
<dc:creator>Jung Garcia, Y.</dc:creator>
<dc:creator>Maiques, O.</dc:creator>
<dc:creator>Rodriguez-Hernandez, I.</dc:creator>
<dc:creator>Fanshawe, B.</dc:creator>
<dc:creator>Domart, M.-C.</dc:creator>
<dc:creator>Renshaw, M. J.</dc:creator>
<dc:creator>Marti, R. M.</dc:creator>
<dc:creator>Matias-Guiu, X.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:creator>Sanz-Moreno, V.</dc:creator>
<dc:creator>Carlton, J. G.</dc:creator>
<dc:date>2021-06-23</dc:date>
<dc:identifier>doi:10.1101/2021.06.23.449503</dc:identifier>
<dc:title><![CDATA[LAP1 regulates nuclear plasticity to enable constrained migration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.06.28.450144v1?rss=1">
<title>
<![CDATA[
Sparse connectivity for MAP inference in linear models using sister mitral cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.06.28.450144v1?rss=1"
</link>
<description><![CDATA[
Sensory processing is hard because the variables of interest are encoded in spike trains in a relatively complex way. A major goal in studies of sensory processing is to understand how the brain extracts those variables. Here we revisit a common encoding model in which variables are encoded linearly. Although there are typically more variables than neurons, this problem is still solvable because only a small number of variables appear at any one time (sparse prior). However, previous solutions require all-to-all connectivity, inconsistent with the sparse connectivity seen in the brain. Here we propose an algorithm that provably reaches the MAP (maximum a posteriori) inference solution, but does so using sparse connectivity. Our algorithm is inspired by the circuit of the mouse olfactory bulb, but our approach is general enough to apply to other modalities. In addition, it should be possible to extend it to nonlinear encoding models.

SummarySensory systems must infer latent variables from noisy and ambiguous input. MAP inference - choosing the most likely latent variable given the sensory input - is one of the simplest methods for doing that, but its neural implementation often requires all-to-all connectivity between the neurons involved. In common sensory contexts this can require a single neuron to connect to hundreds of thousands of others, which is biologically implausible. In this work we take inspiration from the  sister mitral cells of the olfactory system - groups of neurons associated with the same input channel - to derive a method for performing MAP inference using sparse connectivity. We do so by assigning sister cells to random subsets of the latent variables and using additional cells to ensure that sisters correctly share information. We then derive the circuitry and dynamics required for the sister cells to compute the original MAP inference solution. Our work yields a biologically plausible circuit that provably solves the MAP inference problem and provides experimentally testable predictions. While inspired by the olfactory system, our method is quite general, and is likely to apply to other sensory modalities.
]]></description>
<dc:creator>Tootoonian, S.</dc:creator>
<dc:creator>Schaefer, A. T.</dc:creator>
<dc:creator>Latham, P. E.</dc:creator>
<dc:date>2021-06-28</dc:date>
<dc:identifier>doi:10.1101/2021.06.28.450144</dc:identifier>
<dc:title><![CDATA[Sparse connectivity for MAP inference in linear models using sister mitral cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.06.28.450158v1?rss=1">
<title>
<![CDATA[
Global analysis of putative phospholipases in the malaria parasite  Plasmodium falciparum  reveals critical factors for parasite proliferation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.06.28.450158v1?rss=1"
</link>
<description><![CDATA[
For its replication within red blood cells, the malaria parasite is highly dependent on correctly regulated lipid metabolism. Enzymes involved in lipid metabolic processes are therefore potential drug targets. We here provide a functional analysis of the 20 putative phospholipases that are expressed by asexual blood stages of Plasmodium falciparum. We reveal a high level of redundancy among members of this group, but using conditional mislocalization and gene disruption techniques we show that the phosphoinositide-specific phospholipase C (PF3D7_1013500) has a previously unrecognized essential role in intracellular parasite maturation. In addition, we demonstrate that the patatin-like phospholipase PF3D7_1358000 localizes to the mitochondrion. Parasites lacking this enzyme display a severe growth phenotype and defects in mitochondrial morphogenesis and function leading to hypersensitivity towards proguanil and inhibitors of the mitochondrial electron transport chain including atovaquone. This demonstrates that regulated mitochondrial lipid homeostasis is necessary for mitochondrial function and coordinated division during parasite multiplication.
]]></description>
<dc:creator>Burda, P.-C.</dc:creator>
<dc:creator>Ramaprasad, A.</dc:creator>
<dc:creator>Pietsch, E.</dc:creator>
<dc:creator>Bielfeld, S.</dc:creator>
<dc:creator>Söhnchen, C.</dc:creator>
<dc:creator>Wilcke, L.</dc:creator>
<dc:creator>Strauss, J.</dc:creator>
<dc:creator>Schwudke, D.</dc:creator>
<dc:creator>Sait, A.</dc:creator>
<dc:creator>Collinson, L. M.</dc:creator>
<dc:creator>Blackman, M. J.</dc:creator>
<dc:creator>Gilberger, T.-W.</dc:creator>
<dc:date>2021-06-28</dc:date>
<dc:identifier>doi:10.1101/2021.06.28.450158</dc:identifier>
<dc:title><![CDATA[Global analysis of putative phospholipases in the malaria parasite  Plasmodium falciparum  reveals critical factors for parasite proliferation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.06.30.450614v1?rss=1">
<title>
<![CDATA[
Exploitation of the Secretory Pathway by SARS-CoV-2 Envelope 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.06.30.450614v1?rss=1"
</link>
<description><![CDATA[
The {beta}-coronavirus SARS-CoV-2 is the causative agent of the global Covid-19 pandemic. Coronaviral Envelope (E) proteins are pentameric viroporins that play essential roles in assembly, release and pathogenesis. We developed an inert tagging strategy for SARS-CoV-2 E and find that it localises to the Golgi and to lysosomes. We identify sequences in E, conserved across Coronaviridae, responsible for ER-to-Golgi export, and relate this activity to interaction with COP-II via SEC24. Using proximity biotinylation, we identify host-cell factors that interact with E and identify an ARFRP1/AP-1 dependent pathway allowing Golgi-to-lysosome trafficking of E. We identify sequences in E that bind AP-1, are conserved across {beta}-coronaviruses and allow E to be trafficked from Golgi to lysosomes. We show that E acts to deacidify lysosomes and by developing a trans-complementation assay, we show that both lysosomal trafficking of E and its viroporin activity are necessary for efficient viral replication and release.
]]></description>
<dc:creator>Pearson, G. J.</dc:creator>
<dc:creator>Broncel, M.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Carlton, J. G.</dc:creator>
<dc:date>2021-06-30</dc:date>
<dc:identifier>doi:10.1101/2021.06.30.450614</dc:identifier>
<dc:title><![CDATA[Exploitation of the Secretory Pathway by SARS-CoV-2 Envelope]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.06.30.450572v1?rss=1">
<title>
<![CDATA[
Functional profiling of long intergenic non-coding RNAs in fission yeast 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.06.30.450572v1?rss=1"
</link>
<description><![CDATA[
Eukaryotic genomes express numerous long intergenic non-coding RNAs (lincRNAs) that do not overlap any coding genes. Some lincRNAs function in various aspects of gene regulation, but it is not clear in general to what extent lincRNAs contribute to the information flow from genotype to phenotype. To explore this question, we systematically analyzed cellular roles of lincRNAs in Schizosaccharomyces pombe. Using seamless CRISPR/Cas9-based genome editing, we deleted 141 lincRNA genes to broadly phenotype these mutants, together with 238 diverse coding-gene mutants for functional context. We applied high-throughput colony-based assays to determine mutant growth and viability in benign conditions and in response to 145 different nutrient, drug and stress conditions. These analyses uncovered phenotypes for 47.5% of the lincRNAs and 96% of the protein-coding genes. For 110 lincRNA mutants, we also performed high-throughput microscopy and flow-cytometry assays, linking 37% of these lincRNAs with cell-size and/or cell-cycle control. With all assays combined, we detected phenotypes for 84 (59.6%) of all lincRNA deletion mutants tested. For complementary functional inference, we analyzed colony growth of strains ectopically overexpressing 113 lincRNA genes under 47 different conditions. Of these overexpression strains, 102 (90.3%) showed altered growth under certain conditions. Clustering analyses provided further functional clues and relationships for some of the lincRNAs. These rich phenomics datasets associate lincRNA mutants with hundreds of phenotypes, indicating that most of the lincRNAs analyzed exert cellular functions in specific environmental or physiological contexts. This study provides groundwork to further dissect the roles of these lincRNAs in the relevant conditions.
]]></description>
<dc:creator>Rodriguez-Lopez, M.</dc:creator>
<dc:creator>Anver, S.</dc:creator>
<dc:creator>Cotobal, C.</dc:creator>
<dc:creator>Kamrad, S.</dc:creator>
<dc:creator>Malecki, M.</dc:creator>
<dc:creator>Correia-Melo, C.</dc:creator>
<dc:creator>Hoti, M.</dc:creator>
<dc:creator>Townsend, S.</dc:creator>
<dc:creator>Marguerat, S. B.</dc:creator>
<dc:creator>Pong, S.</dc:creator>
<dc:creator>Wu, M.</dc:creator>
<dc:creator>Montemayor, L.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:creator>Bahler, J.</dc:creator>
<dc:date>2021-07-01</dc:date>
<dc:identifier>doi:10.1101/2021.06.30.450572</dc:identifier>
<dc:title><![CDATA[Functional profiling of long intergenic non-coding RNAs in fission yeast]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.07.06.451052v1?rss=1">
<title>
<![CDATA[
Isoform resolved measurements of absolute translational efficiency define interplay of HIF and mTOR dysregulation in kidney cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.07.06.451052v1?rss=1"
</link>
<description><![CDATA[
Hypoxia inducible factor (HIF) and mammalian target of rapamycin (mTOR) pathways orchestrate responses to oxygen and nutrient availability. These pathways are frequently dysregulated in cancer, but their interplay is poorly understood, in part because of difficulties in simultaneous measurement of global and mRNA-specific translation. Here we describe a workflow for measurement of absolute translational efficiency of mRNAs resolved by their transcription start sites (TSSs). Its application to kidney cancer cells revealed a remarkable extent of translational reprogramming by mTOR, strongly affecting many metabolic enzymes and pathways. By contrast, global effects of HIF on translation were limited, and we did not observe reported translational activation by HIF2A. In contrast, HIF-dependent alterations in TSS usage were associated with robust changes in translational efficiency in a subset of genes. Analyses of the interplay of HIF and mTOR revealed that specific classes of HIF1A and HIF2A transcriptional target gene manifest markedly different sensitivity to mTOR, in a manner that supports combined use of HIF2A and mTOR inhibitors in treatment of kidney cancer.
]]></description>
<dc:creator>Sugimoto, Y.</dc:creator>
<dc:creator>Ratcliffe, P. J.</dc:creator>
<dc:date>2021-07-06</dc:date>
<dc:identifier>doi:10.1101/2021.07.06.451052</dc:identifier>
<dc:title><![CDATA[Isoform resolved measurements of absolute translational efficiency define interplay of HIF and mTOR dysregulation in kidney cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.07.10.451887v1?rss=1">
<title>
<![CDATA[
TGFβ signalling is required to maintain pluripotency of human naïve pluripotent stem cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.07.10.451887v1?rss=1"
</link>
<description><![CDATA[
The signalling pathways that maintain primed human pluripotent stem cells (hPSCs) have been well characterised, revealing a critical role for TGF{beta}/Activin/Nodal signalling. In contrast, the signalling requirements of naive human pluripotency have not been fully established. Here, we demonstrate that TGF{beta} signalling is required to maintain naive hPSCs. The downstream effector proteins - SMAD2/3 - bind common sites in naive and primed hPSCs, including shared pluripotency genes. In naive hPSCs, SMAD2/3 additionally bind to active regulatory regions near to naive pluripotency genes. Inhibiting TGF{beta} signalling in naive hPSCs causes the downregulation of SMAD2/3-target genes and pluripotency exit. Single-cell analyses reveal that naive and primed hPSCs follow different transcriptional trajectories after inhibition of TGF{beta} signalling. Primed hPSCs differentiate into neuroectoderm cells, whereas naive hPSCs transition into trophectoderm. These results establish that there is a continuum for TGF{beta} pathway function in human pluripotency spanning a developmental window from naive to primed states.
]]></description>
<dc:creator>Osnato, A.</dc:creator>
<dc:creator>Brown, S.</dc:creator>
<dc:creator>Krueger, C.</dc:creator>
<dc:creator>Andrews, S.</dc:creator>
<dc:creator>Collier, A. J.</dc:creator>
<dc:creator>Nakanoh, S.</dc:creator>
<dc:creator>Quiroga Londono, M.</dc:creator>
<dc:creator>Wesley, B. T.</dc:creator>
<dc:creator>Muraro, D.</dc:creator>
<dc:creator>Brumm, S.</dc:creator>
<dc:creator>Niakan, K.</dc:creator>
<dc:creator>Vallier, L.</dc:creator>
<dc:creator>Ortmann, D.</dc:creator>
<dc:creator>Rugg-Gunn, P. J.</dc:creator>
<dc:date>2021-07-11</dc:date>
<dc:identifier>doi:10.1101/2021.07.10.451887</dc:identifier>
<dc:title><![CDATA[TGFβ signalling is required to maintain pluripotency of human naïve pluripotent stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.07.12.452027v1?rss=1">
<title>
<![CDATA[
Immunogenicity and pre-clinical efficacy of an OMV-based SARS-CoV-2 vaccine 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.07.12.452027v1?rss=1"
</link>
<description><![CDATA[
The vaccination campaign against SARS-CoV-2 relies on the world-wide availability of effective vaccines, with a potential need of 20 billion vaccine doses to fully vaccinate the world population. To reach this goal, the manufacturing and logistic processes should be affordable to all countries, irrespectively of economical and climatic conditions.

Outer membrane vesicles (OMVs) are bacterial-derived vesicles that can be engineered to incorporate heterologous antigens. Given the inherent adjuvanticity, such modified OMVs can be used as vaccine to induce potent immune responses against the associated protein. Here we show that OMVs engineered to incorporate peptides derived from the receptor binding motif (RBM) of the spike protein from SARS-CoV-2 elicit an effective immune response in immunized mice, resulting in the production of neutralizing antibodies. The immunity induced by the vaccine is sufficient to protect K18-hACE2 transgenic mice from intranasal challenge with SARS-CoV-2, preventing both virus replication in the lungs and the pathology associated with virus infection. Furthermore, we show that OMVs can be effectively decorated with RBM peptides derived from a different genetic variant of SARS-CoV-2, inducing a similarly potent neutralization activity in vaccinated mice. Altogether, given the convenience associated with ease of engineering, production and distribution, our results demonstrate that OMV-based SARS-CoV-2 vaccines can be a crucial addition to the vaccines currently available.
]]></description>
<dc:creator>Grandi, A.</dc:creator>
<dc:creator>Tomasi, M.</dc:creator>
<dc:creator>Accordini, S.</dc:creator>
<dc:creator>Bertelli, C.</dc:creator>
<dc:creator>Vanzo, T.</dc:creator>
<dc:creator>Gagliardi, A.</dc:creator>
<dc:creator>Caproni, E.</dc:creator>
<dc:creator>Tamburini, S.</dc:creator>
<dc:creator>Fantappie, L.</dc:creator>
<dc:creator>Di Lascio, G.</dc:creator>
<dc:creator>Bisoffi, Z.</dc:creator>
<dc:creator>Piubelli, C.</dc:creator>
<dc:creator>Valenti, M. T.</dc:creator>
<dc:creator>Dalle Carbonare, L.</dc:creator>
<dc:creator>Zipeto, D.</dc:creator>
<dc:creator>Rava, M.</dc:creator>
<dc:creator>Fumagalli, V.</dc:creator>
<dc:creator>Di Lucia, P.</dc:creator>
<dc:creator>Marotta, D.</dc:creator>
<dc:creator>Sala, E.</dc:creator>
<dc:creator>Iannacone, M.</dc:creator>
<dc:creator>Cherepanov, P.</dc:creator>
<dc:creator>Bolognesi, M.</dc:creator>
<dc:creator>Pizzato, M.</dc:creator>
<dc:creator>Grandi, G.</dc:creator>
<dc:date>2021-07-13</dc:date>
<dc:identifier>doi:10.1101/2021.07.12.452027</dc:identifier>
<dc:title><![CDATA[Immunogenicity and pre-clinical efficacy of an OMV-based SARS-CoV-2 vaccine]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.07.11.451940v1?rss=1">
<title>
<![CDATA[
Suppressed eusocial reproduction supports evolutionary convergence over co-option 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.07.11.451940v1?rss=1"
</link>
<description><![CDATA[
Eusociality represents a major evolutionary transition that arose independently in at least 12 insect lineages. Despite this prevalence, there remains considerable uncertainty surrounding the catalysing event and underlying genomic changes that enable such modifications. Commonly associated with this evolutionary transition is establishing and maintaining the reproductive division of labour (e.g. a reproductive queen and no-reproductive workers). This division is, at least in part, induced and maintained by highly species-specific pheromones. However, genomic analysis remains conflicted on the role of pheromones in this evolutionary transition. Specifically, if there was co-option of a common pheromone-sensitive genetic pathway present in all progenitor species or strong lineage-specific selection converging on similar transcriptomic signatures.

Using a solitary insect model, we sought to determine if various species-specific pheromones induced similar transcriptomic responses, thus activating similar pathways. We measured the transcriptomic and physiological response of a solitary insect, Drosophila melanogaster, to pheromones from bumblebees, honey bees, and termites. Each treatment induced the same strong physiological response - a decreased ovary size. However, employing several methods of transcriptomic analysis, we did not observe conservation in pheromone-mediated gene/pathway regulation.

Thus, despite a conserved phenotypic response, the underpinning transcriptome was vastly different. This suggests that pheromone-mediated eusociality is the result of convergent evolution. We propose that mechanisms maintaining eusociality (i.e. proto-pheromone) in early stages of eusocial evolution in each group, thus, acting as a primer for eusociality. This early state is then refined through strong selective pressure, resulting in a converging eusocial phenotype.

Visual Abstract

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View larger version (32K):
org.highwire.dtl.DTLVardef@117ad33org.highwire.dtl.DTLVardef@16bf4f5org.highwire.dtl.DTLVardef@9f215aorg.highwire.dtl.DTLVardef@146c0f5_HPS_FORMAT_FIGEXP  M_FIG O_FLOATNOFigure 1.C_FLOATNO C_FIG
]]></description>
<dc:creator>Velasque, M.</dc:creator>
<dc:creator>Yongkai, T.</dc:creator>
<dc:creator>Liu, A. W.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:creator>Denton, J. A.</dc:creator>
<dc:date>2021-07-12</dc:date>
<dc:identifier>doi:10.1101/2021.07.11.451940</dc:identifier>
<dc:title><![CDATA[Suppressed eusocial reproduction supports evolutionary convergence over co-option]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.07.14.451473v1?rss=1">
<title>
<![CDATA[
Ribocutter: Cas9-mediated rRNA depletion from multiplexed riboseq libraries 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.07.14.451473v1?rss=1"
</link>
<description><![CDATA[
RNA sequencing libraries produced from fragmented RNA, especially Ribo-seq libraries, contain high proportions of reads from abundant non-coding RNAs. Here, we describe a streamlined Cas9-based protocol for removing abundant rRNA/ncRNA contaminants from Ribo-seq (or other small RNA-seq) libraries and an easy-to-use software tool, ribocutter, for designing ready-to-order sgRNA templates. Following sgRNA template design, the pool of templates is in vitro transcribed using a 1-step commercial kit, which produces enough sgRNAs for multiple treatments. A single multiplexed sequencing library is then treated with Cas9/sgRNAs, followed by a short PCR program, which can increase the fraction of useful reads by more than 3-fold. Comparison of samples before and after depletion demonstrates that Cas9 produces minimal off-target effects and preserves key features (eg. footprint length, periodicity) of Ribo-seq libraries. The method is thus highly effective, costs <{pound}0.50 per sample, and minimises non-specific depletion and technical variation between samples.
]]></description>
<dc:creator>Wilkins, O. G.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:date>2021-07-14</dc:date>
<dc:identifier>doi:10.1101/2021.07.14.451473</dc:identifier>
<dc:title><![CDATA[Ribocutter: Cas9-mediated rRNA depletion from multiplexed riboseq libraries]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.07.12.452033v1?rss=1">
<title>
<![CDATA[
Flight-tone mediated circadian control of audibility in mating swarms of Anopheles mosquitoes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.07.12.452033v1?rss=1"
</link>
<description><![CDATA[
Mating swarms of malaria mosquitoes form every day at sunset throughout the tropical world, they typically last less than 30 minutes. Activity patterns must thus be highly synchronized between the sexes. Moreover, males must be able to identify the few sporadically entering females by detecting the females faint flight tones. We here show that the Anopheles circadian clock ensures a tight synchrony of male and female activity and -importantly - also retunes the males acoustic detection system: by lifting their own flight tones at dusk, males actively enhance the audibility of females. The reported phenomenon of  harmonic convergence is a random by-product of the mosquitoes flight tone variance. Intriguingly, flight tones of individual mosquitoes occupy narrow -partly non-overlapping-frequency ranges, suggesting that the audibility of individual females varies across males.

One Sentence SummaryMale mosquitoes sharpen their hearing at sunset.
]]></description>
<dc:creator>Somers, J.</dc:creator>
<dc:creator>Georgiades, M.</dc:creator>
<dc:creator>Su, M. P.</dc:creator>
<dc:creator>Bagi, J.</dc:creator>
<dc:creator>Andres, M.</dc:creator>
<dc:creator>Mills, G.</dc:creator>
<dc:creator>Ntabaliba, W.</dc:creator>
<dc:creator>Moore, S. J.</dc:creator>
<dc:creator>Spaccapelo, R.</dc:creator>
<dc:creator>Albert, J. T.</dc:creator>
<dc:date>2021-07-12</dc:date>
<dc:identifier>doi:10.1101/2021.07.12.452033</dc:identifier>
<dc:title><![CDATA[Flight-tone mediated circadian control of audibility in mating swarms of Anopheles mosquitoes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.07.18.452838v1?rss=1">
<title>
<![CDATA[
TAF4b transcription networks regulating early oocyte differentiation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.07.18.452838v1?rss=1"
</link>
<description><![CDATA[
Establishment of a healthy ovarian reserve is contingent upon numerous regulatory pathways during embryogenesis. Previously, mice lacking TBP-associated factor 4b (Taf4b) were shown to exhibit a diminished ovarian reserve. However, potential oocyte-intrinsic functions of TAF4b have not been examined. Here we use a combination of gene expression profiling and chromatin mapping to characterize the TAF4b gene regulatory network in mouse oocytes. We find that Taf4b-deficient oocytes display inappropriate expression of meiotic, chromatin, and X-linked genes, and unexpectedly we found a connection with Turner Syndrome pathways. Using Cleavage Under Targets and Release Using Nuclease (CUT&RUN), we observed TAF4b enrichment at genes involved in meiosis and DNA repair, some of which are differentially expressed in Taf4b-deficient oocytes. Interestingly, TAF4b target genes were enriched for Sp/KLF family motifs rather than TATA-box, suggesting an alternate mode of promoter interaction. Together, our data connects several gene regulatory nodes that contribute to the ovarian reserve.
]]></description>
<dc:creator>Gura, M. A.</dc:creator>
<dc:creator>Relovska, S.</dc:creator>
<dc:creator>Abt, K. M.</dc:creator>
<dc:creator>Seymour, K. A.</dc:creator>
<dc:creator>Wu, T.</dc:creator>
<dc:creator>Kaya, H.</dc:creator>
<dc:creator>Turner, J.</dc:creator>
<dc:creator>Fazzio, T. G.</dc:creator>
<dc:creator>Freiman, R. N.</dc:creator>
<dc:date>2021-07-18</dc:date>
<dc:identifier>doi:10.1101/2021.07.18.452838</dc:identifier>
<dc:title><![CDATA[TAF4b transcription networks regulating early oocyte differentiation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.07.20.453046v1?rss=1">
<title>
<![CDATA[
Metabolic turnover and dynamics of modified ribonucleosides by 13C labeling 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.07.20.453046v1?rss=1"
</link>
<description><![CDATA[
Tandem mass spectrometry (MS/MS) is an accurate tool to assess modified ribonucleosides and their dynamics in mammalian cells. Yet, MS/MS quantification of lowly abundant modifications in non-ribosomal RNAs is unreliable, and the dynamic features of various modifications poorly understood. We developed a 13C labeling approach, 13C-dynamods, to quantify the turnover of base modifications in newly transcribed RNA. This turnover-based approach helped to resolve mRNA from ncRNA modifications in purified RNA or free ribonucleosides, and showed the distinct kinetics of N6-methyladenosine (m62A) versus 7-methylguanosine (m7G) in polyA+-purified RNA. We uncovered that N6,N6-dimethyladenosine (m62A) exhibits a distinct turnover in small RNAs and free ribonucleosides when compared to the known m6 A-modified large rRNAs. Finally, combined measurements of turnover and abundance informed on the transcriptional versus posttranscriptional sensitivity of modified ncRNAs and mRNAs, respectively, to stress conditions. Thus, 13C-dynamods enables studies of origin of modified RNAs at steady-state and their dynamics under non-stationary conditions.
]]></description>
<dc:creator>Gameiro, P. A.</dc:creator>
<dc:creator>Encheva, V.</dc:creator>
<dc:creator>Silva dos Santos, M.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:date>2021-07-20</dc:date>
<dc:identifier>doi:10.1101/2021.07.20.453046</dc:identifier>
<dc:title><![CDATA[Metabolic turnover and dynamics of modified ribonucleosides by 13C labeling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.07.25.453647v1?rss=1">
<title>
<![CDATA[
Oncogenic c-Myc induces replication stress by increasing cohesins chromatin occupancy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.07.25.453647v1?rss=1"
</link>
<description><![CDATA[
Oncogene-induced replication stress is a major driver of genomic instability in cancer cells, with a central role in both cancer initiation and progression (1). Despite its critical role in cancer development, the mechanisms that lay at the basis of oncogene-induced replication stress remains poorly understood. Here, we investigate the mechanism of c-Myc-induced replication stress. Our data shows that c-Myc induces replication stress by increasing the amount of cohesins bound to chromatin in the G1 phase of the cell cycle. This is independent of previously suggested mechanisms involving deregulation of replication initiation and transcriptional interference. Restoring the amount of chromatin-bound cohesins to control levels, or preventing the accumulation of cohesins at CTCF sites, in cells experiencing oncogenic c-Myc activity prevents replication stress. Increased cohesins chromatin occupancy correlates with a c-Myc-dependent increase in the levels of the cohesion loader Mau2. Preventing c-Myc-induced increase in Mau2 reduces oncogene-induced replication stress. Together our data support a novel mechanism for oncogene-induced replication stress. Since c-Myc activation is a crucial event in many human cancers (2), identifying the mechanisms through which this oncogene promotes replication stress provides critical insights into cancer biology.
]]></description>
<dc:creator>Peripolli, S.</dc:creator>
<dc:creator>Singh, T.</dc:creator>
<dc:creator>Patel, H.</dc:creator>
<dc:creator>Meneguello, L.</dc:creator>
<dc:creator>Kiso, K.</dc:creator>
<dc:creator>Thorpe, P. H.</dc:creator>
<dc:creator>Bertoli, C.</dc:creator>
<dc:creator>de Bruin, R. A. M.</dc:creator>
<dc:date>2021-07-26</dc:date>
<dc:identifier>doi:10.1101/2021.07.25.453647</dc:identifier>
<dc:title><![CDATA[Oncogenic c-Myc induces replication stress by increasing cohesins chromatin occupancy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.07.28.454135v1?rss=1">
<title>
<![CDATA[
Cell-specific Bioorthogonal Tagging of Glycoproteins in Co-culture 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.07.28.454135v1?rss=1"
</link>
<description><![CDATA[
Altered glycosylation is an undisputed corollary of cancer development. Understanding these alterations is paramount but hampered by limitations underlying cellular model systems. For instance, the intricate interactions between tumour and host cannot be adequately recapitulated in monoculture of tumour-derived cell lines. More complex co-culture models usually rely on sorting procedures for proteome analyses and rarely capture the details of protein glycosylation. Here, we report a strategy termed Bio-Orthogonal Cell line-specific Tagging of Glycoproteins (BOCTAG). Cells are equipped by transfection with an artificial biosynthetic pathway that transforms bioorthogonally tagged sugars into the corresponding nucleotide-sugars. Only transfected cells incorporate bioorthogonal tags into glycoproteins in the presence of non-transfected cells. We employ BOCTAG as an imaging technique and to annotate cell-specific glycosylation sites in mass spectrometry-glycoproteomics. We demonstrate application in co-culture and mouse models, allowing for profiling of the glycoproteome as an important modulator of cellular function.
]]></description>
<dc:creator>Cioce, A.</dc:creator>
<dc:creator>Calle, B.</dc:creator>
<dc:creator>Marchesi, A.</dc:creator>
<dc:creator>Bineva-Todd, G.</dc:creator>
<dc:creator>Flynn, H.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Tastan, O. Y.</dc:creator>
<dc:creator>Roustan, C.</dc:creator>
<dc:creator>Keenan, T.</dc:creator>
<dc:creator>Both, P.</dc:creator>
<dc:creator>Huang, K.</dc:creator>
<dc:creator>Parmeggiani, F.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Kjaer, S.</dc:creator>
<dc:creator>Fascione, M. A.</dc:creator>
<dc:creator>Flitsch, S. L.</dc:creator>
<dc:creator>Schumann, B.</dc:creator>
<dc:date>2021-07-28</dc:date>
<dc:identifier>doi:10.1101/2021.07.28.454135</dc:identifier>
<dc:title><![CDATA[Cell-specific Bioorthogonal Tagging of Glycoproteins in Co-culture]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.08.09.455510v1?rss=1">
<title>
<![CDATA[
SIGNR1 promotes immune dysfunction in systemic candidiasis by modulating neutrophil lifespan via T cell-derived histones and G-CSF 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.08.09.455510v1?rss=1"
</link>
<description><![CDATA[
The mechanisms regulating immune dysfunction during sepsis are poorly understood. Here, we show that neutrophil-derived myeloperoxidase delays the onset of immune dysfunction during systemic candidiasis by controlling microbes captured by splenic marginal zone (MZ) macrophages. In contrast, SIGNR1-mediated microbe capture accelerates MZ colonization and immune dysfunction by triggering T cell death, T cell-dependent chromatin release and the synergistic induction of G-CSF by histones and fungi. Histones and G-CSF promote the prevalence of immature Ly6Glow neutrophils with defective oxidative burst, by selectively shortening the lifespan of mature Ly6Ghigh neutrophils. Consistently, T cell deficiency, or blocking SIGNR1, G-CSF or histones delayed neutrophil dysfunction. Furthermore, histones and G-CSF in the plasma of sepsis patients, shortened neutrophil lifespan and correlated with neutrophil mortality markers associated with a poor prognosis. Hence, the compromise of internal antimicrobial barrier sites drives neutrophil dysfunction by selectively modulating neutrophil lifespan via pathogenic T cell death, extracellular histones, and G-CSF.
]]></description>
<dc:creator>Ioannou, M.</dc:creator>
<dc:creator>Hoving, D.</dc:creator>
<dc:creator>Aramburu, I. V.</dc:creator>
<dc:creator>De Vasconcelos, N. M.</dc:creator>
<dc:creator>Temkin, M. I.</dc:creator>
<dc:creator>Wang, Q.</dc:creator>
<dc:creator>Vernardis, S.</dc:creator>
<dc:creator>Demichev, V.</dc:creator>
<dc:creator>Tsourouktsoglou, T.-D.</dc:creator>
<dc:creator>Boeing, S.</dc:creator>
<dc:creator>Goldstone, R.</dc:creator>
<dc:creator>David, S.</dc:creator>
<dc:creator>Klaus Stahl, K.</dc:creator>
<dc:creator>Bode, C.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:creator>Papagiannopoulos, V.</dc:creator>
<dc:date>2021-08-09</dc:date>
<dc:identifier>doi:10.1101/2021.08.09.455510</dc:identifier>
<dc:title><![CDATA[SIGNR1 promotes immune dysfunction in systemic candidiasis by modulating neutrophil lifespan via T cell-derived histones and G-CSF]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.08.13.456308v1?rss=1">
<title>
<![CDATA[
Structural transitions in the GTP cap visualized by cryo-EM of catalytically inactive microtubules 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.08.13.456308v1?rss=1"
</link>
<description><![CDATA[
Microtubules (MTs) are polymers of /{beta}-tubulin heterodimers that stochastically switch between growth and shrinkage phases. This dynamic instability is critically important for MT function. It is believed that GTP hydrolysis within the MT lattice is accompanied by destabilizing conformational changes, and that MT stability depends on a transiently existing GTP cap at the growing MT end. Here we use cryo-EM and TIRF microscopy of GTP hydrolysis-deficient MTs assembled from mutant recombinant human tubulin to investigate the structure of a GTP-bound MT lattice. We find that the GTP-MT lattice of two mutants in which the catalytically active glutamate in -tubulin was substituted by inactive amino acids (E254A and E254N) is remarkably plastic. Undecorated E254A and E254N MTs with 13 protofilaments both have an expanded lattice, but display opposite protofilament twists, making these lattices distinct from the compacted lattice of wildtype GDP-MTs. End binding proteins of the EB family have the ability to compact both mutant GTP-lattices and to stabilize a negative twist, suggesting that they promote this transition also in the GTP cap of wildtype MTs, thereby contributing to the maturation of the MT structure. We also find that the MT seam appears to be stabilized in mutant GTP-MTs and destabilized in GDP-MTs, supporting the proposal that the seam plays an important role in MT stability. Together, these first high-resolution structures of truly GTP-bound MTs add mechanistic insight to our understanding of MT dynamic instability.

Significance StatementMicrotubules (MTs) are non-equilibrium polymers that switch between states of growth and shrinkage. This property is critical for their function and is a consequence of GTP hydrolysis in the MT. The structure of the stable GTP part of the MT (the GTP cap) has previously been inferred from MTs polymerized with non-hydrolyzable GTP analogs. Here, we report the first high-resolution structures of MTs truly containing GTP, polymerized from mutated, hydrolysis-deficient tubulins. We find that GTP-MTs have an "expanded lattice" and a "closed seam", structural characteristics possibly responsible for stabilizing the GTP cap. These results provide new insight into the structural transitions at growing MT ends, furthering our understanding of the bistable nature of MTs.
]]></description>
<dc:creator>LaFrance, B. J.</dc:creator>
<dc:creator>Roostalu, J.</dc:creator>
<dc:creator>Henkin, G.</dc:creator>
<dc:creator>Greber, B. J.</dc:creator>
<dc:creator>Zhang, R.</dc:creator>
<dc:creator>Normanno, D.</dc:creator>
<dc:creator>McCollum, C.</dc:creator>
<dc:creator>Surrey, T.</dc:creator>
<dc:creator>Nogales, E.</dc:creator>
<dc:date>2021-08-13</dc:date>
<dc:identifier>doi:10.1101/2021.08.13.456308</dc:identifier>
<dc:title><![CDATA[Structural transitions in the GTP cap visualized by cryo-EM of catalytically inactive microtubules]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.08.13.456237v1?rss=1">
<title>
<![CDATA[
Autophagy coordinates chondrocyte development and early joint formation in zebrafish 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.08.13.456237v1?rss=1"
</link>
<description><![CDATA[
Autophagy is a catabolic process responsible for the removal of waste and damaged cellular components by lysosomal degradation. It plays a key role in fundamental cell processes, including ER stress mitigation, control of cell metabolism, and cell differentiation and proliferation, all of which are essential for cartilage cell (chondrocyte) development and survival, and for the formation of cartilage. Correspondingly, autophagy dysregulation has been implicated in several skeletal disorders such as osteoarthritis and osteoporosis. To test the requirement for autophagy during skeletal development in zebrafish, we generated an atg13 CRISPR knockout zebrafish line. This line showed a complete loss of atg13 expression, and restricted autophagic activity in vivo. In the absence of autophagy, chondrocyte maturation was accelerated, with chondrocytes exhibiting signs of premature hypertrophy. Focussing on the jaw element, autophagy disruption affected joint articulation causing restricted mouth opening. This gross behavioural phenotype corresponded with a failure to thrive, and death in homozygote atg13 nulls within 17 days. Taken together, our results are consistent with autophagy contributing to the timely regulation of chondrocyte maturation and for extracellular matrix formation.
]]></description>
<dc:creator>Moss, J. J.</dc:creator>
<dc:creator>Wirth, M.</dc:creator>
<dc:creator>Tooze, S. A.</dc:creator>
<dc:creator>Lane, J. D.</dc:creator>
<dc:creator>Hammond, C. L.</dc:creator>
<dc:date>2021-08-13</dc:date>
<dc:identifier>doi:10.1101/2021.08.13.456237</dc:identifier>
<dc:title><![CDATA[Autophagy coordinates chondrocyte development and early joint formation in zebrafish]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.08.16.456464v1?rss=1">
<title>
<![CDATA[
RSC-mediated nucleosome positioning and GRFs form barriers in promoters to limit divergent noncoding transcription 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.08.16.456464v1?rss=1"
</link>
<description><![CDATA[
The directionality of gene promoters - the ratio of protein-coding over divergent noncoding transcription - is highly variable and regulated. How promoter directionality is controlled remains poorly understood. Here, we show that the chromatin remodelling complex RSC and general regulatory factors (GRFs) dictate promoter directionality by attenuating divergent transcription. At gene promoters that are highly directional, depletion of RSC leads to a relative increase in divergent noncoding transcription and thus a decrease in promoter directionality. We find that RSC facilitates nucleosome positioning upstream in promoters at the sites of divergent transcription. These highly directional promoters are also enriched for the binding of GRFs such as Reb1 and Abf1. Ectopic targeting of divergent transcription initiation sites with GRFs or the dCas9 protein suppresses divergent transcription. Our data suggest that RSC and GRFs play a pervasive role in limiting divergent transcription. We propose that any DNA binding factor, when stably associated with cryptic transcription start sites, form barriers for repressing divergent transcription. Our study provides an explanation as to why certain promoters are more directional than others.
]]></description>
<dc:creator>Wu, A.</dc:creator>
<dc:creator>Vivori, C.</dc:creator>
<dc:creator>Patel, H.</dc:creator>
<dc:creator>Sideri, T.</dc:creator>
<dc:creator>van Werven, F.</dc:creator>
<dc:date>2021-08-16</dc:date>
<dc:identifier>doi:10.1101/2021.08.16.456464</dc:identifier>
<dc:title><![CDATA[RSC-mediated nucleosome positioning and GRFs form barriers in promoters to limit divergent noncoding transcription]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.08.18.456785v1?rss=1">
<title>
<![CDATA[
Direct reprogramming of human embryonic to trophoblast stem cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.08.18.456785v1?rss=1"
</link>
<description><![CDATA[
During the first week of development, human embryos form a blastocyst comprised of an inner cell mass and trophectoderm (TE) cells, the latter of which are progenitors of placental trophoblast. Here we investigated the expression of transcripts in the human TE from early to late blastocyst stages. We identified enrichment of transcription factors GATA2, GATA3, TFAP2C and KLF5 and characterised their protein expression dynamics across TE development. By inducible overexpression and mRNA transfection we determined that these factors, together with MYC, are sufficient to establish induced trophoblast stem cells (iTSCs) from primed human embryonic stem cells. These iTSCs self-renew and recapitulate morphological characteristics, gene expression profiles, and directed differentiation potential similar to existing human TSCs. Systematic omission of each, or combinations of factors, revealed the critical importance of GATA2 and GATA3 for iTSC transdifferentiation. Altogether, these findings provide insights into the transcription factor network that may be operational in the human TE and broaden the methods for establishing cellular models of early human placental progenitor cells, which may be useful in the future to model placental-associated diseases.

Summary statementTranscriptional analysis of human blastocysts reveals transcription factors sufficient to derive induced trophoblast stem cells from primed human embryonic stem cells.
]]></description>
<dc:creator>Fogarty, N. M.</dc:creator>
<dc:creator>Abdelbaki, A.</dc:creator>
<dc:creator>McCarthy, A.</dc:creator>
<dc:creator>Devito, L.</dc:creator>
<dc:creator>Chen, A. E.</dc:creator>
<dc:creator>Munusamy, P.</dc:creator>
<dc:creator>Blakeley, P.</dc:creator>
<dc:creator>Elder, K.</dc:creator>
<dc:creator>Snell, P.</dc:creator>
<dc:creator>Christie, L.</dc:creator>
<dc:creator>Serhal, P.</dc:creator>
<dc:creator>Odia, R. A.</dc:creator>
<dc:creator>Sangrithi, M.</dc:creator>
<dc:creator>Niakan, K. K.</dc:creator>
<dc:date>2021-08-18</dc:date>
<dc:identifier>doi:10.1101/2021.08.18.456785</dc:identifier>
<dc:title><![CDATA[Direct reprogramming of human embryonic to trophoblast stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.08.24.457560v1?rss=1">
<title>
<![CDATA[
Toxoplasma-proximal and distal control by GBPs in human macrophages 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.08.24.457560v1?rss=1"
</link>
<description><![CDATA[
Human guanylate-binding proteins (GBPs) are key players of interferon-gamma (IFN{gamma})-induced cell intrinsic defense mechanisms targeting intracellular pathogens. In this study we combine the well-established Toxoplasma gondii infection model with three in vitro macrophage culture systems to delineate the contribution of individual GBP family members to control this apicomplexan parasite. Use of high-throughput imaging assays and genome engineering allowed us to define a role for GBP1, 2 and 5 in parasite infection control. While GBP1 performs a pathogen-proximal, parasiticidal and growth-restricting function through accumulation at the parasitophorous vacuole of intracellular Toxoplasma, GBP2 and 5 perform a pathogen-distal, growth-restricting role. We further find that mutants of the GTPase or isoprenylation site of GBP1/2/5 affect their normal function in Toxoplasma control by leading to mis-localization of the proteins.
]]></description>
<dc:creator>Fisch, D.</dc:creator>
<dc:creator>Clough, B.</dc:creator>
<dc:creator>Khan, R. T.</dc:creator>
<dc:creator>Healy, L.</dc:creator>
<dc:creator>Frickel, E.</dc:creator>
<dc:date>2021-08-24</dc:date>
<dc:identifier>doi:10.1101/2021.08.24.457560</dc:identifier>
<dc:title><![CDATA[Toxoplasma-proximal and distal control by GBPs in human macrophages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.08.27.457890v1?rss=1">
<title>
<![CDATA[
An improved iCLIP protocol 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.08.27.457890v1?rss=1"
</link>
<description><![CDATA[
Crosslinking and Immunoprecipitation (CLIP) is a powerful technique to obtain transcriptome-wide maps of in vivo protein-RNA interactions, which are important to understand the post-transcriptional mechanisms mediated by RNA binding proteins (RBPs). Many variant CLIP protocols have been developed to improve the efficiency and convenience of cDNA library preparation. Here we describe an improved individual nucleotide resolution CLIP protocol (iiCLIP), which can be completed within 4 days from UV crosslinking to libraries for sequencing. For benchmarking, we directly compared PTBP1 iiCLIP libraries with the iCLIP2 protocol produced under standardised conditions, and with public eCLIP and iCLIP PTBP1 data. We visualised enriched motifs surrounding the identified crosslink positions and RNA maps of these crosslinks around the alternative exons regulated by PTBP1. Notably, motif enrichment was higher in iiCLIP and iCLIP2 in comparison to public eCLIP and iCLIP, and we show how this impacts the specificity of RNA maps. In conclusion, iiCLIP is technically convenient and efficient, and enables production of highly specific datasets for identifying RBP binding sites.
]]></description>
<dc:creator>Lee, F. C. Y.</dc:creator>
<dc:creator>Chakrabarti, A. M.</dc:creator>
<dc:creator>Hänel, H.</dc:creator>
<dc:creator>Monzon-Casanova, E.</dc:creator>
<dc:creator>Hallegger, M.</dc:creator>
<dc:creator>Militti, C.</dc:creator>
<dc:creator>Capraro, F.</dc:creator>
<dc:creator>Sadee, C.</dc:creator>
<dc:creator>Toolan-Kerr, P.</dc:creator>
<dc:creator>Wilkins, O.</dc:creator>
<dc:creator>Turner, M.</dc:creator>
<dc:creator>König, J.</dc:creator>
<dc:creator>Sibley, C.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:date>2021-08-27</dc:date>
<dc:identifier>doi:10.1101/2021.08.27.457890</dc:identifier>
<dc:title><![CDATA[An improved iCLIP protocol]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.08.31.458177v1?rss=1">
<title>
<![CDATA[
Comparative assessment of genes driving cancer and somatic evolution. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.08.31.458177v1?rss=1"
</link>
<description><![CDATA[
Genetic alterations of somatic cells can drive nonmalignant clone formation and promote cancer initiation. However, the link between these processes remains unclear hampering our understanding of tissue homeostasis and cancer development. Here we collect a literature-based repertoire of 3355 well-known or predicted drivers of cancer and noncancer somatic evolution in 122 cancer types and 12 noncancer tissues. Mapping the alterations of these genes in 7953 pancancer samples reveals that, despite the large size, the known compendium of drivers is still incomplete and biased towards frequently occurring coding mutations. High overlap exists between drivers of cancer and noncancer somatic evolution, although significant differences emerge in their recurrence. We confirm and expand the unique properties of drivers and identify a core of evolutionarily conserved and essential genes whose germline variation is strongly counter-selected. Somatic alteration in even one of these genes is sufficient to drive clonal expansion but not malignant transformation. Our study offers a comprehensive overview of our current understanding of the genetic events initiating clone expansion and cancer revealing significant gaps and biases that still need to be addressed. The compendium of cancer and noncancer somatic drivers, their literature support and properties are accessible at http://www.network-cancer-genes.org/.
]]></description>
<dc:creator>Dressler, L.</dc:creator>
<dc:creator>Bortolomeazzi, M.</dc:creator>
<dc:creator>Keddar, M. R.</dc:creator>
<dc:creator>Misetic, H.</dc:creator>
<dc:creator>Sartini, G.</dc:creator>
<dc:creator>Acha-Sagredo, A.</dc:creator>
<dc:creator>Montorsi, L.</dc:creator>
<dc:creator>Wijewardhane, N.</dc:creator>
<dc:creator>Repana, D.</dc:creator>
<dc:creator>Nulsen, J.</dc:creator>
<dc:creator>Goldman, J.</dc:creator>
<dc:creator>Pollit, M.</dc:creator>
<dc:creator>Davis, P.</dc:creator>
<dc:creator>Strange, A.</dc:creator>
<dc:creator>Ambrose, K.</dc:creator>
<dc:creator>Ciccarelli, F. D.</dc:creator>
<dc:date>2021-08-31</dc:date>
<dc:identifier>doi:10.1101/2021.08.31.458177</dc:identifier>
<dc:title><![CDATA[Comparative assessment of genes driving cancer and somatic evolution.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.08.30.458271v1?rss=1">
<title>
<![CDATA[
Axonal length determines distinct homeostatic phenotypes in human iPSC derived motor neurons on a bioengineered platform 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.08.30.458271v1?rss=1"
</link>
<description><![CDATA[
Stem cell-based experimental platforms for neuroscience can effectively model key mechanistic aspects of human development and disease. However, conventional culture systems often overlook the engineering constraints that cells face in vivo. This is particularly relevant for neurons covering long range connections such as spinal motor neurons (MNs). The axons of these neurons extend up to 1m in length and require a complex interplay of mechanisms to maintain cellular homeostasis. It follows that shorter axons in conventional cultures may not faithfully capture important aspects of their longer counterparts. Here we directly address this issue by establishing a bioengineered platform to assemble arrays of human axons ranging from micrometers to centimeters, permitting systematic investigation of the effects of length on human axonal biology for the first time. With this approach, we reveal a link between length and metabolism in human MNs in vitro, where axons above a "threshold" size induce specific molecular adaptations in cytoskeleton composition, functional properties, local translation and mitochondrial homeostasis. Our findings specifically demonstrate the existence of a length-dependent mechanism that switches homeostatic processes within human MNs in order to sustain long axons. Our findings have critical implications for in vitro modelling of several neurodegenerative disorders and reinforce the importance of modelling cell shape and biophysical constraints with fidelity and precision in vitro.
]]></description>
<dc:creator>Hagemann, C.</dc:creator>
<dc:creator>Moreno Gonzalez, C.</dc:creator>
<dc:creator>Guetta, L.</dc:creator>
<dc:creator>Tyzak, G.</dc:creator>
<dc:creator>Chiappini, C.</dc:creator>
<dc:creator>Legati, A.</dc:creator>
<dc:creator>Patani, R.</dc:creator>
<dc:creator>Serio, A.</dc:creator>
<dc:date>2021-09-01</dc:date>
<dc:identifier>doi:10.1101/2021.08.30.458271</dc:identifier>
<dc:title><![CDATA[Axonal length determines distinct homeostatic phenotypes in human iPSC derived motor neurons on a bioengineered platform]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.09.03.458852v1?rss=1">
<title>
<![CDATA[
Bringing TrackMate in the era of machine-learning and deep-learning. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.09.03.458852v1?rss=1"
</link>
<description><![CDATA[
TrackMate is an automated tracking software used to analyze bioimages and distributed as a Fiji plugin. Here we introduce a new version of TrackMate rewritten to improve performance and usability, and integrating several popular machine and deep learning algorithms to improve versatility. We illustrate how these new components can be used to efficiently track objects from brightfield and fluorescence microscopy images across a wide range of bio-imaging experiments.
]]></description>
<dc:creator>Ershov, D.</dc:creator>
<dc:creator>Phan, M.-S.</dc:creator>
<dc:creator>Pylvänäinen, J. W.</dc:creator>
<dc:creator>Rigaud, S. U.</dc:creator>
<dc:creator>Le Blanc, L.</dc:creator>
<dc:creator>Charles-Orszag, A.</dc:creator>
<dc:creator>Conway, J. R. W.</dc:creator>
<dc:creator>Laine, R. F.</dc:creator>
<dc:creator>Roy, N. H.</dc:creator>
<dc:creator>Bonazzi, D.</dc:creator>
<dc:creator>Dumenil, G.</dc:creator>
<dc:creator>Jacquemet, G.</dc:creator>
<dc:creator>Tinevez, J.-Y.</dc:creator>
<dc:date>2021-09-03</dc:date>
<dc:identifier>doi:10.1101/2021.09.03.458852</dc:identifier>
<dc:title><![CDATA[Bringing TrackMate in the era of machine-learning and deep-learning.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.09.09.459588v1?rss=1">
<title>
<![CDATA[
B cell-intrinsic requirement for WNK1 kinase in T cell-dependent antibody responses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.09.09.459588v1?rss=1"
</link>
<description><![CDATA[
Migration and adhesion play critical roles in B cells, regulating recirculation between lymphoid organs, migration within lymphoid tissue and interaction with CD4+ T cells. However, there is limited knowledge of how B cells integrate chemokine receptor and integrin signaling with B cell activation to generate efficient humoral responses. Here we show that the WNK1 kinase, a regulator of migration and adhesion, is essential in B cells for T-dependent antibody responses. We demonstrate that WNK1 transduces signals from the BCR, CXCR5 and CD40, and using intravital imaging we show that WNK1 regulates migration of naive and activated B cells, and their interactions with T cells. Unexpectedly, we show that WNK1 is required for BCR- and CD40-induced proliferation, acting through the OXSR1 and STK39 kinases, and for efficient B cell-T cell collaboration in vivo. Thus, WNK1 is critical for humoral immune responses, by regulating B cell migration, adhesion and T cell-dependent activation.

SummaryThe WNK1 kinase is essential in B cells for T-dependent antibody responses because it is activated by signaling from BCR, CXCR5 and CD40 and regulates B cell migration, adhesion, T-dependent activation, and differentiation into germinal center B cells and plasma cells.
]]></description>
<dc:creator>Hayward, D.</dc:creator>
<dc:creator>Vanes, L.</dc:creator>
<dc:creator>Wissmann, S.</dc:creator>
<dc:creator>Sivapatham, S.</dc:creator>
<dc:creator>Hartweger, H.</dc:creator>
<dc:creator>Biggs O'May, J.</dc:creator>
<dc:creator>De Boer, L.</dc:creator>
<dc:creator>Mitter, R.</dc:creator>
<dc:creator>Kochl, R.</dc:creator>
<dc:creator>Stein, J. V.</dc:creator>
<dc:creator>Tybulewicz, V. L.</dc:creator>
<dc:date>2021-09-09</dc:date>
<dc:identifier>doi:10.1101/2021.09.09.459588</dc:identifier>
<dc:title><![CDATA[B cell-intrinsic requirement for WNK1 kinase in T cell-dependent antibody responses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.09.07.459264v1?rss=1">
<title>
<![CDATA[
Fission stories: Using PomBase to understand Schizosaccharomyces pombe biology 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.09.07.459264v1?rss=1"
</link>
<description><![CDATA[
PomBase (www.pombase.org), the model organism database (MOD) for the fission yeast Schizosaccharomyces pombe, supports research within and beyond the S. pombe community by integrating and presenting genetic, molecular, and cell biological knowledge into intuitive displays and comprehensive data collections. With new content, novel query capabilities, and biologist-friendly data summaries and visualisation, PomBase also drives innovation in the MOD community.
]]></description>
<dc:creator>Harris, M. A.</dc:creator>
<dc:creator>Rutherford, K. M.</dc:creator>
<dc:creator>Hayles, J.</dc:creator>
<dc:creator>Lock, A.</dc:creator>
<dc:creator>Bähler, J.</dc:creator>
<dc:creator>Oliver, S. G.</dc:creator>
<dc:creator>Mata, J.</dc:creator>
<dc:creator>Wood, V.</dc:creator>
<dc:date>2021-09-10</dc:date>
<dc:identifier>doi:10.1101/2021.09.07.459264</dc:identifier>
<dc:title><![CDATA[Fission stories: Using PomBase to understand Schizosaccharomyces pombe biology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.09.10.459763v1?rss=1">
<title>
<![CDATA[
clipplotr - a comparative visualisation and analysis tool for CLIP data 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.09.10.459763v1?rss=1"
</link>
<description><![CDATA[
CLIP technologies are now widely used to study RNA-protein interactions and many datasets are now publicly available. An important first step in CLIP data exploration is the visual inspection and assessment of processed genomic data on selected genes or regions and performing comparisons: either across conditions within a particular project, or incorporating publicly available data. However, the output files produced by data processing pipelines or preprocessed files available to download from data repositories are often not suitable for direct comparison and usually need further processing. Furthermore, to derive biological insight it is usually necessary to visualise CLIP signal alongside other data such as annotations, or orthogonal functional genomic data (e.g. RNA-seq). We have developed a simple, but powerful, command-line tool: clipplotr, which facilitates these visual comparative and integrative analyses with normalisation and smoothing options for CLIP data and the ability to show these alongside reference annotation tracks and functional genomic data. These data can be supplied as input to clipplotr in a range of file formats, which will output a publication quality figure. It is written in R and can both run on a laptop computer independently, or be integrated into computational workflows on a high-performance cluster. Releases, source code and documentation are freely available at: https://github.com/ulelab/clipplotr.
]]></description>
<dc:creator>Chakrabarti, A. M.</dc:creator>
<dc:creator>Capitanchik, C.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Luscombe, N.</dc:creator>
<dc:date>2021-09-11</dc:date>
<dc:identifier>doi:10.1101/2021.09.10.459763</dc:identifier>
<dc:title><![CDATA[clipplotr - a comparative visualisation and analysis tool for CLIP data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.09.13.460021v1?rss=1">
<title>
<![CDATA[
Chromodomain protein regulates the expression of a subset of RIFINs in P. falciparum 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.09.13.460021v1?rss=1"
</link>
<description><![CDATA[
Plasmodium falciparum expresses clonally variant proteins on the surface of infected erythrocytes to evade the host immune system. The clonally variant multigenes include var, rifin, and stevor, which express EMP1, RIFIN, and STEVOR proteins, respectively. The rifins are the largest multigene family and are essentially involved in the RBC rosetting, the hallmark of severe malaria. The regulators that control the RIFINs expression in P. falciparum have not been reported so far. This study reports a chromodomain-containing protein (PfCDP) that binds to H3K9me3 modification on P. falciparum chromatin. The ChIP- sequencing analysis revealed that the PfCDP is majorly associated with clonally variant gene families, primarily rifins in P. falciparum. Conditional deletion of the chromodomain (CD) gene in P. falciparum leads to the up-regulation of a subset of virulence genes, including rifins, a few var, and stevor genes. Further, we show that Pf{Delta}CDP P. falciparum lines promote the RBC rosetting. This study provides evidence of an epigenetic regulator mediated control on a subset of RIFINs expression and RBC rosetting by P. falciparum.
]]></description>
<dc:creator>Sethumadhavan, D. V.</dc:creator>
<dc:creator>Tiburcio, M.</dc:creator>
<dc:creator>Kanyal, A.</dc:creator>
<dc:creator>Jabeena, C.</dc:creator>
<dc:creator>Govindaraju, G.</dc:creator>
<dc:creator>Karmodiya, K.</dc:creator>
<dc:creator>Rajavelu, A.</dc:creator>
<dc:date>2021-09-13</dc:date>
<dc:identifier>doi:10.1101/2021.09.13.460021</dc:identifier>
<dc:title><![CDATA[Chromodomain protein regulates the expression of a subset of RIFINs in P. falciparum]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.07.20.453060v1?rss=1">
<title>
<![CDATA[
BashTheBug: a crowd of volunteers reproducibly and accurately measure the minimum inhibitory concentrations of 13 antitubercular drugs from photographs of 96-well broth microdilution plates. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.07.20.453060v1?rss=1"
</link>
<description><![CDATA[
Tuberculosis is a respiratory disease that is treatable with antibiotics. An increasing prevalence of resistance means that to ensure a good treatment outcome it is desirable to test the susceptibility of each infection to different antibiotics. Conventionally this is done by culturing a clinical sample and then exposing aliquots to a panel of antibiotics, thereby determining the minimum inhibitory concentration (MIC) of each drug. Using 96-well broth micro dilution plates with each well containing a lyophilised pre-determined amount of an antibiotic is a convenient and cost-effective way to measure the MICs of several drugs at once for a clinical sample. Although accurate, this is an expensive and slow process that requires highly-skilled and experienced laboratory scientists. Here we show that, through the BashTheBug project hosted on the Zooniverse citizen science platform, a crowd of volunteers can reproducibly and accurately determine the MICs for 13 drugs and that simply taking the median or mode of 11-17 independent classifications is sufficient. There is therefore a potential role for crowds to support (but not supplant) the role of experts in antibiotic susceptibility testing.
]]></description>
<dc:creator>Fowler, P. W.</dc:creator>
<dc:creator>Wright, C.</dc:creator>
<dc:creator>Spiers-Bowers, H.</dc:creator>
<dc:creator>Zhu, T.</dc:creator>
<dc:creator>Baeten, E. M.</dc:creator>
<dc:creator>Hoosdally, S. W.</dc:creator>
<dc:creator>Gibertoni Cruz, A. L.</dc:creator>
<dc:creator>Roohi, A.</dc:creator>
<dc:creator>Kouchaki, S.</dc:creator>
<dc:creator>Walker, T. M.</dc:creator>
<dc:creator>Peto, T. E.</dc:creator>
<dc:creator>Miller, G.</dc:creator>
<dc:creator>Lintott, C.</dc:creator>
<dc:creator>Clifton, D.</dc:creator>
<dc:creator>Crook, D. W.</dc:creator>
<dc:creator>Walker, A. S.</dc:creator>
<dc:creator>The CRyPTIC Consortium,</dc:creator>
<dc:date>2021-07-21</dc:date>
<dc:identifier>doi:10.1101/2021.07.20.453060</dc:identifier>
<dc:title><![CDATA[BashTheBug: a crowd of volunteers reproducibly and accurately measure the minimum inhibitory concentrations of 13 antitubercular drugs from photographs of 96-well broth microdilution plates.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.09.08.459433v1?rss=1">
<title>
<![CDATA[
Comprehensive analysis of DNA replication timing in genetic diseases and gene knockouts identifies MCM10 as a novel regulator of the replication program 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.09.08.459433v1?rss=1"
</link>
<description><![CDATA[
Cellular proliferation depends on the accurate and timely replication of the genome. Several genetic diseases are caused by mutations in key DNA replication genes; however, it remains unclear whether these genes influence the normal program of DNA replication timing. Similarly, the factors that regulate DNA replication dynamics are poorly understood. To systematically identify trans-acting modulators of replication timing, we profiled replication in 184 cell lines from three cell types, encompassing 60 different gene knockouts or genetic diseases. Through a rigorous approach that considers the background variability of replication timing, we concluded that most samples displayed normal replication timing. However, mutations in two genes showed consistently abnormal replication timing. The first gene was RIF1, a known modulator of replication timing. The second was MCM10, a highly conserved member of the pre-replication complex. MCM10 mutant cells demonstrated replication timing variability comprising 46% of the genome and at different locations than RIF1 knockouts. Replication timing alterations in MCM10-mutant cells was predominantly comprised of replication initiation defects. Taken together, this study demonstrates the remarkable robustness of the human replication timing program and reveals MCM10 as a novel modulator of DNA replication timing.
]]></description>
<dc:creator>Caballero, M.</dc:creator>
<dc:creator>Ge, T.</dc:creator>
<dc:creator>Rebelo, A. R.</dc:creator>
<dc:creator>Seo, S.</dc:creator>
<dc:creator>Kim, S.</dc:creator>
<dc:creator>Brooks, K.</dc:creator>
<dc:creator>Zuccaro, M. L.</dc:creator>
<dc:creator>Kanagaraj, R.</dc:creator>
<dc:creator>Vershkov, D.</dc:creator>
<dc:creator>Kim, D.</dc:creator>
<dc:creator>Smogorzewska, A.</dc:creator>
<dc:creator>Smolka, M. B.</dc:creator>
<dc:creator>Benvenisty, N.</dc:creator>
<dc:creator>West, S. C.</dc:creator>
<dc:creator>Egli, D. C.</dc:creator>
<dc:creator>Mace, E. M.</dc:creator>
<dc:creator>Koren, A.</dc:creator>
<dc:date>2021-09-09</dc:date>
<dc:identifier>doi:10.1101/2021.09.08.459433</dc:identifier>
<dc:title><![CDATA[Comprehensive analysis of DNA replication timing in genetic diseases and gene knockouts identifies MCM10 as a novel regulator of the replication program]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.09.14.460335v1?rss=1">
<title>
<![CDATA[
Cdc6 is sequentially regulated by PP2A-Cdc55, Cdc14 and Sic1 for origin licensing in S. cerevisiae 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.09.14.460335v1?rss=1"
</link>
<description><![CDATA[
Cdc6, a subunit of the pre-replicative complex, contains multiple regulatory Cdk1 consensus sites, SP or TP motifs. In S. cerevisiae, Cdk1 phosphorylates Cdc6-T7 to recruit Cks1, the Cdk1 phospho-adaptor in S-phase, for subsequent multisite phosphorylation and protein degradation. Cdc6 accumulates in mitosis and is tightly bound by Clb2 through N-terminal phosphorylation in order to prevent premature origin licensing and degradation. It has been extensively studied how Cdc6 phosphorylation is regulated by the Cyclin-Cdk1 complex. However, a detailed mechanism on how Cdc6 phosphorylation is reversed by phosphatases has not been elucidated. Here, we show that PP2ACdc55 dephosphorylates Cdc6 N-terminal sites to release Clb2. Cdc14 dephosphorylates the C-terminal phospho-degron, leading to Cdc6 stabilization in mitosis. In addition, the Cdk1 inhibitor, Sic1, releases Clb2{middle dot}Cdk1{middle dot}Cks1 from Cdc6 to load Mcm2-7 on the chromatin upon mitotic exit. Thus, pre-RC assembly and origin licensing is promoted by the attenuation of distinct CDK-dependent Cdc6 inhibitory mechanisms.
]]></description>
<dc:creator>Philip, J.</dc:creator>
<dc:creator>Ord, M.</dc:creator>
<dc:creator>Silva, A.</dc:creator>
<dc:creator>Singh, S.</dc:creator>
<dc:creator>Diffley, J. F. X.</dc:creator>
<dc:creator>Remus, D.</dc:creator>
<dc:creator>Loog, M.</dc:creator>
<dc:creator>Ikui, A. E.</dc:creator>
<dc:date>2021-09-15</dc:date>
<dc:identifier>doi:10.1101/2021.09.14.460335</dc:identifier>
<dc:title><![CDATA[Cdc6 is sequentially regulated by PP2A-Cdc55, Cdc14 and Sic1 for origin licensing in S. cerevisiae]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.09.14.460272v1?rss=1">
<title>
<![CDATA[
Genome-wide association studies of global Mycobacterium tuberculosis resistance to thirteen antimicrobials in 10,228 genomes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.09.14.460272v1?rss=1"
</link>
<description><![CDATA[
The emergence of drug resistant tuberculosis is a major global public health concern that threatens the ability to control the disease. Whole genome sequencing as a tool to rapidly diagnose resistant infections can transform patient treatment and clinical practice. While resistance mechanisms are well understood for some drugs, there are likely many mechanisms yet to be uncovered, particularly for new and repurposed drugs. We sequenced 10,228 Mycobacterium tuberculosis (MTB) isolates worldwide and determined the minimum inhibitory concentration (MIC) on a grid of twofold concentration dilutions for 13 antimicrobials using quantitative microtiter plate assays. We performed oligopeptide- and oligonucleotide-based genome-wide association studies using linear mixed models to discover resistance-conferring mechanisms not currently catalogued. Use of MIC over binary resistance phenotypes increased heritability for the new and repurposed drugs by 26-37%, increasing our ability to detect novel associations. For all drugs, we discovered uncatalogued variants associated with MIC, including in the Rv1218c promoter binding site of the transcriptional repressor Rv1219c (isoniazid), upstream of the vapBC20 operon that cleaves 23S rRNA (linezolid) and in the region encoding an -helix lining the active site of Cyp142 (clofazimine, all p<10-7.7). We observed that artefactual signals of cross resistance could be unravelled based on the relative effect size on MIC. Our study demonstrates the ability of very large-scale studies to substantially improve our knowledge of genetic variants associated with antimicrobial resistance in M. tuberculosis.
]]></description>
<dc:creator>Earle, S. G.</dc:creator>
<dc:creator>Wilson, D. J.</dc:creator>
<dc:creator>The CRyPTIC Consortium,</dc:creator>
<dc:date>2021-09-16</dc:date>
<dc:identifier>doi:10.1101/2021.09.14.460272</dc:identifier>
<dc:title><![CDATA[Genome-wide association studies of global Mycobacterium tuberculosis resistance to thirteen antimicrobials in 10,228 genomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.09.14.460274v1?rss=1">
<title>
<![CDATA[
A data compendium of Mycobacterium tuberculosis antibiotic resistance 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.09.14.460274v1?rss=1"
</link>
<description><![CDATA[
The Comprehensive Resistance Prediction for Tuberculosis: an International Consortium (CRyPTIC) presents here a compendium of 15,211 Mycobacterium tuberculosis global clinical isolates, all of which have undergone whole genome sequencing (WGS) and have had their minimum inhibitory concentrations to 13 antitubercular drugs measured in a single assay. It is the largest matched phenotypic and genotypic dataset for M. tuberculosis to date. Here, we provide a summary detailing the breadth of data collected, along with a description of how the isolates were collected and uniformly processed in CRyPTIC partner laboratories across 23 countries. The compendium contains 6,814 isolates resistant to at least one drug, including 2,129 samples that fully satisfy the clinical definitions of rifampicin resistant (RR), multi-drug resistant (MDR), pre-extensively drug resistant (pre-XDR) or extensively drug resistant (XDR). Accurate prediction of resistance status (sensitive/resistant) to eight antitubercular drugs by using a genetic mutation catalogue is presented along with the presence of suspected resistance-conferring mutations for isolates resistant to the newly introduced drugs bedaquiline, clofazimine, delamanid and linezolid. Finally, a case study of rifampicin mono-resistance demonstrates how this compendium could be used to advance our genetic understanding of rare resistance phenotypes. The compendium is fully open-source and it is hoped that the dataset will facilitate and inspire future research for years to come.
]]></description>
<dc:creator>Brankin, A.</dc:creator>
<dc:creator>Malone, K. M.</dc:creator>
<dc:creator>The CRyPTIC Consortium,</dc:creator>
<dc:date>2021-09-15</dc:date>
<dc:identifier>doi:10.1101/2021.09.14.460274</dc:identifier>
<dc:title><![CDATA[A data compendium of Mycobacterium tuberculosis antibiotic resistance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.09.14.458035v1?rss=1">
<title>
<![CDATA[
A generalisable approach to drug susceptibility prediction for M. Tuberculosis using machine learning and whole-genome sequencing 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.09.14.458035v1?rss=1"
</link>
<description><![CDATA[
There remains a clinical need for better approaches to rapid drug susceptibility testing in view of the increasing burden of multidrug resistant tuberculosis. Binary susceptibility phenotypes only capture changes in minimum inhibitory concentration when these cross the critical concentration, even though other changes may be clinically relevant. We developed a machine learning system to predict minimum inhibitory concentration from unassembled whole-genome sequencing data for 13 anti-tuberculosis drugs. We trained, validated and tested the system on 10,859 isolates from the CRyPTIC dataset. Essential agreement rates (predicted MIC within one doubling dilution of observed MIC) were above 92% for first-line drugs, 91% for fluoroquinolones and aminoglycosides, and 90% for new and repurposed drugs, albeit with a significant drop in performance for the very few phenotypically resistant isolates in the latter group. To further validate the model in the absence of external MIC datasets, we predicted MIC and converted values to binary for an external set of 15,239 isolates with binary phenotypes, and compare their performance against a previously validated mutation catalogue, the expected performance of existing molecular assays, and World Health Organization Target Product Profiles. The sensitivity of the model on the external dataset was greater than 90% for all drugs except ethionamide, clofazimine and linezolid. Specificity was greater than 95% for all drugs except ethambutol, ethionamide, bedaquiline, delamanid and clofazimine. The proposed system can provide quantitative susceptibility phenotyping to help guide antimicrobial therapy, although further data collection and validation are required before machine learning can be used clinically for all drugs.
]]></description>
<dc:creator>The CRyPTIC consortium,</dc:creator>
<dc:creator>Lachapelle, A. S.</dc:creator>
<dc:date>2021-09-16</dc:date>
<dc:identifier>doi:10.1101/2021.09.14.458035</dc:identifier>
<dc:title><![CDATA[A generalisable approach to drug susceptibility prediction for M. Tuberculosis using machine learning and whole-genome sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.09.19.460961v1?rss=1">
<title>
<![CDATA[
The master energy homeostasis regulator PGC-1alpha couples transcriptional co-activation and mRNA nuclear export 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.09.19.460961v1?rss=1"
</link>
<description><![CDATA[
PGC-1 plays a central role in maintaining the mitochondrial and energy metabolism homeostasis, linking external stimuli to the transcriptional co-activation of genes involved in adaptive and age-related pathways. The carboxyl-terminus encodes a serine/arginine-rich (RS) region and a putative RNA recognition motif, however potential RNA-processing role(s) have remained elusive for the past 20 years. Here, we show that the RS domain of human PGC-1 directly interacts with RNA and the nuclear RNA export factor NXF1. Inducible depletion of endogenous PGC-1 and expression of RNAi-resistant RS-deleted PGC-1 further demonstrate that the RNA-binding activity is required for nuclear export of co-activated transcripts and mitochondrial homeostasis. Moreover, a quantitative proteomics approach confirmed PGC-1-dependent RNA transport and mitochondrial-related functions, identifying also novel mRNA nuclear export targets in age-related telomere maintenance. Discovering a novel function for a major cellular homeostasis regulator provides new directions to further elucidate the roles of PGC-1 in gene expression, metabolic disorders, ageing and neurodegenerative diseases.
]]></description>
<dc:creator>Mihaylov, S. R.</dc:creator>
<dc:creator>Castelli, L. M.</dc:creator>
<dc:creator>Lin, Y.-H.</dc:creator>
<dc:creator>Gul, A.</dc:creator>
<dc:creator>Soni, N.</dc:creator>
<dc:creator>Hastings, C.</dc:creator>
<dc:creator>Flynn, H. R.</dc:creator>
<dc:creator>Dickman, M. J.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Bandmann, O.</dc:creator>
<dc:creator>Mortiboys, H.</dc:creator>
<dc:creator>Ultanir, S. K.</dc:creator>
<dc:creator>Hautbergue, G. M.</dc:creator>
<dc:date>2021-09-19</dc:date>
<dc:identifier>doi:10.1101/2021.09.19.460961</dc:identifier>
<dc:title><![CDATA[The master energy homeostasis regulator PGC-1alpha couples transcriptional co-activation and mRNA nuclear export]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.09.20.461005v1?rss=1">
<title>
<![CDATA[
RNA polymerase II dynamics and mRNA stability feedback determine mRNA scaling with cell size 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.09.20.461005v1?rss=1"
</link>
<description><![CDATA[
A fundamental feature of cellular growth is that protein and RNA amounts scale with cell size so that concentrations remain constant. A key component to this is that global transcription rates increase in larger cells, but the underlying mechanism has remained unknown. Here, we identify RNAPII as the major limiting factor increasing transcription with cell size in budding yeast as transcription is highly sensitive to the dosage of RNAPII but not to other components of the general transcriptional machinery. Our experiments support a dynamic equilibrium model where global transcription at a given size is set by the mass-action recruitment kinetics of unengaged nucleoplasmic RNAPII, and DNA content. This drives a sub-linear increase in transcription with size, which is precisely compensated for by a decrease in mRNA decay rates as cells enlarge. Thus, limiting RNAPII and feedback on mRNA stability work in concert to ensure mRNA concentration homeostasis in growing cells.
]]></description>
<dc:creator>Swaffer, M. P.</dc:creator>
<dc:creator>Marinov, G. K.</dc:creator>
<dc:creator>Zheng, H.</dc:creator>
<dc:creator>Jones, A. W.</dc:creator>
<dc:creator>Kundaje, A.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Greenleaf, W. J.</dc:creator>
<dc:creator>Reyes-Lamothe, R.</dc:creator>
<dc:creator>Skotheim, J. M.</dc:creator>
<dc:date>2021-09-20</dc:date>
<dc:identifier>doi:10.1101/2021.09.20.461005</dc:identifier>
<dc:title><![CDATA[RNA polymerase II dynamics and mRNA stability feedback determine mRNA scaling with cell size]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.09.14.460353v1?rss=1">
<title>
<![CDATA[
Quantitative measurement of antibiotic resistance in Mycobacterium tuberculosis reveals genetic determinants of resistance and susceptibility in a target gene approach 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.09.14.460353v1?rss=1"
</link>
<description><![CDATA[
The World Health Organization goal of universal drug susceptibility testing for patients with tuberculosis is most likely to be achieved through molecular diagnostics; however, to date these have focused largely on first-line drugs, and always on predicting binary susceptibilities. Here, we used whole genome sequencing and a quantitative microtiter plate assay to relate genomic mutations to minimum inhibitory concentration in 15,211 Mycobacterium tuberculosis patient isolates from 27 countries across five continents.

This work identifies 449 unique MIC-elevating genetic determinants across thirteen drugs, as well as 91 mutations resulting in hypersensitivity for eleven drugs. Our results provide a guide for further implementation of personalized medicine for the treatment of tuberculosis using genetics-based diagnostics and can serve as a training set for novel approaches to predict drug resistance.
]]></description>
<dc:creator>CRyPTIC Consortium,</dc:creator>
<dc:creator>Carter, J. J.</dc:creator>
<dc:date>2021-09-16</dc:date>
<dc:identifier>doi:10.1101/2021.09.14.460353</dc:identifier>
<dc:title><![CDATA[Quantitative measurement of antibiotic resistance in Mycobacterium tuberculosis reveals genetic determinants of resistance and susceptibility in a target gene approach]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.09.23.461587v1?rss=1">
<title>
<![CDATA[
JaponicusDB: Rapid deployment of a model organism database for an emerging model species 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.09.23.461587v1?rss=1"
</link>
<description><![CDATA[
The fission yeast Schizosaccharomyces japonicus has recently emerged as a powerful system for studying the evolution of essential cellular processes, drawing on similarities as well as key differences between S. japonicus and the related, well-established model Schizosaccharomyces pombe. We have deployed the open-source, modular code and tools originally developed for PomBase, the S. pombe model organism database (MOD), to create JaponicusDB (www.japonicusdb.org), a new MOD dedicated to S. japonicus. By providing a central resource with ready access to a growing body of experimental data, ontology-based curation, seamless browsing and querying, and the ability to integrate new data with existing knowledge, JaponicusDB supports fission yeast biologists to a far greater extent than any other source of S. japonicus data. JaponicusDB thus enables S. japonicus researchers to realise the full potential of studying a newly emerging model species, and illustrates the widely applicable power and utility of harnessing reusable PomBase code to build a comprehensive, community-maintainable repository of species-relevant knowledge.
]]></description>
<dc:creator>Rutherford, K. M.</dc:creator>
<dc:creator>Harris, M. A.</dc:creator>
<dc:creator>Oliferenko, S.</dc:creator>
<dc:creator>Wood, V.</dc:creator>
<dc:date>2021-09-24</dc:date>
<dc:identifier>doi:10.1101/2021.09.23.461587</dc:identifier>
<dc:title><![CDATA[JaponicusDB: Rapid deployment of a model organism database for an emerging model species]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.09.15.460475v1?rss=1">
<title>
<![CDATA[
Minos: variant adjudication and joint genotyping of cohorts of bacterial genomes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.09.15.460475v1?rss=1"
</link>
<description><![CDATA[
Short-read variant calling for bacterial genomics is a mature field, and there are many widely-used software tools. Different underlying approaches (eg pileup, local or global assembly, paired-read use, haplotype use) lend each tool different strengths, especially when considering non-SNP (single nucleotide polymorphism) variation or potentially distant reference genomes. It would therefore be valuable to be able to integrate the results from multiple variant callers, using a robust statistical approach to "adjudicate" at loci where there is disagreement between callers. To this end, we present a tool, Minos, for variant adjudication by mapping reads to a genome graph of variant calls. Minos allows users to combine output from multiple variant callers without loss of precision. Minos also addresses a second problem of joint genotyping SNPs and indels in bacterial cohorts, which can also be framed as an adjudication problem.

We benchmark on 62 samples from 3 species (Mycobacterium tuberculosis, Staphylococcus aureus, Klebsiella pneumoniae) and an outbreak of 385 M. tuberculosis samples. Finally, we joint genotype a large M. tuberculosis cohort (N{approx}15k) for which the rifampicin phenotype is known. We build a map of non-synonymous variants in the RRDR (rifampicin resistance determining region) of the rpoB gene and extend current knowledge relating RRDR SNPs to heterogeneity in rifampicin resistance levels. We replicate this finding in a second M. tuberculosis cohort (N{approx}13k).

Minos is released under the MIT license, available at https://github.com/iqbal-lab-org/minos.
]]></description>
<dc:creator>Hunt, M.</dc:creator>
<dc:creator>Letcher, B.</dc:creator>
<dc:creator>Malone, K. M.</dc:creator>
<dc:creator>Nguyen, G.</dc:creator>
<dc:creator>Hall, M. B.</dc:creator>
<dc:creator>Colquhoun, R. M.</dc:creator>
<dc:creator>Lima, L.</dc:creator>
<dc:creator>Schatz, M.</dc:creator>
<dc:creator>Ramakrishnan, S.</dc:creator>
<dc:creator>The CRyPTIC Consortium,</dc:creator>
<dc:creator>Iqbal, Z.</dc:creator>
<dc:date>2021-09-15</dc:date>
<dc:identifier>doi:10.1101/2021.09.15.460475</dc:identifier>
<dc:title><![CDATA[Minos: variant adjudication and joint genotyping of cohorts of bacterial genomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.10.07.463509v1?rss=1">
<title>
<![CDATA[
The Relative Binding Position of Nck and Grb2 Adaptors Dramatically Impacts Actin-Based Motility of Vaccinia Virus 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.10.07.463509v1?rss=1"
</link>
<description><![CDATA[
Phosphotyrosine (pTyr) motifs in unstructured polypeptides orchestrate important cellular processes by engaging SH2-containing adaptors to nucleate complex signalling networks. The concept of phase separation has recently changed our appreciation of such multivalent networks, however, the role of pTyr motif positioning in their function remains to be explored. We have now explored this parameter in the assembly and operation of the signalling cascade driving actin-based motility and spread of Vaccinia virus. This network involves two pTyr motifs in the viral protein A36 that recruit the adaptors Nck and Grb2 upstream of N-WASP and Arp2/3-mediated actin polymerization. We generated synthetic networks on Vaccinia by manipulating pTyr motifs in A36 and the unrelated p14 from Orthoreovirus. In contrast to predictions, we find that only specific spatial arrangements of Grb2 and Nck binding sites result in robust N-WASP recruitment, Arp2/3 driven actin polymerization and viral spread. Our results suggest that the relative position of pTyr adaptor binding sites is optimised for signal output. This finding may explain why the relative positions of pTyr motifs are usually conserved in proteins from widely different species. It also has important implications for regulation of physiological networks, including those that undergo phase transitions.
]]></description>
<dc:creator>Basant, A.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:date>2021-10-08</dc:date>
<dc:identifier>doi:10.1101/2021.10.07.463509</dc:identifier>
<dc:title><![CDATA[The Relative Binding Position of Nck and Grb2 Adaptors Dramatically Impacts Actin-Based Motility of Vaccinia Virus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.10.08.463636v1?rss=1">
<title>
<![CDATA[
Low dose pig anti-influenza virus monoclonal antibodies reduce lung pathology but do not prevent virus shedding 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.10.08.463636v1?rss=1"
</link>
<description><![CDATA[
We have established the pig, a large natural host animal for influenza, with many physiological similarities to humans, as a robust model for testing the therapeutic potential of monoclonal antibodies (mAbs). In this study we demonstrated that prophylactic intravenous administration of 15mg/kg of porcine mAb pb18, against the K160-163 site of the haemagglutinin, significantly reduced lung pathology and nasal virus shedding and eliminated virus from the lung of pigs following H1N1pdm09 challenge. When given at 1mg/kg, pb18 significantly reduced lung pathology and lung and BAL virus loads, but not nasal shedding. Similarly, when pb18 was given in combination with pb27, which recognised the K130 site, at 1mg/kg each, lung virus load and pathology were reduced, although without an apparent additive or synergistic effect. No evidence for mAb driven virus evolution was detected. These data indicate that intravenous administration of high doses was required to reduce nasal virus shedding, although this was inconsistent and seldom complete. In contrast the effect on lung pathology and lung virus load is consistent and is also seen at one log lower doses, strongly indicating that a lower dose might be sufficient to reduce severity of disease, but for prevention of transmission other measures would be needed.
]]></description>
<dc:creator>Paudyal, B.</dc:creator>
<dc:creator>McNee, A.</dc:creator>
<dc:creator>Rijal, P.</dc:creator>
<dc:creator>Carr, B. V.</dc:creator>
<dc:creator>Nunez, A.</dc:creator>
<dc:creator>McCauley, J. W.</dc:creator>
<dc:creator>Daniels, R. S.</dc:creator>
<dc:creator>Townsend, A. R.</dc:creator>
<dc:creator>Hammond, J. A.</dc:creator>
<dc:creator>Tchilian, E.</dc:creator>
<dc:date>2021-10-08</dc:date>
<dc:identifier>doi:10.1101/2021.10.08.463636</dc:identifier>
<dc:title><![CDATA[Low dose pig anti-influenza virus monoclonal antibodies reduce lung pathology but do not prevent virus shedding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.10.15.464407v1?rss=1">
<title>
<![CDATA[
Single-Shot Optical Projection tomography for high-speed volumetric imaging 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.10.15.464407v1?rss=1"
</link>
<description><![CDATA[
We present a single-shot adaptation of Optical Projection Tomography (OPT) for high-speed volumetric snapshot imaging of dynamic mesoscopic samples. Conventional OPT has been applied to in vivo imaging of animal models such as D. rerio but the sequential acquisition of projection images required for volumetric reconstruction typically requires samples to be immobilised during the acquisition of an OPT data set. We present a proof-of-principle system capable of single-shot imaging of a 1 mm diameter volume, demonstrating camera-limited rates of up to 62.5 volumes/second, which we have applied to 3D imaging of a freely-swimming zebrafish embryo. This is achieved by recording 8 projection views simultaneously on 4 low-cost CMOS cameras. With no stage required to rotate the sample, this single-shot OPT system can be implemented with a component cost of under {pound}5,000. The system design can be adapted to different sized fields of view and may be applied to a broad range of dynamic samples, including fluid dynamics.
]]></description>
<dc:creator>Darling, C. J.</dc:creator>
<dc:creator>Davis, S. P. X.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>French, P. M. W.</dc:creator>
<dc:creator>McGinty, J. A.</dc:creator>
<dc:date>2021-10-16</dc:date>
<dc:identifier>doi:10.1101/2021.10.15.464407</dc:identifier>
<dc:title><![CDATA[Single-Shot Optical Projection tomography for high-speed volumetric imaging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.10.18.464819v1?rss=1">
<title>
<![CDATA[
Therapeutic KRASG12C inhibition drives effective interferon-mediated anti-tumour immunity in immunogenic lung cancers 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.10.18.464819v1?rss=1"
</link>
<description><![CDATA[
Recently developed KRASG12C inhibitory drugs are beneficial to lung cancer patients harbouring KRASG12C mutations, but drug resistance frequently develops. Due to the immunosuppressive nature of the signaling network controlled by oncogenic KRAS, these drugs can indirectly affect anti-tumour immunity, providing a rationale for their combination with immune checkpoint blockade. In this study, we have characterised how KRASG12C inhibition reverses immune suppression driven by oncogenic KRAS in a number of pre-clinical lung cancer models with varying levels of immunogenicity. Mechanistically, KRASG12C inhibition upregulates interferon signaling via Myc inhibition, leading to reduced tumour infiltration by immunosuppressive cells, enhanced infiltration and activation of cytotoxic T cells, and increased antigen presentation. However, the combination of KRASG12C inhibitors with immune checkpoint blockade only provides synergistic benefit in the most immunogenic tumour model. KRASG12C inhibition fails to sensitize cold tumours to immunotherapy, with implications for the design of clinical trials combining KRASG12C inhibitors with anti-PD1 drugs.

One sentence summaryKRAS inhibition mobilizes anti-tumour immunity in immunogenic lung cancer models through derepressing interferon signaling via repression of Myc.
]]></description>
<dc:creator>Mugarza, E.</dc:creator>
<dc:creator>van Maldegem, F.</dc:creator>
<dc:creator>Boumelha, J.</dc:creator>
<dc:creator>Moore, C.</dc:creator>
<dc:creator>Rana, S.</dc:creator>
<dc:creator>Sopena, M. L.</dc:creator>
<dc:creator>East, P.</dc:creator>
<dc:creator>Ambler, R.</dc:creator>
<dc:creator>Anastasiou, P.</dc:creator>
<dc:creator>Clavijo, P. R.</dc:creator>
<dc:creator>Valand, K.</dc:creator>
<dc:creator>Cole, M.</dc:creator>
<dc:creator>Molina-Arcas, M.</dc:creator>
<dc:creator>Downward, J.</dc:creator>
<dc:date>2021-10-18</dc:date>
<dc:identifier>doi:10.1101/2021.10.18.464819</dc:identifier>
<dc:title><![CDATA[Therapeutic KRASG12C inhibition drives effective interferon-mediated anti-tumour immunity in immunogenic lung cancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.10.20.464849v1?rss=1">
<title>
<![CDATA[
Combinatorial tumor suppressor inactivation efficiently initiates lung adenocarcinoma with therapeutic vulnerabilities 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.10.20.464849v1?rss=1"
</link>
<description><![CDATA[
Lung cancer is the leading cause of cancer death worldwide, with lung adenocarcinoma being the most common subtype. Many oncogenes and tumor suppressor genes are altered in this cancer type and the discovery of oncogene mutations has led to the development of targeted therapies that have improved clinical outcomes. However, a large fraction of lung adenocarcinomas lacks mutations in known oncogenes, and the genesis and treatment of these oncogene-negative tumors remain enigmatic. Here, we perform iterative in vivo functional screens using quantitative autochthonous mouse model systems to uncover the genetic and biochemical changes that enable efficient lung tumor initiation in the absence of oncogene alterations. Through the generation of hundreds of diverse combinations of tumor suppressor alterations, we demonstrate that the inactivation of suppressors of the RAS and PI3K pathways drive the development of oncogene-negative lung adenocarcinoma. Human genomic data and histology identified RAS/MAPK and PI3K pathway activation as a common event in oncogene- negative human lung adenocarcinomas. We demonstrate that these Onc-negativeRAS/PI3K tumors and related cell lines are vulnerable to pharmacological inhibition of these signaling axes. These results transform our understanding of this prevalent yet understudied subtype of lung adenocarcinoma.
]]></description>
<dc:creator>Yousefi, M.</dc:creator>
<dc:creator>Boross, G.</dc:creator>
<dc:creator>Weiss, C.</dc:creator>
<dc:creator>Murray, C. W.</dc:creator>
<dc:creator>Hebert, J. D.</dc:creator>
<dc:creator>Cai, H.</dc:creator>
<dc:creator>Ashkin, E. L.</dc:creator>
<dc:creator>Karmakar, S.</dc:creator>
<dc:creator>Andrejka, L.</dc:creator>
<dc:creator>Chen, L.</dc:creator>
<dc:creator>Wang, M.</dc:creator>
<dc:creator>Tsai, M. K.</dc:creator>
<dc:creator>Lin, W.-Y.</dc:creator>
<dc:creator>Li, C.</dc:creator>
<dc:creator>Yakhchalian, P.</dc:creator>
<dc:creator>Colon, C. I.</dc:creator>
<dc:creator>Chew, S.-K.</dc:creator>
<dc:creator>Chu, P.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Kunder, C. A.</dc:creator>
<dc:creator>Petrov, D. A.</dc:creator>
<dc:creator>Winslow, M. M.</dc:creator>
<dc:date>2021-10-21</dc:date>
<dc:identifier>doi:10.1101/2021.10.20.464849</dc:identifier>
<dc:title><![CDATA[Combinatorial tumor suppressor inactivation efficiently initiates lung adenocarcinoma with therapeutic vulnerabilities]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.10.22.465384v1?rss=1">
<title>
<![CDATA[
In-vitro cellular reprogramming to model gonad development and its disorders 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.10.22.465384v1?rss=1"
</link>
<description><![CDATA[
During embryonic development, mutually antagonistic signaling cascades determine the fate of the bipotential gonad towards a testicular or ovarian identity. Errors in this process result in human Disorders of Sex Development (DSDs), where there is discordance between chromosomal, gonadal, and anatomical sex. The absence of an appropriate, accessible in-vitro system is a major obstacle in understanding mechanisms of sex-determination/DSDs. Here, we describe protocols for differentiation of mouse and human pluripotent cells towards gonadal progenitors. Transcriptomic analysis reveals that the in-vitro-derived murine gonadal cells are equivalent to E11.5 in-vivo progenitors. Using similar conditions, Sertoli-like cells derived from 46,XY human induced pluripotent stem cells (hiPSCs) exhibit sustained expression of testis-specific genes, secrete AMH, migrate and form tubular structures. The cells derived from a 46,XY DSD female hiPSCs, carrying a NR5A1 variant, show aberrant gene expression and absence of tubule formation. CRISPR/Cas9-mediated correction of the variant rescued the phenotype. This is a robust tool to understand mechanisms of sex-determination and model DSDs.
]]></description>
<dc:creator>Gonen, N.</dc:creator>
<dc:creator>Eozenou, C.</dc:creator>
<dc:creator>Mitter, R.</dc:creator>
<dc:creator>Bernardo, A.</dc:creator>
<dc:creator>Chervova, A.</dc:creator>
<dc:creator>Frachon, E.</dc:creator>
<dc:creator>Commere, P.-H.</dc:creator>
<dc:creator>Mazen, I.</dc:creator>
<dc:creator>Gobaa, S.</dc:creator>
<dc:creator>McElreavey, K.</dc:creator>
<dc:creator>Lovell-Badge, R.</dc:creator>
<dc:creator>Bashamboo, A.</dc:creator>
<dc:date>2021-10-24</dc:date>
<dc:identifier>doi:10.1101/2021.10.22.465384</dc:identifier>
<dc:title><![CDATA[In-vitro cellular reprogramming to model gonad development and its disorders]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.10.24.465596v1?rss=1">
<title>
<![CDATA[
ZELDA: a 3D Image Segmentation and Parent-Child relation plugin for microscopy image analysis in napari 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.10.24.465596v1?rss=1"
</link>
<description><![CDATA[
Bioimage analysis workflows allow the measurement of sample properties such as fluorescence intensity and polarization, cell number, and vesicles distribution, but often require the integration of multiple software tools. Furthermore, it is increasingly appreciated that to overcome the limitations of the 2D-view-based image analysis approaches and to correctly understand and interpret biological processes, a 3D segmentation of microscopy data sets becomes imperative.

Despite the availability of numerous algorithms for the 2D and 3D segmentation, the latter still offers some challenges for the end-users, who often do not have either an extensive knowledge of the existing software or coding skills to link the output of multiple tools. While several commercial packages are available on the market, fewer are the open-source solutions able to execute a complete 3D analysis workflow.

Here we present ZELDA, a new napari plugin that easily integrates the cutting-edge solutions offered by python ecosystem, such as scikit-image for image segmentation, matplotlib for data visualization, and napari multi-dimensional image viewer for 3D rendering. This plugin aims to provide interactive and zero-scripting customizable workflows for cell segmentation, vesicles counting, parent-child relation between objects, signal quantification, and results presentation; all included in the same open-source napari viewer, and "few clicks away".
]]></description>
<dc:creator>D'Antuono, R.</dc:creator>
<dc:creator>Pisignano, G.</dc:creator>
<dc:date>2021-10-24</dc:date>
<dc:identifier>doi:10.1101/2021.10.24.465596</dc:identifier>
<dc:title><![CDATA[ZELDA: a 3D Image Segmentation and Parent-Child relation plugin for microscopy image analysis in napari]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.10.24.464690v1?rss=1">
<title>
<![CDATA[
The proteasome regulator PSME4 drives immune evasion and abrogates anti-tumor immunity in NSCLC 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.10.24.464690v1?rss=1"
</link>
<description><![CDATA[
Protein degradation by proteasomes is important for the immune response against tumors. Antigens generated by the proteasome promote immune cell infiltration into tumors and improve tumors responses to immunotherapy. For example, immunoproteasomes - a subset of proteasomes induced by inflammatory signals - may improve the response of melanomas to immune checkpoint inhibitors (ICI) by eliciting tumor inflammation. Yet, it is unclear whether and how protein degradation by proteasomes impacts cancer progression and contributes to immune evasion and resistance. Here, we profile the proteasome-cleaved peptides in lung cancers and find that PSME4 serves as a novel inhibitory regulator of the immunoproteasome, playing an anti-inflammatory role in cancer. Biochemical assays combined with scRNA-seq, immunopeptidomics and in vivo analyses demonstrate that PSME4 promotes an immunosuppressive environment around the tumor and abrogates anti-tumor immunity by inhibiting antigen presentation and attenuating tumor inflammation. Furthermore, we find that PSME4 expression is correlated with responsiveness to ICI across several cancer types. Our findings suggest that PSME4-mediated regulation of proteasome activity is a novel mechanism of immune evasion in non-small-cell lung carcinoma and may be targeted therapeutically for restoring anti-tumor immunity.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=139 SRC="FIGDIR/small/464690v1_ufig1.gif" ALT="Figure 1">
View larger version (39K):
org.highwire.dtl.DTLVardef@e87910org.highwire.dtl.DTLVardef@641843org.highwire.dtl.DTLVardef@175092corg.highwire.dtl.DTLVardef@ad34c2_HPS_FORMAT_FIGEXP  M_FIG C_FIG HighlightsO_LIMapping the degradation landscape in Non-Small Cell Lung Cancer (NSCLC) uncovers altered proteasome activity and composition
C_LIO_LIProteasome regulator PSME4 plays an anti-inflammatory role in NSCLC by attenuating immunoproteasome activity
C_LIO_LIPSME4 restricts tumor antigen presentation and cytokine secretion, defining a  cold tumor environment
C_LIO_LIPSME4 drives tumor immune evasion and is associated with resistance to immunotherapy
C_LI
]]></description>
<dc:creator>Javitt, A.</dc:creator>
<dc:creator>Shmueli, M. D.</dc:creator>
<dc:creator>Kramer, M. P.</dc:creator>
<dc:creator>Kolodziejczyk, A. A.</dc:creator>
<dc:creator>Cohen, I. J.</dc:creator>
<dc:creator>Kamer, I.</dc:creator>
<dc:creator>Litchfield, K.</dc:creator>
<dc:creator>Bab-Dinitz, E.</dc:creator>
<dc:creator>Zadok, O.</dc:creator>
<dc:creator>Neiens, V.</dc:creator>
<dc:creator>Ulman, A.</dc:creator>
<dc:creator>Radomir, L.</dc:creator>
<dc:creator>Wolf-Levy, H.</dc:creator>
<dc:creator>Eisenberg-Lerner, A.</dc:creator>
<dc:creator>Kacen, A.</dc:creator>
<dc:creator>Alon, M.</dc:creator>
<dc:creator>Toste Rego, A.</dc:creator>
<dc:creator>Stacher-Priehse, E.</dc:creator>
<dc:creator>Linder, M.</dc:creator>
<dc:creator>Koch, I.</dc:creator>
<dc:creator>Bar, J.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Samuels, Y.</dc:creator>
<dc:creator>Levin, Y.</dc:creator>
<dc:creator>C. A. da Fonseca, P.</dc:creator>
<dc:creator>Elinav, E.</dc:creator>
<dc:creator>Friedman, N.</dc:creator>
<dc:creator>Meiners, S.</dc:creator>
<dc:creator>Merbl, Y.</dc:creator>
<dc:date>2021-10-24</dc:date>
<dc:identifier>doi:10.1101/2021.10.24.464690</dc:identifier>
<dc:title><![CDATA[The proteasome regulator PSME4 drives immune evasion and abrogates anti-tumor immunity in NSCLC]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.10.27.465188v1?rss=1">
<title>
<![CDATA[
RNA sensing via LGP2 is essential for the induction of a type I IFN response in ADAR1 deficiency 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.10.27.465188v1?rss=1"
</link>
<description><![CDATA[
RNA editing by the enzyme Adenosine Deaminase Acting on RNA 1 (ADAR1) is an important mechanism by which cells avoid innate immune responses to some endogenous RNAs. In ADAR1-deficient cells, unedited self RNAs can form base-paired structures that resemble viral RNAs and inadvertently activate antiviral innate immune pathways that lead to the induction of type I interferon (IFN). Rare mutations in ADAR1 cause Aicardi-Goutieres Syndrome (AGS), a severe childhood autoinflammatory syndrome that is characterized by chronic and excessive type I IFN production and developmental delay. Conversely, ADAR1 dysfunction and consequent type I IFN production helps restrict tumor growth and potentiates the activity of some chemotherapy drugs. Induction of type I IFN in ADAR1-deficient cells is thought to be due to triggering of the cytosolic RIG-I-like receptor (RLR), MDA5, by unedited self RNAs. Here, we show that another RLR, LGP2, also has an essential role. We demonstrate that ADAR1-deficient human cells fail to mount a type I IFN response in the absence of LGP2 and this involves the canonical function of LGP2 as an RNA sensor and facilitator of MDA5-dependent signaling. Further, we show that the sensitivity of tumor cells to ADAR1 loss requires the presence of LGP2. Finally, we find that type I IFN induction in tumor cells depleted of ADAR1 and treated with some chemotherapeutics is fully dependent on the expression of LGP2. These findings highlight a central role for LGP2 in self RNA sensing with important clinical implications for the treatment of AGS as well as for the potential application of ADAR1-directed anti-tumor therapy.
]]></description>
<dc:creator>Stok, J. E.</dc:creator>
<dc:creator>Oosenbrug, T.</dc:creator>
<dc:creator>ter Haar, L. R.</dc:creator>
<dc:creator>Gravekamp, D.</dc:creator>
<dc:creator>Bromley, C. P.</dc:creator>
<dc:creator>Zelenay, S.</dc:creator>
<dc:creator>Reis e Sousa, C.</dc:creator>
<dc:creator>van der Veen, A. G.</dc:creator>
<dc:date>2021-10-28</dc:date>
<dc:identifier>doi:10.1101/2021.10.27.465188</dc:identifier>
<dc:title><![CDATA[RNA sensing via LGP2 is essential for the induction of a type I IFN response in ADAR1 deficiency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.10.27.465994v1?rss=1">
<title>
<![CDATA[
Chronumental: time tree estimation from very large phylogenies 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.10.27.465994v1?rss=1"
</link>
<description><![CDATA[
Phylogenetic trees are an important tool for interpreting sequenced genomes, and their interrelationships. Estimating the date associated with each node of such a phylogeny creates a "time tree", which can be especially useful for visualising and analysing evolution of organisms such as viruses. Several tools have been developed for time-tree estimation, but the sequencing explosion in response to the SARS-CoV-2 pandemic has created phylogenies so large as to prevent the application of these previous approaches to full datasets. Here we introduce Chronumental, a tool that can rapidly infer time trees from phylogenies featuring large numbers of nodes. Chronumental uses stochastic gradient descent to identify lengths of time for tree branches which maximise the evidence lower bound under a probabilistic model, implemented in a framework which can be compiled into XLA for rapid computation. We show that Chronumental scales to phylogenies featuring millions of nodes, with chronological predictions made in minutes, and is able to accurately predict the dates of nodes for which it is not provided with metadata.
]]></description>
<dc:creator>Sanderson, T.</dc:creator>
<dc:date>2021-10-28</dc:date>
<dc:identifier>doi:10.1101/2021.10.27.465994</dc:identifier>
<dc:title><![CDATA[Chronumental: time tree estimation from very large phylogenies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.11.02.466974v1?rss=1">
<title>
<![CDATA[
N4BP1 is dimerization-dependent linear ubiquitin reader regulating TNFR1 signalling through linear ubiquitin binding and Caspase-8-mediated processing 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.11.02.466974v1?rss=1"
</link>
<description><![CDATA[
Signalling through TNFR1 modulates proinflammatory gene transcription and programmed cell death, and its impairment causes autoimmune diseases and cancer. NEDD4-binding protein 1 (N4BP1) was recently identified as a critical suppressor of proinflammatory cytokine production1, whose mode of action remained unknown. Here, we show that N4BP1 is a novel linear ubiquitin reader that negatively regulates NF{kappa}B signalling by its unique dimerizationdependent ubiquitin-binding module that we named LUBIN. Dimeric N4BP1 strategically positions two non-selective ubiquitin-binding domains to ensure exclusive recognition of linear ubiquitin. Under proinflammatory conditions, N4BP1 is recruited to the nascent TNFR1 signalling complex, where it regulates duration of proinflammatory signalling in LUBIN-dependent manner. N4BP1 deficiency accelerates TNF-induced cell death by increasing complex II assembly. Under proapoptotic conditions, Caspase-8 mediates proteolytic processing of N4BP1 and the resulting cleavage fragment of N4BP1, which retains the ability to bind linear ubiquitin, is rapidly degraded by the 26S proteasome, accelerating apoptosis. In summary, our findings demonstrate that N4BP1 dimerization creates a unique linear ubiquitin reader that ensures timely and coordinated regulation of TNFR1-mediated inflammation and cell death.
]]></description>
<dc:creator>Kliza, K. W.</dc:creator>
<dc:creator>Song, W.</dc:creator>
<dc:creator>Pinzuti, I.</dc:creator>
<dc:creator>Schaubeck, S.</dc:creator>
<dc:creator>Kunzelmann, S.</dc:creator>
<dc:creator>Kuntin, D.</dc:creator>
<dc:creator>Fornili, A.</dc:creator>
<dc:creator>Pandini, A.</dc:creator>
<dc:creator>Hofmann, K.</dc:creator>
<dc:creator>Garnett, J.</dc:creator>
<dc:creator>Stieglitz, B.</dc:creator>
<dc:creator>Husnjak, K.</dc:creator>
<dc:date>2021-11-02</dc:date>
<dc:identifier>doi:10.1101/2021.11.02.466974</dc:identifier>
<dc:title><![CDATA[N4BP1 is dimerization-dependent linear ubiquitin reader regulating TNFR1 signalling through linear ubiquitin binding and Caspase-8-mediated processing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.11.03.467152v1?rss=1">
<title>
<![CDATA[
DeepBacs: Bacterial image analysis using open-source deep learning approaches 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.11.03.467152v1?rss=1"
</link>
<description><![CDATA[
Deep Learning (DL) is rapidly changing the field of microscopy, allowing for efficient analysis of complex data while often out-performing classical algorithms. This revolution has led to a significant effort to create user-friendly tools allowing biomedical researchers with little background in computer sciences to use this technology effectively. Thus far, these approaches have mainly focused on analysing microscopy images from eukaryotic samples and are still underused in microbiology. In this work, we demonstrate how to use a range of state-of-the-art artificial neural-networks particularly suited for the analysis of bacterial microscopy images, using our recently developed ZeroCostDL4Mic platform. We showcase different DL approaches for segmenting bright field and fluorescence images of different bacterial species, use object detection to classify different growth stages in time-lapse imaging data, and carry out DL-assisted phenotypic profiling of antibiotic-treated cells. To also demonstrate the DL capacity to enhance low-phototoxicity live-cell microscopy, we showcase how image denoising can allow researchers to attain high-fidelity data in faster and longer imaging. Finally, artificial labelling of cell membranes and predictions of super-resolution images allow for accurate mapping of cell shape and intracellular targets. To aid in the training of novice users, we provide a purposefully-built database of training and testing data, enabling bacteriologists to quickly explore how to analyse their data through DL. We hope this lays a fertile ground for the efficient application of DL in microbiology and fosters the creation of novel tools for bacterial cell biology and antibiotic research.
]]></description>
<dc:creator>Spahn, C.</dc:creator>
<dc:creator>Laine, R. F.</dc:creator>
<dc:creator>Pereira, P. M.</dc:creator>
<dc:creator>Gomez-de-Mariscal, E.</dc:creator>
<dc:creator>von Chamier, L.</dc:creator>
<dc:creator>Conduit, M.</dc:creator>
<dc:creator>Pinho, M. G.</dc:creator>
<dc:creator>Holden, S.</dc:creator>
<dc:creator>Jacquemet, G.</dc:creator>
<dc:creator>Heilemann, M.</dc:creator>
<dc:creator>Henriques, R.</dc:creator>
<dc:date>2021-11-03</dc:date>
<dc:identifier>doi:10.1101/2021.11.03.467152</dc:identifier>
<dc:title><![CDATA[DeepBacs: Bacterial image analysis using open-source deep learning approaches]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.11.03.467007v1?rss=1">
<title>
<![CDATA[
Increasing the throughput of sensitive proteomics by plexDIA 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.11.03.467007v1?rss=1"
</link>
<description><![CDATA[
Current mass-spectrometry methods enable high-throughput proteomics of large sample amounts, but proteomics of low sample amounts remains limited in depth and throughput. To increase the throughput of sensitive proteomics, we developed an experimental and computational framework, plexDIA, for simultaneously multiplexing the analysis of both peptides and samples. Multiplexed analysis with plexDIA increases throughput multiplicatively with the number of labels without reducing proteome coverage or quantitative accuracy. By using 3-plex nonisobaric mass tags, plexDIA enables quantifying 3-fold more protein ratios among nanogram-level samples. Using 1 hour active gradients and first-generation Q Exactive, plexDIA quantified about 8,000 proteins in each sample of labeled 3-plex sets. plexDIA also increases data completeness, reducing missing data over 2-fold across samples. We applied plexDIA to quantify proteome dynamics during the cell division cycle in cells isolated based on their DNA content; plexDIA detected many classical cell cycle proteins and discovered new ones. When applied to single human cells, plexDIA quantified about 1,000 proteins per cell and achieved 98 % data completeness within a plexDIA set while using about 5 min of active chromatography per cell. These results establish a general framework for increasing the throughput of sensitive and quantitative protein analysis.



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=81 SRC="FIGDIR/small/467007v2_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Derks, J.</dc:creator>
<dc:creator>Leduc, A.</dc:creator>
<dc:creator>Huffman, R. G.</dc:creator>
<dc:creator>Specht, H.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:creator>Demichev, V.</dc:creator>
<dc:creator>Slavov, N.</dc:creator>
<dc:date>2021-11-04</dc:date>
<dc:identifier>doi:10.1101/2021.11.03.467007</dc:identifier>
<dc:title><![CDATA[Increasing the throughput of sensitive proteomics by plexDIA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.11.08.467712v1?rss=1">
<title>
<![CDATA[
The molecular evolution of spermatogenesis across mammals 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.11.08.467712v1?rss=1"
</link>
<description><![CDATA[
The testis is a key male reproductive organ that produces gametes through the process of spermatogenesis. Testis morphologies and spermatogenesis evolve rapidly in mammals, presumably due to the evolutionary pressure on males to be reproductively successful1,2. The rapid evolution of the testis was shown to be reflected at the molecular level based on bulk-tissue work3-8, but the molecular evolution of individual spermatogenic cell types across mammalian lineages remains largely uncharacterized. Here we report evolutionary analyses of single-nucleus transcriptome data for testes from eleven species that cover the three major mammalian lineages (eutherians, marsupials, egg-laying monotremes) and birds (the evolutionary outgroup), and include seven key primates. Our analyses reveal that the rapid evolution of the testis is driven by accelerated fixation rates of gene expression changes, amino acid altering substitutions, and newly emerged genes in late spermatogenic stages - likely facilitated by reduced pleiotropic constraints, haploid selection, and a transcriptionally permissive chromatin environment. We identify temporal expression changes of individual genes across species, which may have contributed to the emergence of species-specific phenotypes, but also conserved expression programs underlying ancestral spermatogenic processes. Sex chromosome analyses show that genes predominantly expressed in spermatogonia (i.e., germ cells fueling spermatogenesis) and Sertoli cells (i.e., somatic supporting cells) independently accumulated on X chromosomes across mammals during evolution, presumably due to male-beneficial selective forces. Further work uncovered that the process of meiotic sex chromosome inactivation (MSCI) also occurs in monotremes and hence is common to the different mammalian sex chromosome systems, contrary to previous inferences9. Thus, the general mechanism of meiotic silencing of unsynapsed chromatin (MSUC), which underlies MSCI, represents an ancestral mammalian feature. Together, our study illuminates the cellular and molecular evolution of mammalian spermatogenesis and associated selective forces, and provides a resource for investigating the biology of the testis across mammals.
]]></description>
<dc:creator>Murat, F.</dc:creator>
<dc:creator>Mbengue, N.</dc:creator>
<dc:creator>Winge, S. B.</dc:creator>
<dc:creator>Trefzer, T.</dc:creator>
<dc:creator>Leushkin, E.</dc:creator>
<dc:creator>Sepp, M.</dc:creator>
<dc:creator>Cardoso-Moreira, M.</dc:creator>
<dc:creator>Schmidt, J.</dc:creator>
<dc:creator>Schneider, C.</dc:creator>
<dc:creator>Mössinger, K.</dc:creator>
<dc:creator>Brüning, T.</dc:creator>
<dc:creator>Lamanna, F.</dc:creator>
<dc:creator>Belles, M. R.</dc:creator>
<dc:creator>Conrad, C.</dc:creator>
<dc:creator>Kondova, I.</dc:creator>
<dc:creator>Bontrop, R.</dc:creator>
<dc:creator>Behr, R.</dc:creator>
<dc:creator>Khaitovich, P.</dc:creator>
<dc:creator>Pääbo, S.</dc:creator>
<dc:creator>Marques-Bonet, T.</dc:creator>
<dc:creator>Grützner, F.</dc:creator>
<dc:creator>Almstrup, K.</dc:creator>
<dc:creator>Schierup, M. H.</dc:creator>
<dc:creator>Kaessmann, H.</dc:creator>
<dc:date>2021-11-08</dc:date>
<dc:identifier>doi:10.1101/2021.11.08.467712</dc:identifier>
<dc:title><![CDATA[The molecular evolution of spermatogenesis across mammals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.11.11.468272v1?rss=1">
<title>
<![CDATA[
PHGDH is required for germinal center formation and is a therapeutic target in MYC-driven lymphoma 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.11.11.468272v1?rss=1"
</link>
<description><![CDATA[
The synthesis of serine from glucose is a key metabolic pathway supporting cellular proliferation in healthy and malignant cells. Despite this, the role that this aspect of metabolism plays in germinal center biology and pathology is not known. Here, we performed a comprehensive characterization of the role of the serine synthesis pathway in germinal center B cells and lymphomas derived from these cells. We demonstrate that upregulation of a functional serine synthesis pathway is a metabolic hallmark of B-cell activation and the germinal center reaction. Inhibition of phosphoglycerate dehydrogenase (PHGDH), the first and rate limiting enzyme in this pathway, leads to defective germinal formation and impaired high-affinity antibody production. In addition, overexpression of enzymes involved in serine synthesis is a characteristic of germinal center B-cell derived lymphomas, with high levels of expression being predictive of reduced overall survival in diffuse large B cell lymphoma. Inhibition of PHGDH induces apoptosis in lymphoma cells reducing disease progression. These findings establish PHGDH as a critical player in humoral immunity and a clinically relevant target in lymphoma.
]]></description>
<dc:creator>D'Avola, A.</dc:creator>
<dc:creator>Legrave, N.</dc:creator>
<dc:creator>Tajan, M.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Shearer, R.</dc:creator>
<dc:creator>King, H.</dc:creator>
<dc:creator>Cheung, E.</dc:creator>
<dc:creator>Clear, A.</dc:creator>
<dc:creator>Gunawan, A.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>James, L.</dc:creator>
<dc:creator>MacRae, J.</dc:creator>
<dc:creator>Gribben, J.</dc:creator>
<dc:creator>Calado, D.</dc:creator>
<dc:creator>Vousden, K.</dc:creator>
<dc:creator>Riches, J.</dc:creator>
<dc:date>2021-11-12</dc:date>
<dc:identifier>doi:10.1101/2021.11.11.468272</dc:identifier>
<dc:title><![CDATA[PHGDH is required for germinal center formation and is a therapeutic target in MYC-driven lymphoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.11.15.468579v1?rss=1">
<title>
<![CDATA[
PPAR-delta acts as a metabolic master checkpoint for metastasis in pancreatic cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.11.15.468579v1?rss=1"
</link>
<description><![CDATA[
SO_SCPLOWUMMARYC_SCPLOWIn pancreatic cancer, emerging evidence suggests that PPAR-{delta} overexpression is associated with tumor progression and metastasis, but a mechanistic link is still missing. Here we now show that PPAR-{delta} acts as the integrating upstream regulator for the metabolic rewiring, which is preceding the subsequent initiation of an invasive/metastatic program. Specifically, paracrine and metabolic cues regularly found in the metastasis-promoting tumor stroma consistently enhance, via induction of PPAR-{delta} activity, the glycolytic capacity and reserve of pancreatic cancer cells, respectively, accompanied by decreased mitochondrial oxygen consumption. Consequently, genetic or pharmacological inhibition of PPAR-{delta} results in reduced invasiveness and metastasis. Mechanistically, PPAR-{delta} acts by shifting the MYC/PGC1A balance towards MYC, enhancing metabolic plasticity. Targeting MYC similarly prevents the metabolic switch and subsequent initiation of invasiveness. Therefore, our data demonstrate that PPAR-{delta} is a key initiator for the metabolic reprogramming in pancreatic cancer, thereby acting as a checkpoint for the phenotypic change towards invasiveness. These findings provide compelling evidence for a novel treatment strategy to combat pancreatic cancer progression.
]]></description>
<dc:creator>Parejo-Alonso, B.</dc:creator>
<dc:creator>Barneda, D.</dc:creator>
<dc:creator>Trabulo, S. M.</dc:creator>
<dc:creator>Courtois, S.</dc:creator>
<dc:creator>Compte-Sancerni, S.</dc:creator>
<dc:creator>Ruiz-Canas, L.</dc:creator>
<dc:creator>Zheng, Q.</dc:creator>
<dc:creator>Tang, J.</dc:creator>
<dc:creator>Chen, M.</dc:creator>
<dc:creator>Guo, Z.</dc:creator>
<dc:creator>Schmitz, U.</dc:creator>
<dc:creator>Irun, P.</dc:creator>
<dc:creator>Penin-Peyta, L.</dc:creator>
<dc:creator>Crusz, S. M.</dc:creator>
<dc:creator>Cano-Galiano, A.</dc:creator>
<dc:creator>Lopez-Escalona, S.</dc:creator>
<dc:creator>Jagust, P.</dc:creator>
<dc:creator>Espiau-Romera, P.</dc:creator>
<dc:creator>Yuneva, M.</dc:creator>
<dc:creator>Lin, M.-L.</dc:creator>
<dc:creator>Lanas, A.</dc:creator>
<dc:creator>Sainz, B.</dc:creator>
<dc:creator>Heeschen, C.</dc:creator>
<dc:creator>Sancho, P.</dc:creator>
<dc:date>2021-11-16</dc:date>
<dc:identifier>doi:10.1101/2021.11.15.468579</dc:identifier>
<dc:title><![CDATA[PPAR-delta acts as a metabolic master checkpoint for metastasis in pancreatic cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.11.24.469674v1?rss=1">
<title>
<![CDATA[
Interplay of adherens junctions and matrix proteolysis determines the invasive pattern and growth of squamous cell carcinoma 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.11.24.469674v1?rss=1"
</link>
<description><![CDATA[
Cancers, such as squamous cell carcinoma, frequently invade as multicellular units. However, these invading units can be organized in a variety of ways, ranging from thin discontinuous strands to thick  pushing collectives. Here we employ an integrated experimental and computational approach to identify the factors that determine the mode of collective cancer cell invasion. We find that matrix proteolysis is linked to the formation of wide strands, but has little effect on the maximum extent of invasion. Cell-cell junctions also favour wide strands, but our analysis also reveals a requirement for cell-cell junctions for efficient invasion in response to uniform directional cues. Unexpectedly, the ability to generate wide invasive strands is coupled to the ability to grow effectively when surrounded by ECM in 3D assays. Combinatorial perturbation of both matrix proteolysis and cell-cell adhesion demonstrates that the most aggressive cancer behaviour, both in terms of invasion and growth, is achieved at high levels of cell-cell adhesion and high levels of proteolysis. Contrary to expectation, cells with canonical mesenchymal traits - no cell-cell junctions and high proteolysis - exhibit reduced growth and lymph node metastasis. Thus, we conclude that the ability of squamous cell carcinoma cells to invade effectively is also linked to their ability to generate space for proliferation in confined contexts. These data provide an explanation for the apparent advantage of retaining cell-cell junctions in SCC.
]]></description>
<dc:creator>Kato, T.</dc:creator>
<dc:creator>Jenkins, R. P.</dc:creator>
<dc:creator>Derzsi, S.</dc:creator>
<dc:creator>Tozluoglu, M.</dc:creator>
<dc:creator>Rullan, A.</dc:creator>
<dc:creator>Hooper, S.</dc:creator>
<dc:creator>Chaleil, R. A. G.</dc:creator>
<dc:creator>Fu, X.</dc:creator>
<dc:creator>Joyce, H.</dc:creator>
<dc:creator>Thavaraj, S.</dc:creator>
<dc:creator>Bates, P. A.</dc:creator>
<dc:creator>Sahai, E.</dc:creator>
<dc:date>2021-11-24</dc:date>
<dc:identifier>doi:10.1101/2021.11.24.469674</dc:identifier>
<dc:title><![CDATA[Interplay of adherens junctions and matrix proteolysis determines the invasive pattern and growth of squamous cell carcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.11.29.470317v1?rss=1">
<title>
<![CDATA[
A positive feedback loop mediates crosstalk between calcium, cyclic nucleotide and lipid signalling in Toxoplasma gondii 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.11.29.470317v1?rss=1"
</link>
<description><![CDATA[
Fundamental processes of obligate intracellular parasites, such as Toxoplasma gondii and Plasmodium falciparum, are controlled by a set of plant-like calcium dependent kinases (CDPKs), the conserved cAMP- and cGMP-dependent protein kinases (PKA and PKG), secondary messengers and lipid signalling. While some major components of the signalling networks have been identified, how these are connected remains largely unknown. Here, we compare the phospho-signalling networks during Toxoplasma egress from its host cell by artificially raising cGMP or calcium levels to activate PKG or CDPKs, respectively. We show that both these inducers trigger near identical signalling pathways and provide evidence for a positive feedback loop involving CDPK3. We measure phospho- and lipid signalling in parasites treated with the Ca2+ ionophore A23187 in a sub-minute timecourse and show CDPK3-dependent regulation of diacylglycerol levels and increased phosphorylation of four phosphodiesterases (PDEs), suggesting their function in the feedback loop. Disruption of CDPK3 leads to elevated cAMP levels and inhibition of PKA signalling rescues the egress defect of {Delta}CDPK3 parasites treated with A23187. Biochemical analysis of the four PDEs identifies PDE2 as the only cAMP-specific PDE among these candidates, while the other PDEs are cGMP specific, two of which are inhibited by the predicted PDE inhibitor BIPPO. Conditional deletion of the four PDEs supports an important, but non-essential role for PDE1 and PDE2 in growth, with PDE2 controlling A23187-mediated egress. In summary we uncover a positive feedback loop that enhances signalling during egress and links several signalling pathways together.
]]></description>
<dc:creator>Dominicus, C.</dc:creator>
<dc:creator>Nofal, S. D.</dc:creator>
<dc:creator>Broncel, M.</dc:creator>
<dc:creator>Kastris, N. J.</dc:creator>
<dc:creator>Flynn, H.</dc:creator>
<dc:creator>Arrizabalaga, G.</dc:creator>
<dc:creator>Botte, C. Y.</dc:creator>
<dc:creator>Invergo, B. M.</dc:creator>
<dc:creator>Treeck, M.</dc:creator>
<dc:date>2021-11-30</dc:date>
<dc:identifier>doi:10.1101/2021.11.29.470317</dc:identifier>
<dc:title><![CDATA[A positive feedback loop mediates crosstalk between calcium, cyclic nucleotide and lipid signalling in Toxoplasma gondii]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.11.30.470568v1?rss=1">
<title>
<![CDATA[
Broad human and animal coronavirus neutralisation by SARS-CoV-2 S2-targeted vaccination 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.11.30.470568v1?rss=1"
</link>
<description><![CDATA[
Several common-cold coronaviruses (HCoVs) are endemic in humans and several variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have emerged during the current Coronavirus disease 2019 (COVID-19) pandemic. Whilst antibody cross-reactivity with the Spike glycoproteins (S) of diverse coronaviruses has been documented, it remains unclear whether such antibody responses, typically targeting the conserved S2 subunit, contribute to or mediate protection, when induced naturally or through vaccination. Using a mouse model, we show that prior HCoV-OC43 S immunity primes neutralising antibody responses to otherwise subimmunogenic SARS-CoV-2 S exposure and promotes S2-targeting antibody responses. Moreover, mouse vaccination with SARS-CoV-2 S2 elicits antibodies that neutralise diverse animal and human alphacoronaviruses and betacoronaviruses in vitro, and protects against SARS-CoV-2 challenge in vivo. Lastly, in mice with a history of SARS-CoV-2 Wuhan-based S vaccination, further S2 vaccination induces stronger and broader neutralising antibody response than booster Wuhan S vaccination, suggesting it may prevent repertoire focusing caused by repeated homologous vaccination. The data presented here establish the protective value of an S2-targeting vaccine and support the notion that S2 vaccination may better prepare the immune system to respond to the changing nature of the S1 subunit in SARS-CoV-2 variants of concern (VOCs), as well as to unpredictable, yet inevitable future coronavirus zoonoses.
]]></description>
<dc:creator>Ng, K.</dc:creator>
<dc:creator>Faulkner, N.</dc:creator>
<dc:creator>Finsterbusch, K.</dc:creator>
<dc:creator>Wu, M.</dc:creator>
<dc:creator>Harvey, R.</dc:creator>
<dc:creator>Hussain, S.</dc:creator>
<dc:creator>Greco, M.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Kjaer, S.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Beale, R.</dc:creator>
<dc:creator>Gamblin, S.</dc:creator>
<dc:creator>Cherepanov, P.</dc:creator>
<dc:creator>Mccauley, J.</dc:creator>
<dc:creator>Daniels, R.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>Arase, H.</dc:creator>
<dc:creator>Wack, A.</dc:creator>
<dc:creator>Bauer, D.</dc:creator>
<dc:creator>Kassiotis, G.</dc:creator>
<dc:date>2021-12-01</dc:date>
<dc:identifier>doi:10.1101/2021.11.30.470568</dc:identifier>
<dc:title><![CDATA[Broad human and animal coronavirus neutralisation by SARS-CoV-2 S2-targeted vaccination]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.12.03.471138v1?rss=1">
<title>
<![CDATA[
Plasmodium SAS4/CPAP is a flagellum basal body component during male gametogenesis but is not essential for parasite transmission 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.12.03.471138v1?rss=1"
</link>
<description><![CDATA[
The centriole/basal body (CBB) is an evolutionarily conserved organelle acting as a microtubule organising centre (MTOC) to nucleate cilia, flagella and the centrosome. SAS4/CPAP is a conserved component associated with BB biogenesis in many model flagellated cells. Plasmodium, a divergent unicellular eukaryote and causative agent of malaria, displays an atypical closed mitosis with an MTOC, reminiscent of the acentriolar MTOC, embedded in the nuclear membrane at most proliferative stages. Mitosis during male gamete formation is accompanied by flagellum formation: within 15 minutes, genome replication (from 1N to 8N) and three successive rounds of mitosis without nuclear division occur, with coordinated axoneme biogenesis in the cytoplasm resulting in eight flagellated gametes. There are two MTOCs in male gametocytes. An acentriolar MTOC located with the nuclear envelope and a centriolar MTOC (basal body) located within the cytoplasm that are required for flagellum assembly. To study the location and function of SAS4 during this rapid process, we examined the spatial profile of SAS4 in real time by live cell imaging and its function by gene deletion. We show its absence during asexual proliferation but its presence and coordinated association and assembly of SAS4 with another basal body component, kinesin8B, which is involved in axoneme biogenesis. In contrast its separation from the nuclear kinetochore marker NDC80 suggests that SAS4 is part of the basal body and outer centriolar MTOC residing in the cytoplasm. However, deletion of the SAS4 gene produced no phenotype, indicating that it is not essential for male gamete formation or parasite transmission through the mosquito.
]]></description>
<dc:creator>Zeeshan, M.</dc:creator>
<dc:creator>Brady, D.</dc:creator>
<dc:creator>Markus, R.</dc:creator>
<dc:creator>Vaughan, S.</dc:creator>
<dc:creator>Ferguson, D.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:creator>Tewari, R.</dc:creator>
<dc:date>2021-12-03</dc:date>
<dc:identifier>doi:10.1101/2021.12.03.471138</dc:identifier>
<dc:title><![CDATA[Plasmodium SAS4/CPAP is a flagellum basal body component during male gametogenesis but is not essential for parasite transmission]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.12.07.471544v1?rss=1">
<title>
<![CDATA[
Positional motif analysis reveals the extent of specificity of protein-RNA interactions observed by CLIP 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.12.07.471544v1?rss=1"
</link>
<description><![CDATA[
BackgroundCrosslinking and immunoprecipitation (CLIP) is a method used to identify in vivo RNA- protein binding sites on a transcriptome-wide scale. With the increasing amounts of available data for RNA-binding proteins (RBPs), it is important to understand to what degree the enriched motifs specify the RNA binding profiles of RBPs in cells.

ResultsWe develop positionally-enriched k-mer analysis (PEKA), a computational tool for efficient analysis of enriched motifs from individual CLIP datasets, which minimises the impact of technical and regional genomic biases by internal data normalisation. We cross-validate PEKA with mCross, and show that background correction by size-matched input doesnt generally improve the specificity of detected motifs. We identify motif classes with common enrichment patterns across eCLIP datasets and across RNA regions, while also observing variations in the specificity and the extent of motif enrichment across eCLIP datasets, between variant CLIP protocols, and between CLIP and in vitro binding data. Thereby we gain insights into the contributions of technical and regional genomic biases to the enriched motifs, and find how motif enrichment features relate to the domain composition and low-complexity regions (LCRs) of the studied proteins.

ConclusionsOur study provides insights into the overall contributions of regional binding preferences, protein domains and LCRs to the specificity of protein-RNA interactions, and shows the value of cross-motif and cross-RBP comparison for data interpretation. Our results are presented for exploratory analysis via an online platform in an RBP-centric and motif-centric manner (https://imaps.goodwright.com/apps/peka/). PEKA is available from https://github.com/ulelab/peka.
]]></description>
<dc:creator>Kuret, K.</dc:creator>
<dc:creator>Amalietti, A. G.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:date>2021-12-07</dc:date>
<dc:identifier>doi:10.1101/2021.12.07.471544</dc:identifier>
<dc:title><![CDATA[Positional motif analysis reveals the extent of specificity of protein-RNA interactions observed by CLIP]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.12.09.471958v1?rss=1">
<title>
<![CDATA[
Fission of megamitochondria into multiple smaller well-defined mitochondria in the ageing zebrafish retina 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.12.09.471958v1?rss=1"
</link>
<description><![CDATA[
Mitochondria are essential adenosine triphosphate (ATP)-generating cellular organelles. In the retina, they are highly numerous in the photoreceptors and retinal pigment epithelium (RPE) due to their high energetic requirements. Fission and fusion of the mitochondria within these cells allow them to adapt to changing demands over the lifespan of the organism. Using transmission electron microscopy, we examined the mitochondrial ultrastructure of zebrafish photoreceptors and RPE from 5 days post fertilisation (dpf) through to late adulthood (3 years). Notably, mitochondria in the youngest animals were large and irregular shaped with a loose cristae architecture, but by 8 dpf they had reduced in size and expanded in number with more defined cristae. When investigating temporal gene expression of several mitochondrial-related markers, they indicated fission as the dominant mechanism contributing to these changes observed over time. This is likely to be due to continued mitochondrial stress resulting from the oxidative environment of the retina and prolonged light exposure. We have characterised retinal mitochondrial ageing in a key vertebrate model organism, that provides a basis for future studies of retinal diseases that are linked to mitochondrial dysfunction.
]]></description>
<dc:creator>Burgoyne, T.</dc:creator>
<dc:creator>Toms, M.</dc:creator>
<dc:creator>Way, C.</dc:creator>
<dc:creator>Tracey-White, D.</dc:creator>
<dc:creator>Futter, C.</dc:creator>
<dc:creator>Moosajee, M.</dc:creator>
<dc:date>2021-12-10</dc:date>
<dc:identifier>doi:10.1101/2021.12.09.471958</dc:identifier>
<dc:title><![CDATA[Fission of megamitochondria into multiple smaller well-defined mitochondria in the ageing zebrafish retina]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.12.20.473466v1?rss=1">
<title>
<![CDATA[
Visualizing pyrazinamide action by live single cell imaging of phagosome acidification and Mycobacterium tuberculosis pH homeostasis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.12.20.473466v1?rss=1"
</link>
<description><![CDATA[
The intracellular population of Mycobacterium tuberculosis (Mtb) is dynamically segregated within multiple subcellular niches with different biochemical and biophysical properties that, upon treatment, may impact antibiotic distribution, accumulation, and efficacy. However, it remains unclear whether fluctuating intracellular microenvironments alter mycobacterial homeostasis and contribute to antibiotic enrichment and efficacy. Here, we describe a dual-imaging approach that allows quantitative monitoring of host subcellular acidification and Mtb intrabacterial pH profiles by live-fluorescence microscopy in a biosafety level 3 laboratory. By combining this live imaging approach with pharmacological and genetic perturbations, we show that Mtb can maintain its intracellular pH independently of the surrounding pH in primary human macrophages. Importantly, we show that unlike bedaquiline (BDQ), isoniazid (INH) or rifampicin (RIF), the front-line drug pyrazinamide (PZA) displays antibacterial efficacy by acting as protonophore which disrupts intrabacterial pH homeostasis in cellulo. By using Mtb mutants with different intra-macrophage localisation, we confirmed that intracellular acidification is a prerequisite for PZA efficacy in cellulo. We anticipate this dual imaging approach will be useful to identify host cellular environments that affect antibiotic efficacy against intracellular pathogens.

HighlightsO_LIMtb maintains its intrabacterial pH inside both acidic and neutral subcellular microenvironments of human macrophages
C_LIO_LIPyrazinamide, but not other frontline antibiotics, acts as a protonophore in cellulo
C_LIO_LIPyrazinamide-mediated intrabacterial pH homeostasis disruption and antibacterial efficacy requires host endolysosomal acidification
C_LIO_LICytosolic localisation mediated by ESX-1 contributes to pyrazinamide antibacterial activity resistance
C_LIO_LIPyrazinamide conversion into pyrazinoic acid by the pyrazinamidase/nicotinamidase PncA is essential for its protonophore activity and efficacy in cellulo
C_LI
]]></description>
<dc:creator>Santucci, P.</dc:creator>
<dc:creator>Aylan, B.</dc:creator>
<dc:creator>Botella, L.</dc:creator>
<dc:creator>Bernard, E. M.</dc:creator>
<dc:creator>Bussi, C.</dc:creator>
<dc:creator>Pellegrino, E.</dc:creator>
<dc:creator>Athanasiadi, N.</dc:creator>
<dc:creator>Gutierrez, M. G.</dc:creator>
<dc:date>2021-12-20</dc:date>
<dc:identifier>doi:10.1101/2021.12.20.473466</dc:identifier>
<dc:title><![CDATA[Visualizing pyrazinamide action by live single cell imaging of phagosome acidification and Mycobacterium tuberculosis pH homeostasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.12.20.473443v1?rss=1">
<title>
<![CDATA[
Cellular development and evolution of the mammalian cerebellum 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.12.20.473443v1?rss=1"
</link>
<description><![CDATA[
The expansion of the neocortex, one of the hallmarks of mammalian evolution1,2, was accompanied by an increase in the number of cerebellar neurons3. However, little is known about the evolution of the cellular programs underlying cerebellum development in mammals. In this study, we generated single-nucleus RNA-sequencing data for [~]400,000 cells to trace the development of the cerebellum from early neurogenesis to adulthood in human, mouse, and the marsupial opossum. Our cross-species analyses revealed that the cellular composition and differentiation dynamics throughout cerebellum development are largely conserved, except for human Purkinje cells. Global transcriptome profiles, conserved cell state markers, and gene expression trajectories across neuronal differentiation show that the cerebellar cell type-defining programs have been overall preserved for at least 160 million years. However, we also discovered differences. We identified 3,586 genes that either gained or lost expression in cerebellar cells in one of the species, and 541 genes that evolved new expression trajectories during neuronal differentiation. The potential functional relevance of these cross-species differences is highlighted by the diverged expression patterns of several human disease-associated genes. Altogether, our study reveals shared and lineage-specific programs governing the cellular development of the mammalian cerebellum, and expands our understanding of the evolution of mammalian organ development.
]]></description>
<dc:creator>Sepp, M.</dc:creator>
<dc:creator>Leiss, K.</dc:creator>
<dc:creator>Sarropoulos, I.</dc:creator>
<dc:creator>Murat, F.</dc:creator>
<dc:creator>Okonechnikov, K.</dc:creator>
<dc:creator>Joshi, P.</dc:creator>
<dc:creator>Leushkin, E.</dc:creator>
<dc:creator>Mbengue, N.</dc:creator>
<dc:creator>Schneider, C.</dc:creator>
<dc:creator>Schmidt, J.</dc:creator>
<dc:creator>Trost, N.</dc:creator>
<dc:creator>Spaenig, L.</dc:creator>
<dc:creator>Giere, P.</dc:creator>
<dc:creator>Khaitovich, P.</dc:creator>
<dc:creator>Lisgo, S.</dc:creator>
<dc:creator>Palkovits, M.</dc:creator>
<dc:creator>Kutscher, L. M.</dc:creator>
<dc:creator>Anders, S.</dc:creator>
<dc:creator>Cardoso-Moreira, M.</dc:creator>
<dc:creator>Pfister, S. M.</dc:creator>
<dc:creator>Kaessmann, H.</dc:creator>
<dc:date>2021-12-21</dc:date>
<dc:identifier>doi:10.1101/2021.12.20.473443</dc:identifier>
<dc:title><![CDATA[Cellular development and evolution of the mammalian cerebellum]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.12.22.473884v1?rss=1">
<title>
<![CDATA[
Study of an FBXO7 patient mutation reveals Fbxo7 and PI31 co-regulate proteasomes and mitochondria 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.12.22.473884v1?rss=1"
</link>
<description><![CDATA[
Mutations in FBXO7 have been discovered associated with an atypical parkinsonism. We report here a new homozygous missense mutation in a paediatric patient that causes an L250P substitution in the dimerization domain of Fbxo7. This alteration selectively ablates the Fbxo7-PI31 interaction and causes a significant reduction in Fbxo7 and PI31 levels in patient cells. Consistent with their association with proteasomes, L250P patient fibroblasts have reduced proteasome activity and proteasome subunits. We also show PI31 interacts directly with the MiD49/51 fission adaptor proteins, and unexpectedly, PI31 acts as an adaptor enabling SCFFbxo7 ligase to ubiquitinate MiD49. Thus, the L250P mutation changes the function of Fbxo7 by altering its substrate repertoire. Although MiD49/51 expression was reduced in L250P patient cells, there was no effect on the mitochondrial network. However, patient cells had higher levels of ROS and reduced viability under stress. Our study shows that Fbxo7 and PI31 affect each others functions in regulating both proteasomal and mitochondrial function and demonstrate a new function for PI31, as an adaptor for the SCFFbxo7 E3 ubiquitin ligase.
]]></description>
<dc:creator>Al Rawi, S.</dc:creator>
<dc:creator>Simpson, L.</dc:creator>
<dc:creator>McDonald, N. Q.</dc:creator>
<dc:creator>Chernuha, V.</dc:creator>
<dc:creator>Elpeleg, O.</dc:creator>
<dc:creator>Zeviani, M. Q.</dc:creator>
<dc:creator>Barker, R. A.</dc:creator>
<dc:creator>Spiegel, R.</dc:creator>
<dc:creator>Laman, H.</dc:creator>
<dc:date>2021-12-23</dc:date>
<dc:identifier>doi:10.1101/2021.12.22.473884</dc:identifier>
<dc:title><![CDATA[Study of an FBXO7 patient mutation reveals Fbxo7 and PI31 co-regulate proteasomes and mitochondria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.12.22.473955v1?rss=1">
<title>
<![CDATA[
Regulation of KHNYN antiviral activity by the extended di-KH domain and nucleo-cytoplasmic trafficking 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.12.22.473955v1?rss=1"
</link>
<description><![CDATA[
The zinc finger antiviral protein (ZAP) restricts a broad range of viruses by binding CpG dinucleotides in viral RNA to target it for degradation and inhibit its translation. KHNYN was recently identified as an antiviral protein required for ZAP to inhibit retroviral replication, though little is known about its functional determinants. KHNYN contains an N-terminal extended di-KH-like domain, a PIN endoribonuclease domain and a C-terminal CUBAN domain that binds NEDD8 and ubiquitin. We show that deletion of the extended di-KH domain reduces its antiviral activity. However, despite its similarity to RNA binding KH domains, the extended di-KH domain in KHNYN does not appear to bind RNA. Mutation of residues in the CUBAN domain that bind NEDD8 increase KHNYN abundance but do not alter its antiviral activity, suggesting that this interaction regulates KHNYN homeostatic turnover. In contrast, a CRM1-dependent nuclear export signal (NES) at the C-terminus of the CUBAN domain is required for antiviral activity. Deletion of this signal retains KHNYN in the nucleus and inhibits its interaction with ZAP. Interestingly, this NES appeared in the KHNYN lineage at a similar time as when ZAP evolved in tetrapods, indicating that these proteins may have co-evolved to restrict viral replication.

AUTHOR SUMMARYAntiviral proteins restrict viral replication in many different ways, including inhibiting viral gene expression. ZAP is an antiviral RNA binding protein that must interact with other cellular proteins to inhibit viral protein synthesis. KHNYN is a ZAP cofactor that is required for it to inhibit retroviral replication. Because little is known about how KHNYN functions in this role, we have analyzed how two of its domains regulate its antiviral activity. We first show that the extended di-KH domain in KHNYN is required for its antiviral activity. While it is related to di-KH domains in RNA binding proteins, it appears to have lost its ability to bind RNA and KHNYN likely acts in the restriction pathway after ZAP binds a target viral RNA. Second, we show that the KHNYN CUBAN domain regulates both its protein abundance and trafficking within the cell. The CUBAN domain contains a nuclear export signal and, when this signal is mutated, KHNYN is sequestered in the nucleus, has substantially reduced antiviral activity and does not interact with ZAP. Overall, we show that the extended di-KH and CUBAN domains in KHNYN are required for it to act as a cofactor for ZAP to restrict viral replication.
]]></description>
<dc:creator>Lista, M. J.</dc:creator>
<dc:creator>Galao, R. P.</dc:creator>
<dc:creator>Ficarelli, M.</dc:creator>
<dc:creator>Kmiec, D.</dc:creator>
<dc:creator>Wilson, H.</dc:creator>
<dc:creator>Winstone, H.</dc:creator>
<dc:creator>Morris, E. R.</dc:creator>
<dc:creator>Mischo, H. E.</dc:creator>
<dc:creator>Wanford, J.</dc:creator>
<dc:creator>Youle, R.</dc:creator>
<dc:creator>Odendall, C.</dc:creator>
<dc:creator>Taylor, I. A.</dc:creator>
<dc:creator>Neil, S. J.</dc:creator>
<dc:creator>Swanson, C.</dc:creator>
<dc:date>2021-12-24</dc:date>
<dc:identifier>doi:10.1101/2021.12.22.473955</dc:identifier>
<dc:title><![CDATA[Regulation of KHNYN antiviral activity by the extended di-KH domain and nucleo-cytoplasmic trafficking]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.12.30.474570v1?rss=1">
<title>
<![CDATA[
Endosomal structure and APP biology are not altered in preclinical cellular models of Down syndrome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.12.30.474570v1?rss=1"
</link>
<description><![CDATA[
Individuals who have Down syndrome (trisomy 21) are at greatly increased risk of developing Alzheimers disease - dementia. Alzheimers disease is characterised by the accumulation in the brain of amyloid-{beta} plaques that are a product of amyloid precursor protein, encoded by the APP gene on chromosome 21. In Down syndrome the first site of amyloid-{beta} accumulation is within endosomes and changes to endosome biology occur early in disease. Here we determine if primary mouse embryonic fibroblasts isolated from two mouse models of Down syndrome can be used to study endosome and APP cell biology. We report that in these cellular models of Down syndrome endosome number, size and APP processing are not altered, likely because APP is not dosage sensitive in these models, despite three copies of App.
]]></description>
<dc:creator>Cannavo, C.</dc:creator>
<dc:creator>Cleverley, K.</dc:creator>
<dc:creator>Maduro, C.</dc:creator>
<dc:creator>Mumford, P.</dc:creator>
<dc:creator>Moulding, D.</dc:creator>
<dc:creator>Fisher, E.</dc:creator>
<dc:creator>Wiseman, F. K.</dc:creator>
<dc:date>2021-12-30</dc:date>
<dc:identifier>doi:10.1101/2021.12.30.474570</dc:identifier>
<dc:title><![CDATA[Endosomal structure and APP biology are not altered in preclinical cellular models of Down syndrome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.12.16.472890v1?rss=1">
<title>
<![CDATA[
An RNA-binding switch drives ribosome biogenesis and tumorigenesis downstream of RAS oncogene 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.12.16.472890v1?rss=1"
</link>
<description><![CDATA[
Oncogenic RAS signaling reprograms gene expression through both transcriptional and post-transcriptional mechanisms. While transcriptional regulation downstream of RAS is relatively well characterized, how RAS post-transcriptionally modulates gene expression to promote malignancy is largely unclear. Using quantitative RNA Interactome Capture analysis, we reveal that oncogenic RAS signaling reshapes the RNA-bound proteomic landscape of cancer cells, with a network of nuclear proteins centered around Nucleolin displaying enhanced RNA-binding activity. We show that Nucleolin is phosphorylated downstream of RAS, which increases its binding to pre-ribosomal-RNA (rRNA), boosts rRNA production, and promotes ribosome biogenesis. This Nucleolin-dependent enhancement of ribosome biogenesis is crucial for RAS-induced cancer cell proliferation, and can be targeted therapeutically to inhibit tumor growth. Our results reveal that oncogenic RAS signaling drives ribosome biogenesis by regulating the RNA-binding activity of Nucleolin, and highlight the crucial role of this process in RAS-mediated tumorigenesis.
]]></description>
<dc:creator>Azman, M. S.</dc:creator>
<dc:creator>Dodel, M.</dc:creator>
<dc:creator>Capraro, F.</dc:creator>
<dc:creator>Faraway, R.</dc:creator>
<dc:creator>Dermit, M.</dc:creator>
<dc:creator>Fan, W.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Mardakheh, F. K.</dc:creator>
<dc:date>2021-12-16</dc:date>
<dc:identifier>doi:10.1101/2021.12.16.472890</dc:identifier>
<dc:title><![CDATA[An RNA-binding switch drives ribosome biogenesis and tumorigenesis downstream of RAS oncogene]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.01.04.474788v1?rss=1">
<title>
<![CDATA[
Unraveling membrane properties at the organelle-level with LipidDyn 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.01.04.474788v1?rss=1"
</link>
<description><![CDATA[
Cellular membranes are formed from many different lipids in various amounts and proportions depending on the subcellular localization. The lipid composition of membranes is sensitive to changes in the cellular environment, and their alterations are linked to several diseases, including cancer. Lipids not only form lipid-lipid interactions but also interact with other biomolecules, including proteins, profoundly impacting each other.

Molecular dynamics (MD) simulations are a powerful tool to study the properties of cellular membranes and membrane-protein interactions on different timescales and at varying levels of resolution. Over the last few years, software and hardware for biomolecular simulations have been optimized to routinely run long simulations of large and complex biological systems. On the other hand, high-throughput techniques based on lipidomics provide accurate estimates of the composition of cellular membranes at the level of subcellular compartments. The community needs computational tools for lipidomics and simulation data effectively interacting to better understand how changes in lipid compositions impact membrane function and structure. Lipidomic data can be analyzed to design biologically relevant models of membranes for MD simulations. Similar applications easily result in a massive amount of simulation data where the bottleneck becomes the analysis of the data to understand how membrane properties and membrane-protein interactions are changing in the different conditions. In this context, we developed LipidDyn, an in silico pipeline to streamline the analyses of MD simulations of membranes of different compositions. Once the simulations are collected, LipidDyn provides average properties and time series for several membrane properties such as area per lipid, thickness, diffusion motions, the density of lipid bilayers, and lipid enrichment/depletion. The calculations exploit parallelization and the pipelines include graphical outputs in a publication-ready form. We applied LipidDyn to different case studies to illustrate its potential, including membranes from cellular compartments and transmembrane protein domains. LipidDyn is implemented in Python and relies on open-source libraries. LipidDyn is available free of charge under the GNU General Public License from https://github.com/ELELAB/LipidDyn.
]]></description>
<dc:creator>Scrima, S.</dc:creator>
<dc:creator>Tiberti, M.</dc:creator>
<dc:creator>Campo, A.</dc:creator>
<dc:creator>Corcelle-Termeau, E.</dc:creator>
<dc:creator>Judith, D.</dc:creator>
<dc:creator>Moller Foged, M.</dc:creator>
<dc:creator>Bundgaard Clemmens, K. K.</dc:creator>
<dc:creator>Tooze, S.</dc:creator>
<dc:creator>Jäättelä, M.</dc:creator>
<dc:creator>Maeda, K.</dc:creator>
<dc:creator>Lambrughi, M.</dc:creator>
<dc:creator>Papaleo, E.</dc:creator>
<dc:date>2022-01-05</dc:date>
<dc:identifier>doi:10.1101/2022.01.04.474788</dc:identifier>
<dc:title><![CDATA[Unraveling membrane properties at the organelle-level with LipidDyn]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.12.14.472728v1?rss=1">
<title>
<![CDATA[
Lactate regulation of activation in CD8+ T cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.12.14.472728v1?rss=1"
</link>
<description><![CDATA[
CD8+ T cells infiltrate virtually every tissue to find and destroy infected or mutated cells. They often traverse varying oxygen levels and nutrient-deprived microenvironments. High glycolytic activity in tissues can result in extended exposure of cytotoxic T cells to the metabolite lactate. Lactate can be immunosuppressive, at least in part due to its association with tissue acidosis. We show here that the lactate anion is well tolerated by CD8+ T cells in pH neutral conditions. We describe how lactate is taken up by activated CD8+ T cells and is capable of displacing glucose as a carbon source. Activation in the presence of a pH neutral form of lactate significantly alters the CD8+ T cell transcriptome, including the expression of key effector differentiation markers such as granzyme B and interferon-gamma. Our studies reveal the novel metabolic features of lactate utilization by activated CD8+ T cells, and highlight the importance of lactate in shaping the differentiation and activity of cytotoxic T cells.
]]></description>
<dc:creator>Barbieri, L.</dc:creator>
<dc:creator>Velica, P.</dc:creator>
<dc:creator>Gameiro, P. A.</dc:creator>
<dc:creator>Cunha, P. P.</dc:creator>
<dc:creator>Foskolou, I. P.</dc:creator>
<dc:creator>Bargiela, D.</dc:creator>
<dc:creator>Rundqvist, H.</dc:creator>
<dc:creator>Johnson, R.</dc:creator>
<dc:date>2021-12-17</dc:date>
<dc:identifier>doi:10.1101/2021.12.14.472728</dc:identifier>
<dc:title><![CDATA[Lactate regulation of activation in CD8+ T cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.01.09.475567v1?rss=1">
<title>
<![CDATA[
Interactions between influenza A virus nucleoprotein and gene segment UTRs facilitate selective modulation of viral gene expression 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.01.09.475567v1?rss=1"
</link>
<description><![CDATA[
The influenza A virus (IAV) genome is divided into eight negative-sense, single-stranded RNA segments. Each segment exhibits a unique level and temporal pattern of expression, however the exact mechanisms underlying the patterns of individual gene segment expression are poorly understood. We previously demonstrated that a single substitution in the viral nucleoprotein (NP:F346S) selectively modulates neuraminidase (NA) gene segment expression while leaving other segments largely unaffected. Given what is currently known about NP function, there is no obvious explanation for how changes in NP can selectively modulate the replication of individual gene segments. We found that the specificity of this effect for the NA segment is virus strain specific and depends on the UTR sequences of the NA segment. While the NP:F346S substitution did not significantly alter the RNA binding or oligomerization activities of NP in vitro, it specifically decreased the ability of NP to promote NA segment vRNA synthesis. In addition to NP residue F346, we identified two other adjacent aromatic residues in NP (Y385 & F479) capable of similarly regulating NA gene segment expression, suggesting a larger role for this domain in gene-segment specific regulation. Our findings reveal a new role for NP in selective regulation of viral gene segment replication and demonstrate how the expression patterns of individual viral gene segments can be modulated during adaptation to new host environments.

Author summaryInfluenza A virus (IAV) is a respiratory pathogen that remains a significant source of morbidity and mortality. Escape from host immunity or emergence into new host species often requires mutations that modulate the functional activities of the viral glycoproteins hemagglutinin (HA) and neuraminidase (NA) which are responsible for virus attachment to and release from host cells, respectively. Maintaining the functional balance between the activities of HA and NA is required for fitness across multiple host systems. Thus, selective modulation of viral gene expression patterns may be a key determinant of viral immune escape and cross-species transmission potential. We identified a novel mechanism by which the viral nucleoprotein (NP) gene can selectively modulate NA segment replication and gene expression through interactions with the segment UTR. Our work highlights an unexpected role for NP in selective regulation of expression from the individual IAV gene segments.
]]></description>
<dc:creator>Diefenbacher, M.</dc:creator>
<dc:creator>Tan, T. J. C.</dc:creator>
<dc:creator>Bauer, D. L. V.</dc:creator>
<dc:creator>Stadtmueller, B.</dc:creator>
<dc:creator>Wu, N. C.</dc:creator>
<dc:creator>brooke, c.</dc:creator>
<dc:date>2022-01-09</dc:date>
<dc:identifier>doi:10.1101/2022.01.09.475567</dc:identifier>
<dc:title><![CDATA[Interactions between influenza A virus nucleoprotein and gene segment UTRs facilitate selective modulation of viral gene expression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.01.12.475985v1?rss=1">
<title>
<![CDATA[
Role of Yap1 in adult neural stem cell activation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.01.12.475985v1?rss=1"
</link>
<description><![CDATA[
Most adult hippocampal neural stem cells (NSCs) remain quiescent with only a minor portion undergoing active proliferation and neurogenesis. The molecular mechanisms that trigger eventually the transition from quiescence to activation are still poorly understood. Here, we found the activity of the transcriptional activator Yap1 to be enriched in active NSCs. Genetic deletion of Yap1 led to a significant reduction in the relative proportion of active NSCs supporting a physiological role of Yap1 in regulating the transition from quiescence to activation. Overexpression of wild type Yap1 in adult NSCs did not induce NSC activation suggesting tight upstream control mechanisms, but overexpression of a gain-of-function mutant (Yap1-5SA) elicited cell cycle entry in NSCs and hilar astrocytes. Consistent with a role of Yap1 in NSC activation, single cell RNA sequencing revealed the partial induction of an activated NSC gene expression program. Yet, Yap1-5SA expression also induced Taz and other key components of the Yap/Taz regulon previously identified in glioblastoma stem cell-like cells. Consequently, dysregulated Yap1 activity led to repression of hippocampal neurogenesis, promoting aberrant differentiation instead.
]]></description>
<dc:creator>Berninger, B.</dc:creator>
<dc:creator>Fan, W.</dc:creator>
<dc:creator>Jurado-Arjona, J.</dc:creator>
<dc:creator>Alanis-Lobato, G.</dc:creator>
<dc:creator>Peron, S.</dc:creator>
<dc:creator>Berger, C.</dc:creator>
<dc:creator>Andrade-Navarro, M. A.</dc:creator>
<dc:creator>Falk, S.</dc:creator>
<dc:date>2022-01-13</dc:date>
<dc:identifier>doi:10.1101/2022.01.12.475985</dc:identifier>
<dc:title><![CDATA[Role of Yap1 in adult neural stem cell activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.01.14.476382v1?rss=1">
<title>
<![CDATA[
Selection analysis identifies unusual clustered mutational changes in Omicron lineage BA.1 that likely impact Spike function 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.01.14.476382v1?rss=1"
</link>
<description><![CDATA[
Among the 30 non-synonymous nucleotide substitutions in the Omicron S-gene are 13 that have only rarely been seen in other SARS-CoV-2 sequences. These mutations cluster within three functionally important regions of the S-gene at sites that will likely impact (i) interactions between subunits of the Spike trimer and the predisposition of subunits to shift from down to up configurations, (ii) interactions of Spike with ACE2 receptors, and (iii) the priming of Spike for membrane fusion. We show here that, based on both the rarity of these 13 mutations in intrapatient sequencing reads and patterns of selection at the codon sites where the mutations occur in SARS-CoV-2 and related sarbecoviruses, prior to the emergence of Omicron the mutations would have been predicted to decrease the fitness of any genomes within which they occurred. We further propose that the mutations in each of the three clusters therefore cooperatively interact to both mitigate their individual fitness costs, and adaptively alter the function of Spike. Given the evident epidemic growth advantages of Omicron over all previously known SARS-CoV-2 lineages, it is crucial to determine both how such complex and highly adaptive mutation constellations were assembled within the Omicron S-gene, and why, despite unprecedented global genomic surveillance efforts, the early stages of this assembly process went completely undetected.
]]></description>
<dc:creator>Martin, D. P.</dc:creator>
<dc:creator>Lytras, S.</dc:creator>
<dc:creator>Lucaci, A. G.</dc:creator>
<dc:creator>Maier, W.</dc:creator>
<dc:creator>Gruning, B.</dc:creator>
<dc:creator>Shank, S. D.</dc:creator>
<dc:creator>Weaver, S.</dc:creator>
<dc:creator>MacLean, O. S.</dc:creator>
<dc:creator>Orton, R. J.</dc:creator>
<dc:creator>Lemey, P.</dc:creator>
<dc:creator>Boni, M. F.</dc:creator>
<dc:creator>Tegally, H.</dc:creator>
<dc:creator>Harkins, G. W.</dc:creator>
<dc:creator>Scheepers, C.</dc:creator>
<dc:creator>Bhiman, J. N.</dc:creator>
<dc:creator>Everatt, J.</dc:creator>
<dc:creator>Amoako, D. G.</dc:creator>
<dc:creator>San, J. E.</dc:creator>
<dc:creator>Giandhari, J.</dc:creator>
<dc:creator>Sigal, A.</dc:creator>
<dc:creator>NGS-SA,</dc:creator>
<dc:creator>Williamson, C.</dc:creator>
<dc:creator>Hsiao, N.-y.</dc:creator>
<dc:creator>von Gottberg, A.</dc:creator>
<dc:creator>De Klerk, A.</dc:creator>
<dc:creator>Shafer, R. W.</dc:creator>
<dc:creator>Robertson, D. L.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Sewell, B. T.</dc:creator>
<dc:creator>Lessells, R.</dc:creator>
<dc:creator>Nekrutenko, A.</dc:creator>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Murrell, B.</dc:creator>
<dc:creator>Wilkinson, E.</dc:creator>
<dc:creator>Gupta, R. K.</dc:creator>
<dc:creator>de Oliveira, T.</dc:creator>
<dc:creator>Kosakovsky Pond, S. L.</dc:creator>
<dc:date>2022-01-18</dc:date>
<dc:identifier>doi:10.1101/2022.01.14.476382</dc:identifier>
<dc:title><![CDATA[Selection analysis identifies unusual clustered mutational changes in Omicron lineage BA.1 that likely impact Spike function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.01.18.476868v1?rss=1">
<title>
<![CDATA[
CDC50 orthologues in Plasmodium falciparum have distinct roles in merozoite egress and trophozoite maturation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.01.18.476868v1?rss=1"
</link>
<description><![CDATA[
In model organisms P4-ATPases require cell division control protein 50 (CDC50) chaperones for their phospholipid flipping activity. In the malaria parasite, P. falciparum, guanylyl cyclase alpha (GC) is an integral membrane protein that is essential for release (egress) of merozoites from their host erythrocytes. GC is unusual in that it contains both a C-terminal cyclase domain and an N-terminal P4-ATPase domain of unknown function. We sought to investigate whether any of the three CDC50 orthologues (denoted A, B and C) encoded by P. falciparum are required for GC function. Using gene tagging and conditional gene disruption, we demonstrate that both CDC50B and CDC50C are expressed in the clinically important asexual blood stages and that CDC50B is a binding partner of GC whereas CDC50C is the binding partner of another putative P4-ATPase, ATP2. Our findings indicate that CDC50B has no essential role for intraerythrocytic parasite maturation but modulates the rate of parasite egress by interacting with GC for optimal cGMP synthesis. In contrast, CDC50C is essential for blood stage trophozoite maturation. Additionally, we find that the CDC50C-ATP2 complex may influence parasite endocytosis of host cell haemoglobin and consequently hemozoin formation.
]]></description>
<dc:creator>Patel, A.</dc:creator>
<dc:creator>Nofal, S. D.</dc:creator>
<dc:creator>Blackman, M.</dc:creator>
<dc:creator>Baker, D. A.</dc:creator>
<dc:date>2022-01-20</dc:date>
<dc:identifier>doi:10.1101/2022.01.18.476868</dc:identifier>
<dc:title><![CDATA[CDC50 orthologues in Plasmodium falciparum have distinct roles in merozoite egress and trophozoite maturation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.01.17.476508v1?rss=1">
<title>
<![CDATA[
Machine learning guided signal enrichment for ultrasensitive plasma tumor burden monitoring 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.01.17.476508v1?rss=1"
</link>
<description><![CDATA[
In solid tumor oncology, circulating tumor DNA (ctDNA) is poised to transform care through accurate assessment of minimal residual disease (MRD) and therapeutic response monitoring. To overcome the sparsity of ctDNA fragments in low tumor fraction (TF) settings and increase MRD sensitivity, we previously leveraged genome-wide mutational integration through plasma whole genome sequencing (WGS). We now introduce MRD-EDGE, a composite machine learning-guided WGS ctDNA single nucleotide variant (SNV) and copy number variant (CNV) detection platform designed to increase signal enrichment. MRD-EDGE uses deep learning and a ctDNA-specific feature space to increase SNV signal to noise enrichment in WGS by 300X compared to our previous noise suppression platform MRDetect. MRD-EDGE also reduces the degree of aneuploidy needed for ultrasensitive CNV detection through WGS from 1Gb to 200Mb, thereby expanding its applicability to a wider range of solid tumors. We harness the improved performance to track changes in tumor burden in response to neoadjuvant immunotherapy in non-small cell lung cancer and demonstrate ctDNA shedding in precancerous colorectal adenomas. Finally, the radical signal to noise enrichment in MRD-EDGE enables de novo mutation calling in melanoma without matched tumor, yielding clinically informative TF monitoring for patients on immune checkpoint inhibition.
]]></description>
<dc:creator>Widman, A. J.</dc:creator>
<dc:creator>Shah, M.</dc:creator>
<dc:creator>Ogaard, N.</dc:creator>
<dc:creator>Khamnei, C. C.</dc:creator>
<dc:creator>Frydendahl, A.</dc:creator>
<dc:creator>Deshpande, A.</dc:creator>
<dc:creator>Arora, A.</dc:creator>
<dc:creator>Zhang, M.</dc:creator>
<dc:creator>Halmos, D.</dc:creator>
<dc:creator>Bass, J.</dc:creator>
<dc:creator>Langanay, T.</dc:creator>
<dc:creator>Rajagopalan, S.</dc:creator>
<dc:creator>Steinsnyder, Z.</dc:creator>
<dc:creator>Liao, W.</dc:creator>
<dc:creator>Rasmussen, M. H.</dc:creator>
<dc:creator>Jensen, S. O.</dc:creator>
<dc:creator>Nors, J.</dc:creator>
<dc:creator>Therkildsen, C.</dc:creator>
<dc:creator>Sotelo, J.</dc:creator>
<dc:creator>Brand, R.</dc:creator>
<dc:creator>Shah, R. H.</dc:creator>
<dc:creator>Cheng, A. P.</dc:creator>
<dc:creator>Maher, C.</dc:creator>
<dc:creator>Spain, L.</dc:creator>
<dc:creator>Krause, K.</dc:creator>
<dc:creator>Frederick, D. T.</dc:creator>
<dc:creator>Malbari, M. S.</dc:creator>
<dc:creator>Marton, M.</dc:creator>
<dc:creator>Manaa, D.</dc:creator>
<dc:creator>Winterkorn, L.</dc:creator>
<dc:creator>Callahan, M. K.</dc:creator>
<dc:creator>Boland, G.</dc:creator>
<dc:creator>Wolchok, J. D.</dc:creator>
<dc:creator>Saxena, A.</dc:creator>
<dc:creator>Turajlic, S.</dc:creator>
<dc:creator>Imielinski, M.</dc:creator>
<dc:creator>Berger, M. F.</dc:creator>
<dc:creator>Altorki, N. K.</dc:creator>
<dc:creator>Postow, M. A.</dc:creator>
<dc:creator>Robine, N.</dc:creator>
<dc:creator>Andersen, C. L.</dc:creator>
<dc:creator>Landau, D. A.</dc:creator>
<dc:date>2022-01-20</dc:date>
<dc:identifier>doi:10.1101/2022.01.17.476508</dc:identifier>
<dc:title><![CDATA[Machine learning guided signal enrichment for ultrasensitive plasma tumor burden monitoring]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.01.19.476881v1?rss=1">
<title>
<![CDATA[
SAIBR: A simple, platform-independent method for spectral autofluorescence correction 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.01.19.476881v1?rss=1"
</link>
<description><![CDATA[
Biological systems are increasingly viewed through the lens of mathematics, physics, and systems approaches that demand accurate quantification of gene expression and local protein concentrations. Such approaches have benefited greatly from the revolution in genetic engineering sparked by CRISPR/Cas9. By facilitating the tagging of genes at their genomic loci, CRISPR/Cas9 allows us to use fluorescence to monitor proteins that are expressed at or near endogenous levels under native regulatory control. However, due to their typically lower expression levels, quantitative experiments using endogenously-tagged genes can run into limits imposed by autofluorescence (AF). AF is often a particular challenge in the illumination bands occupied by the most efficient fluorescent proteins (GFP, mNeonGreen). Stimulated by our work in C. elegans, we describe and validate Spectral Autofluorescence Image correction By Regression (SAIBR), a simple, platform-independent protocol, and associated GUI-based FIJI plugin to correct for autofluorescence using standard filter sets and illumination conditions. Fully validated for use in C. elegans embryos and tested in diverse systems, including starfish oocytes and fission yeast, SAIBR achieves accurate quantitation of fluorophore signal and enables reliable detection and quantification of even weakly expressed proteins. Thus, SAIBR provides a highly accessible, low barrier way to incorporate AF correction as standard for researchers working on a broad variety of cell and developmental systems.

Summary StatementImplemented as an easy-to-use Fiji Plugin, SAIBR provides effective autofluorescence correction for cells and tissues using standard imaging conditions.
]]></description>
<dc:creator>Rodrigues, N. T. L.</dc:creator>
<dc:creator>Bland, T.</dc:creator>
<dc:creator>Borrego-Pinto, J.</dc:creator>
<dc:creator>Ng, K.</dc:creator>
<dc:creator>Hirani, N.</dc:creator>
<dc:creator>Gu, Y.</dc:creator>
<dc:creator>Foo, S.</dc:creator>
<dc:creator>Goehring, N. W.</dc:creator>
<dc:date>2022-01-20</dc:date>
<dc:identifier>doi:10.1101/2022.01.19.476881</dc:identifier>
<dc:title><![CDATA[SAIBR: A simple, platform-independent method for spectral autofluorescence correction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.01.20.477035v1?rss=1">
<title>
<![CDATA[
m6A reader Pho92 is recruited co-transcriptionally and couples translation efficacy to mRNA decay to promote meiotic fitness in yeast 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.01.20.477035v1?rss=1"
</link>
<description><![CDATA[
N6-methyladenosine (m6A) RNA modification impacts mRNA fate primarily via reader proteins, which dictate processes in development, stress, and disease. Yet little is known about m6A function in Saccharomyces cerevisiae, which occurs solely during early meiosis. Here we perform a multifaceted analysis of the m6A reader protein Pho92/Mrb1. Cross-linking immunoprecipitation analysis reveals that Pho92 associates with the 3end of meiotic mRNAs in both an m6A-dependent and independent manner. Within cells, Pho92 transitions from the nucleus to the cytoplasm, and associates with translating ribosomes. In the nucleus Pho92 associates with target loci through its interaction with transcriptional elongator Paf1C. Functionally, we show that Pho92 promotes and links protein synthesis to mRNA decay. As such, the Pho92-mediated m6A-mRNA decay is contingent on active translation and the CCR4-NOT complex. We propose that the m6A reader Pho92 is loaded co-transcriptionally to facilitate protein synthesis and subsequent decay of m6A modified transcripts, and thereby promotes meiosis.
]]></description>
<dc:creator>Varier, R. A.</dc:creator>
<dc:creator>Sideri, T.</dc:creator>
<dc:creator>Capitanchik, C.</dc:creator>
<dc:creator>Manova, Z.</dc:creator>
<dc:creator>Calvani, E.</dc:creator>
<dc:creator>Rossi, A.</dc:creator>
<dc:creator>Edupuganti, R.</dc:creator>
<dc:creator>Ensinck, I.</dc:creator>
<dc:creator>Chan, V. W. C.</dc:creator>
<dc:creator>Patel, H.</dc:creator>
<dc:creator>Kirkpatrick, J.</dc:creator>
<dc:creator>Faull, P.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Vermeulen, M.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:creator>van Werven, F.</dc:creator>
<dc:date>2022-01-21</dc:date>
<dc:identifier>doi:10.1101/2022.01.20.477035</dc:identifier>
<dc:title><![CDATA[m6A reader Pho92 is recruited co-transcriptionally and couples translation efficacy to mRNA decay to promote meiotic fitness in yeast]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.01.19.476598v1?rss=1">
<title>
<![CDATA[
Tissue-intrinsic γδ T cells critically regulate Tissue-Resident Memory CD8 T cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.01.19.476598v1?rss=1"
</link>
<description><![CDATA[
Because Tissue-Resident Memory T (TRM) cells contribute critically to body-surface immunoprotection and/or immunopathology in multiple settings, their regulation is biologically and clinically important. Interestingly, TRM commonly develop in epithelia part-shaped by innate-like lymphocytes that become tissue-intrinsic during development. Here we show that polyclonal TRM cells induced by allergic contact dermatitis (ACD) interact with signature intraepidermal {gamma}{delta} T cells, facilitating a feedback-loop wherein TRM-derived IFN{gamma} upregulates PD-L1 on {gamma}{delta} cells that can thereupon regulate PD1+ TRM. Thus, TRM induced by ACD in mice lacking either local {gamma}{delta} cells, or lacking a single gene (IFN{gamma}R) expressed by local {gamma}{delta} cells, displayed enhanced proliferative and effector potentials. Those phenotypes were associated with strikingly limited motility, reduced TRM quality. and an impaired capacity to restrain melanoma. Thus, inter-individual and tissue-specific variation in how tissue-intrinsic lymphocytes integrate with TRM may sit upstream of variation in responses to cancer, allergens and other challenges, and may likewise underpin inflammatory pathologies repeatedly observed in {gamma}{delta}-deficient animals.
]]></description>
<dc:creator>Munoz-Ruiz, M.</dc:creator>
<dc:creator>Llorian, M.</dc:creator>
<dc:creator>D'Antuono, R.</dc:creator>
<dc:creator>Pavlova, A.</dc:creator>
<dc:creator>Mavrigiannaki, A. M.</dc:creator>
<dc:creator>Mckenzie, D.</dc:creator>
<dc:creator>Garcia-Cassani, B.</dc:creator>
<dc:creator>Iannitto, M. L.</dc:creator>
<dc:creator>Jandke, A.</dc:creator>
<dc:creator>Ushakov, D.</dc:creator>
<dc:creator>Adrian C. Hayday, A.</dc:creator>
<dc:date>2022-01-21</dc:date>
<dc:identifier>doi:10.1101/2022.01.19.476598</dc:identifier>
<dc:title><![CDATA[Tissue-intrinsic γδ T cells critically regulate Tissue-Resident Memory CD8 T cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.01.25.477676v1?rss=1">
<title>
<![CDATA[
The rates of adult neurogenesis and oligodendrogenesis are linked to cell cycle regulation through p27-dependent gene repression of SOX2 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.01.25.477676v1?rss=1"
</link>
<description><![CDATA[
Cell differentiation involves profound changes in global gene expression that often have to occur in coordination with cell cycle exit. Because cyclin-dependent kinase inhibitor p27 reportedly regulates proliferation of neural progenitor cells in the subependymal neurogenic niche of the adult mouse brain, but can also have effects on gene expression, we decided to molecularly analyze its role in adult neurogenesis and oligodendrogenesis. At the cell level, we show that p27 restricts residual cyclin-dependent kinase activity after mitogen withdrawal to antagonize cycling, but it is not essential for cell cycle exit. Contrasting gene expression with chromatin accessibility, we find that p27 is coincidentally necessary to globally repress many genes involved in the transit from multipotentiality to differentiation, including those coding for neural progenitor transcription factors SOX2, OLIG2, and ASCL1. Our data reveal both direct association of p27 with regulatory sequences in the three genes and an additional hierarchical relationship where p27 repression of the Sox2 gene leads to reduced levels of SOX2-downstream targets Olig2 and Ascl1. In vivo, p27 is also required for the regulation of the proper level of SOX2 necessary for neuroblasts and oligodendroglial progenitor cells to timely exit cell cycle in a lineage-dependent manner.
]]></description>
<dc:creator>Domingo-Muelas, A.</dc:creator>
<dc:creator>Morante-Redolat, J. M.</dc:creator>
<dc:creator>Moncho-Amor, V.</dc:creator>
<dc:creator>Jordan, A.</dc:creator>
<dc:creator>Perez-Villalba, A.</dc:creator>
<dc:creator>Carrillo-Barbera, P.</dc:creator>
<dc:creator>Belenguer, G.</dc:creator>
<dc:creator>Porlan, E.</dc:creator>
<dc:creator>Kirstein, M.</dc:creator>
<dc:creator>Bachs, O.</dc:creator>
<dc:creator>Ferron, S. R.</dc:creator>
<dc:creator>Lovell-Badge, R.</dc:creator>
<dc:creator>Farinas, I.</dc:creator>
<dc:date>2022-01-25</dc:date>
<dc:identifier>doi:10.1101/2022.01.25.477676</dc:identifier>
<dc:title><![CDATA[The rates of adult neurogenesis and oligodendrogenesis are linked to cell cycle regulation through p27-dependent gene repression of SOX2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.01.25.477674v1?rss=1">
<title>
<![CDATA[
ARPC5 Isoforms Drive Distinct Arp2/3-dependant Actin Remodeling Events in CD4 T Cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.01.25.477674v1?rss=1"
</link>
<description><![CDATA[
Arp2/3-dependent formation of nuclear F-actin networks of different morphology and stability is observed in an increasing number of biological processes. In CD4 T cells, T cell receptor (TCR) signaling induces cytoplasmic and nuclear F-actin assembly via Arp2/3 to strengthen contacts to antigen presenting cells and to regulate gene expression, respectively. How Arp2/3 complex is regulated to mediate these distinct actin polymerization events in response to a common stimulus is unknown. Arp2/3-complex consists of 7 subunits where ARP3, ARPC1 and ARPC5 exist as two different isoforms in humans that can assemble in complexes with different properties. Examining whether specific Arp2/3 subunit isoforms govern distinct actin remodeling events in CD4 T cells, we find that the ARPC5L isoform drives nuclear actin polymerization, while cytoplasmic actin dynamics and TCR proximal signalling selectively relies on ARPC5. In contrast, formation of stable nuclear F-actin networks triggered by DNA replication stress in CD4 T cells requires ARPC5 and is independent of ARPC5L. Moreover, nuclear actin polymerization induced by TCR signaling but not by DNA replication stress is controlled by nuclear calcium-calmodulin signalling and N-WASP. Specific ARPC5 isoforms thus govern Arp2/3 complex activity in distinct actin polymerization events. ARPC5 isoform diversity thus emerges as a mechanism to tailor Arp2/3 activity to different physiological stimuli.
]]></description>
<dc:creator>Sadhu, L.</dc:creator>
<dc:creator>Tsopoulidis, N.</dc:creator>
<dc:creator>Laketa, V.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:creator>Fackler, O.</dc:creator>
<dc:date>2022-01-26</dc:date>
<dc:identifier>doi:10.1101/2022.01.25.477674</dc:identifier>
<dc:title><![CDATA[ARPC5 Isoforms Drive Distinct Arp2/3-dependant Actin Remodeling Events in CD4 T Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.01.25.476955v1?rss=1">
<title>
<![CDATA[
Transient RNA structures cause aberrant influenza virus replication and innate immune activation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.01.25.476955v1?rss=1"
</link>
<description><![CDATA[
During infection, the influenza A virus RNA polymerase produces both full-length and aberrant RNA molecules, such as defective viral genomes (DVG) and mini viral RNAs (mvRNA). Subsequent innate immune activation involves the binding of host pathogen receptor retinoic acid-inducible gene I (RIG-I) to viral RNAs. However, not all influenza A virus RNAs are strong RIG-I agonists. Here we show that potent innate immune activation by mvRNAs is determined by transient RNA structures, called template loops (t-loop) that stall the viral RNA polymerase. The effect of t-loops depends on the formation of an RNA duplex near the template entry and exit channels of the RNA polymerase, and their effect is enhanced by mutation of the template exit path from the RNA polymerase active site. Overall, these findings provide a mechanism that links aberrant viral replication to the activation of the innate immune response.
]]></description>
<dc:creator>French, H.</dc:creator>
<dc:creator>Pitre, E.</dc:creator>
<dc:creator>Oade, M. S.</dc:creator>
<dc:creator>Elshina, E.</dc:creator>
<dc:creator>Bisht, K.</dc:creator>
<dc:creator>Jahun, A.</dc:creator>
<dc:creator>King, A.</dc:creator>
<dc:creator>Bauer, D. L. V.</dc:creator>
<dc:creator>te Velthuis, A. J. W.</dc:creator>
<dc:date>2022-01-26</dc:date>
<dc:identifier>doi:10.1101/2022.01.25.476955</dc:identifier>
<dc:title><![CDATA[Transient RNA structures cause aberrant influenza virus replication and innate immune activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.01.26.477195v1?rss=1">
<title>
<![CDATA[
Yersinia pestis genomes reveal plague in Britain 4,000 years ago 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.01.26.477195v1?rss=1"
</link>
<description><![CDATA[
Extinct lineages of Yersinia pestis, the causative agent of the plague, have been identified in several individuals from Central Europe and Asia between 5,000 and 3,500 years before present (BP). One of these, the  LNBA lineage (Late Neolithic and Bronze Age), has been suggested to have spread into Central Europe with human groups expanding from the Eurasian steppes. Here, we show that LNBA plague was spread to Europes northwestern periphery by sequencing Yersinia pestis genomes from two individuals dating to ~4,000 cal BP from an unusual mass burial context in Somerset, England, UK. This represents the earliest evidence of plague in Britain documented to date. These British Yersinia pestis genomes belong to a sublineage previously observed in two Bronze Age individuals from Central Europe that had lost the putative virulence factor yapC. This sublineage is later found in Central Asia ~3,600 BP. While the severity of disease is currently unclear, the wide geographic distribution within a few centuries suggests substantial transmissibility.
]]></description>
<dc:creator>Swali, P.</dc:creator>
<dc:creator>Schutling, R.</dc:creator>
<dc:creator>Gilardet, A.</dc:creator>
<dc:creator>Kelly, M.</dc:creator>
<dc:creator>Anastasiadou, K.</dc:creator>
<dc:creator>Glocke, I.</dc:creator>
<dc:creator>Audsley, T.</dc:creator>
<dc:creator>Loe, L.</dc:creator>
<dc:creator>Fernandez-Crespo, T.</dc:creator>
<dc:creator>Ordono, J.</dc:creator>
<dc:creator>Walker, D.</dc:creator>
<dc:creator>Davy, T.</dc:creator>
<dc:creator>Silva, M.</dc:creator>
<dc:creator>Hajdinjak, M.</dc:creator>
<dc:creator>Bergstrom, A.</dc:creator>
<dc:creator>Booth, T.</dc:creator>
<dc:creator>Skoglund, P.</dc:creator>
<dc:date>2022-01-26</dc:date>
<dc:identifier>doi:10.1101/2022.01.26.477195</dc:identifier>
<dc:title><![CDATA[Yersinia pestis genomes reveal plague in Britain 4,000 years ago]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.01.26.477893v1?rss=1">
<title>
<![CDATA[
Multivalent interactions essential for lentiviral integrase function 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.01.26.477893v1?rss=1"
</link>
<description><![CDATA[
A multimer of retroviral integrase (IN) synapses viral DNA ends within a stable intasome nucleoprotein complex for integration into a host cell genome. Reconstitution of the intasome from the maedi-visna virus (MVV), an ovine lentivirus, revealed a large assembly containing sixteen IN subunits (1). Herein, we report cryo-EM structures of the lentiviral intasome prior to engagement of target DNA and following strand transfer, refined at 3.4 and 3.5 [A] resolution, respectively. The structures elucidate details of the protein-protein and protein-DNA interfaces involved in lentiviral intasome formation. We show that the homomeric interfaces involved in IN hexadecamer formation and the -helical configuration of the linker connecting the C-terminal and catalytic core domains are critical for MVV IN strand transfer activity in vitro and for virus infectivity. Single-molecule microscopy in conjunction with photobleaching revealed that the MVV intasome can bind a variable number, up to sixteen molecules, of the lentivirus-specific host factor LEDGF/p75. Concordantly, ablation of endogenous LEDGF/p75 resulted in gross redistribution of MVV integration sites in human and ovine cells. Our data confirm the importance of the expanded architecture observed in cryo-EM studies of lentiviral intasomes and suggest that this organization underlies multivalent interactions with chromatin for integration targeting to active genes.
]]></description>
<dc:creator>Ballandras-Colas, A.</dc:creator>
<dc:creator>Chivukula, V.</dc:creator>
<dc:creator>Gruszka, D. T.</dc:creator>
<dc:creator>Shan, Z.</dc:creator>
<dc:creator>Singh, P. K.</dc:creator>
<dc:creator>Pye, V. E.</dc:creator>
<dc:creator>McLean, R. K.</dc:creator>
<dc:creator>Bedwell, G. J.</dc:creator>
<dc:creator>Li, W.</dc:creator>
<dc:creator>Nans, A.</dc:creator>
<dc:creator>Cook, N. J.</dc:creator>
<dc:creator>Fadel, H. J.</dc:creator>
<dc:creator>Poeschla, E. M.</dc:creator>
<dc:creator>Griffiths, D. J.</dc:creator>
<dc:creator>Vargas, J.</dc:creator>
<dc:creator>Taylor, I. A.</dc:creator>
<dc:creator>Lyumkis, D.</dc:creator>
<dc:creator>Yardimci, H.</dc:creator>
<dc:creator>Engelman, A. N.</dc:creator>
<dc:creator>Cherepanov, P.</dc:creator>
<dc:date>2022-01-27</dc:date>
<dc:identifier>doi:10.1101/2022.01.26.477893</dc:identifier>
<dc:title><![CDATA[Multivalent interactions essential for lentiviral integrase function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.02.08.479664v1?rss=1">
<title>
<![CDATA[
SARS-CoV-2 ORF6 disrupts innate immune signalling by inhibiting cellular mRNA export 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.02.08.479664v1?rss=1"
</link>
<description><![CDATA[
SARS-CoV-2 is a betacoronavirus and the etiological agent of COVID-19, a devastating infectious disease. Due to its far-reaching effect on human health, there is an urgent and growing need to understand the viral molecular biology of SARS-CoV-2 and its interaction with the host cell. SARS-CoV-2 encodes 9 predicted accessory proteins, which are presumed to be dispensable for in vitro replication, most likely having a role in modulating the host cell environment to aid viral replication. Here we show that the ORF6 accessory protein interacts with cellular Rae1 to inhibit cellular protein production by blocking mRNA export. We utilised cell fractionation coupled with mRNAseq to explore which cellular mRNA species are affected by ORF6 expression and show that ORF6 can inhibit the export of many mRNA including those encoding antiviral factors such as IRF1 and RIG-I. We also show that export of these mRNA is blocked in the context of SARS-CoV-2 infection. Together, our studies identify a novel mechanism by which SARS-CoV-2 can manipulate the host cell environment to supress antiviral responses, providing further understanding to the replication strategies of a virus that has caused an unprecedented global health crisis.

Author SummarySARS-CoV-2 is the virus responsible for the current COVID-19 pandemic. Coronaviruses, like SARS-CoV-2, replicate their genome in the cytoplasm of the host cell by hijacking the cellular machinery. In addition to structural proteins and viral enzymes, SARS-CoV-2 encodes 9 accessory proteins. Although these are not required for in vitro replication, they are thought to modulate the host cell environment to favour viral replication. In this work, we show that the ORF6 accessory protein can supress cellular protein production by blocking mRNA nuclear export through interacting with the cellular protein Rae1, a known mRNA export factor. We also investigated which cellular mRNAs were retained in the nucleus when ORF6 was overexpressed. Interestingly, we found that ORF6 inhibited the export of many different mRNAs, including those encoding antiviral factors, like IRF1 and RIG-I, even in the absence of stimulation by interferon. Importantly, we found that the export of these mRNAs was similarly affected in the context of SARS-CoV-2 infection. Therefore, we believe we have identified a novel mechanism that SARS-CoV-2 uses to suppress antiviral responses in order to make the cell more permissive to infection.
]]></description>
<dc:creator>Hall, R.</dc:creator>
<dc:creator>Guedan, A.</dc:creator>
<dc:creator>Yap, M. W.</dc:creator>
<dc:creator>Young, G. R.</dc:creator>
<dc:creator>Harvey, R.</dc:creator>
<dc:creator>Stoye, J. P.</dc:creator>
<dc:creator>Bishop, K. N.</dc:creator>
<dc:date>2022-02-09</dc:date>
<dc:identifier>doi:10.1101/2022.02.08.479664</dc:identifier>
<dc:title><![CDATA[SARS-CoV-2 ORF6 disrupts innate immune signalling by inhibiting cellular mRNA export]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.02.10.477501v1?rss=1">
<title>
<![CDATA[
Lem2 is essential for cardiac development by maintaining nuclear integrity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.02.10.477501v1?rss=1"
</link>
<description><![CDATA[
Nuclear envelope integrity is essential for compartmentalisation of nucleus and cytoplasm. Importantly, mutations in nuclear envelope-encoding genes are the second-highest cause of familial dilated cardiomyopathy. One such nuclear envelope protein that causes cardiomyopathy in humans and affects mouse heart development is Lem2. However, its role in mechanically active tissue such as heart remains poorly understood.

We generated mice in which Lem2 was specifically ablated in cardiomyocytes and carried out detailed physiological, tissue and cellular analyses. Importantly, our data showed that Lem2 was essential for cardiac development, and hearts from Lem2 cKO mice were morphologically and transcriptionally underdeveloped. Lem2 cKO hearts displayed high levels of DNA damage, nuclear rupture, and apoptosis. Crucially, we found that these defects were driven by muscle contraction as they were ameliorated by inhibiting myosin contraction and conversely were exacerbated upon myosin activation.

Our data suggest that Lem2 is critical for integrity at the nascent nuclear envelope in fetal hearts, and protects the nucleus from the mechanical forces of muscle contraction. Taken together, these data provide novel insight into mechanisms underlying striated muscle diseases caused by altered nuclear envelope integrity.
]]></description>
<dc:creator>Ross, J.</dc:creator>
<dc:creator>Arcos-Villacis, N.</dc:creator>
<dc:creator>Battey, E.</dc:creator>
<dc:creator>Boogerd, C.</dc:creator>
<dc:creator>Marhuenda, E.</dc:creator>
<dc:creator>Hodzic, D.</dc:creator>
<dc:creator>Prin, F.</dc:creator>
<dc:creator>Mohun, T.</dc:creator>
<dc:creator>Catibog, N.</dc:creator>
<dc:creator>Tapia, O.</dc:creator>
<dc:creator>Gerace, L.</dc:creator>
<dc:creator>Iskratsch, T.</dc:creator>
<dc:creator>Shah, A.</dc:creator>
<dc:creator>Stroud, M.</dc:creator>
<dc:date>2022-02-10</dc:date>
<dc:identifier>doi:10.1101/2022.02.10.477501</dc:identifier>
<dc:title><![CDATA[Lem2 is essential for cardiac development by maintaining nuclear integrity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.02.13.480269v1?rss=1">
<title>
<![CDATA[
Spindle reorientation in response to mechanical stress is an emergent property of the spindle positioning mechanisms 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.02.13.480269v1?rss=1"
</link>
<description><![CDATA[
Proper orientation of the mitotic spindle plays a crucial role in embryos, during tissue development, and in adults, where it functions to dissipate mechanical stress to maintain tissue integrity and homeostasis. While mitotic spindles have been shown to reorient in response to external mechanical stresses, the subcellular cues that mediate spindle reorientation remain unclear. Here, we have used a combination of optogenetics and computational modelling to better understand how mitotic spindles respond to inhomogeneous tension within the actomyosin cortex. Strikingly, we find that the optogenetic activation of RhoA only influences spindle orientation when it is induced at both poles of the cell. Under these conditions, the sudden local increase in cortical tension induced by RhoA activation reduces pulling forces exerted by cortical regulators on astral microtubules. This leads to a perturbation of the torque balance exerted on the spindle, which causes it to rotate. Thus, spindle rotation in response to mechanical stress is an emergent phenomenon arising from the interaction between the spindle positioning machinery and the cell cortex.
]]></description>
<dc:creator>Kelkar, M.</dc:creator>
<dc:creator>Bohec, P.</dc:creator>
<dc:creator>Smith, M.</dc:creator>
<dc:creator>Sreenivasan, V.</dc:creator>
<dc:creator>Lisica, A.</dc:creator>
<dc:creator>Valon, L.</dc:creator>
<dc:creator>Ferber, E.</dc:creator>
<dc:creator>Baum, B.</dc:creator>
<dc:creator>Salbreux, G.</dc:creator>
<dc:creator>Charras, G.</dc:creator>
<dc:date>2022-02-14</dc:date>
<dc:identifier>doi:10.1101/2022.02.13.480269</dc:identifier>
<dc:title><![CDATA[Spindle reorientation in response to mechanical stress is an emergent property of the spindle positioning mechanisms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.02.15.480567v1?rss=1">
<title>
<![CDATA[
A gradient border model for cell fate decisions at the neural plate border 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.02.15.480567v1?rss=1"
</link>
<description><![CDATA[
The vertebrate  neural plate border is a transient territory located at the edge of the neural plate containing precursors for all ectodermal derivatives: the neural plate; neural crest; placodes; and epidermis. Elegant functional experiments in a range of vertebrate models have provided an in-depth understanding of gene regulatory interactions within the ectoderm. However, these experiments conducted at tissue level raise seemingly contradictory models for fate allocation of individual cells. Here, we carry out single cell RNA sequencing of chick ectoderm from primitive streak to neurulation stage, to explore cell state diversity and heterogeneity. We characterise the dynamics of gene modules containing key factors known to regulate ectodermal cell fates, allowing us to model the order in which these fates are specified. Furthermore, we find that genes previously classified as neural plate border specifiers typically exhibit dynamic expression patterns and are biased towards either placodal or neural crest fates, revealing that the neural plate border should be seen as an anatomical region of the ectoderm and not a discrete transcriptional state. Through co-expression of placodal and neural crest markers, we identify a population of border located unstable progenitors (BLUPs) which gradually reduces in size as fate segregation occurs. Considering our findings, we propose a  gradient border model for cell fate choice at the neural plate border, with the probability of cell fate allocation closely tied to the spatiotemporal positioning of cells.
]]></description>
<dc:creator>Thiery, A.</dc:creator>
<dc:creator>Buzzi, A. L.</dc:creator>
<dc:creator>Hamrud, E.</dc:creator>
<dc:creator>Cheshire, C.</dc:creator>
<dc:creator>Luscombe, N.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Streit, A.</dc:creator>
<dc:date>2022-02-16</dc:date>
<dc:identifier>doi:10.1101/2022.02.15.480567</dc:identifier>
<dc:title><![CDATA[A gradient border model for cell fate decisions at the neural plate border]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.02.17.480906v1?rss=1">
<title>
<![CDATA[
Clonal behaviour of myogenic precursor cells throughout the vertebrate lifespan. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.02.17.480906v1?rss=1"
</link>
<description><![CDATA[
To address questions of stem cell diversity during skeletal myogenesis, a Brainbow-like genetic cell lineage tracing method, dubbed Musclebow, was derived by enhancer trapping in zebrafish. It is shown that at least 15 muscle precursor cells (mpcs) seed each somite, where they proliferate but contribute little to muscle growth prior to hatching. Thereafter, dermomyotome-derived mpc clones rapidly expand while some progeny undergo terminal differentiation, leading to stochastic clonal drift. No evidence of cell lineage-based clonal fate diversity was obtained. Neither fibre nor mpc death was observed in uninjured animals. Individual marked muscle fibres persist across much of the lifespan indicating low rates of nuclear turnover. In adulthood, early-marked mpc clones label stable blocks of tissue comprising a significant fraction of either epaxial or hypaxial somite. Fusion of cells from separate early-marked clones occurs in regions of clone overlap. Wounds are regenerated from many/most local mpcs; no evidence for specialised stem mpcs was obtained. In conclusion, our data indicate that most mpcs in muscle tissue contribute to local growth and repair and suggest that cellular turnover is low in the absence of trauma.

Summary StatementMusclebow clonal cell lineage analysis is introduced to reveal the cellular dynamics of skeletal muscle formation, repair and maintenance throughout the life of zebrafish.
]]></description>
<dc:creator>Hughes, S. M.</dc:creator>
<dc:creator>Escaleira, R. C.</dc:creator>
<dc:creator>Wanders, K.</dc:creator>
<dc:creator>Koth, J.</dc:creator>
<dc:creator>Wilkinson, D. G.</dc:creator>
<dc:creator>Xu, Q.</dc:creator>
<dc:date>2022-02-18</dc:date>
<dc:identifier>doi:10.1101/2022.02.17.480906</dc:identifier>
<dc:title><![CDATA[Clonal behaviour of myogenic precursor cells throughout the vertebrate lifespan.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.02.18.481045v1?rss=1">
<title>
<![CDATA[
Sample preparation and warping accuracy for correlative multimodal imaging in the mouse olfactory bulb using 2-photon, synchrotron X-ray and volume electron microscopy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.02.18.481045v1?rss=1"
</link>
<description><![CDATA[
Integrating physiology with structural insights of the same neuronal circuit provides a unique approach to understanding how the mammalian brain computes information. However, combining the techniques that provide both streams of data represents an experimental challenge. When studying glomerular column circuits in the mouse olfactory bulb, this approach involves e.g. recording the neuronal activity with in vivo 2-photon (2P) calcium imaging, retrieving the circuit structure with synchrotron X-ray computed tomography with propagation-based phase contrast (SXRT) and/or serial block-face electron microscopy (SBEM) and correlating these datasets. Sample preparation and dataset correlation are two key bottlenecks in this correlative workflow. Here, we first quantify the occurrence of different artefacts when staining tissue slices with heavy metals to generate X-ray or electron contrast. We report improvements in the staining procedure, ultimately achieving perfect staining in [~]67% of the 0.6 mm thick olfactory bulb slices that were previously imaged in vivo with 2P. Secondly, we characterise the accuracy of the spatial correlation between functional and structural datasets. We demonstrate that direct, single-cell precise correlation between in vivo 2P and SXRT tissue volumes is possible and as reliable as correlating between 2P and SBEM. Altogether, these results pave the way for experiments that require retrieving physiology, circuit structure and synaptic signatures in targeted regions. These correlative function-structure studies will bring a more complete understanding of mammalian olfactory processing across length scales and time.
]]></description>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Ackels, T.</dc:creator>
<dc:creator>Pacureanu, A.</dc:creator>
<dc:creator>Zdora, M.-C.</dc:creator>
<dc:creator>Bonnin, A.</dc:creator>
<dc:creator>Schaefer, A. T.</dc:creator>
<dc:creator>Bosch, C.</dc:creator>
<dc:date>2022-02-20</dc:date>
<dc:identifier>doi:10.1101/2022.02.18.481045</dc:identifier>
<dc:title><![CDATA[Sample preparation and warping accuracy for correlative multimodal imaging in the mouse olfactory bulb using 2-photon, synchrotron X-ray and volume electron microscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.02.22.481424v1?rss=1">
<title>
<![CDATA[
Combining SLA 3D printing and soft lithography for fast, versatile, and accessible high-resolution fabrication of customised multiscale cell culture devices with complex designs 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.02.22.481424v1?rss=1"
</link>
<description><![CDATA[
Cell culture devices, such as microwells and microfluidic chips, are designed to increase the complexity of cell-based models whilst retaining control over culture conditions and have become indispensable platforms for biological systems modelling. From microtopography, microwells, plating devices and microfluidic systems to larger constructs for specific applications such as live imaging chamber slides, a wide variety of culture devices with different geometries have become indispensable in biology laboratories. However, while their application in biological projects is increasing exponentially, due to a combination of the techniques and tools required for their manufacture, and the physical science background sometimes needed, the design and fabrication of such devices directly by biological labs remains a relatively high investment in terms of costs, use of facilities, needed collaborations and time. Whilst commercially available systems are available, these are also often costly, and importantly lack the potential for customisation by each single lab. This combination of factors still limits widespread application of microfabricated custom devices in most biological wet labs.

Capitalising on recent important advancements in the fields of bioengineering and microfabrication, and taking advantage of low-cost, high-resolution desktop resin 3D printers combined with PDMS soft lithography, we have developed an optimised low-cost and highly reproducible microfabrication pipeline, capable of generating a wide variety of customisable devices for cell culture and tissue engineering in an easy, fast reproducible way for a fraction of the cost of conventional microfabrication or commercial alternatives. This protocol is designed specifically to be a resource for biological labs with little to none prior exposure to these fields technique and enables the manufacture of complex devices across the {micro}m to cm scale.

We provide a ready-to-go pipeline for the efficient treatment of resin-based 3D printed constructs for PDMS curing, using a combination of curing steps, washes and surface treatments. Together with the extensive characterisation of the fabrication pipeline, we show the utilization of this system to a variety of applications and use cases relevant to biological experiments, ranging from micro topographies for cell alignments to complex multi-part hydrogel culturing systems.

This methodology can be easily adopted by any wet lab, irrespective of prior expertise or resource availability and will enable the wide adoption of tailored microfabricated devices across many fields of biology.
]]></description>
<dc:creator>Hagemann, C.</dc:creator>
<dc:creator>Bailey, M. C. D.</dc:creator>
<dc:creator>Khokhar, N.</dc:creator>
<dc:creator>Lionello, V. M.</dc:creator>
<dc:creator>Suklai, P.</dc:creator>
<dc:creator>Moreno-Gonzalez, C.</dc:creator>
<dc:creator>O'Toole, K.</dc:creator>
<dc:creator>Konstantinou, G.</dc:creator>
<dc:creator>Giagnorio, E.</dc:creator>
<dc:creator>Bergholt, M.</dc:creator>
<dc:creator>Imbert, A.</dc:creator>
<dc:creator>Tedesco, F. S.</dc:creator>
<dc:creator>Serio, A.</dc:creator>
<dc:date>2022-02-23</dc:date>
<dc:identifier>doi:10.1101/2022.02.22.481424</dc:identifier>
<dc:title><![CDATA[Combining SLA 3D printing and soft lithography for fast, versatile, and accessible high-resolution fabrication of customised multiscale cell culture devices with complex designs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.07.483228v1?rss=1">
<title>
<![CDATA[
Cell-cell metabolite exchange creates a pro-survival metabolic environment that extends lifespan 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.07.483228v1?rss=1"
</link>
<description><![CDATA[
Metabolism is fundamentally intertwined with the ageing process. We here report that a key determinant of cellular lifespan is not only nutrient supply and intracellular metabolism, but also metabolite exchange interactions that occur between cells. Studying chronological ageing in yeast, we observed that metabolites exported by young, exponentially growing, cells are re- imported during the stationary phase when cells age chronologically, indicating the existence of cross-generational metabolic interactions. We then used self-establishing metabolically cooperating communities (SeMeCos) to boost cell-cell metabolic interactions and observed a significant lifespan extension. A search for the underlying mechanisms, coupling SeMeCos, metabolic profiling, proteomics and genome-scale metabolic modelling, attributed a specific role to methionine consumer cells. These cells were enriched over time, adopted glycolytic metabolism and increased export of protective metabolites. Glycerol, in particular, accumulated in the communal metabolic environment and extended the lifespan of all cells in the community in a paracrine fashion. Our results hence establish metabolite exchange interactions as a determinant of the ageing process and show that metabolically cooperating cells shape their metabolic environment to achieve lifespan extension.
]]></description>
<dc:creator>Correia-Melo, C.</dc:creator>
<dc:creator>Kamrad, S.</dc:creator>
<dc:creator>Messner, C. B.</dc:creator>
<dc:creator>Tengolics, R.</dc:creator>
<dc:creator>Herrera-Dominguez, L.</dc:creator>
<dc:creator>Townsend, S.</dc:creator>
<dc:creator>Alam, M. T.</dc:creator>
<dc:creator>Freiwald, A.</dc:creator>
<dc:creator>Campbell, K.</dc:creator>
<dc:creator>Aulakh, S.</dc:creator>
<dc:creator>Szyrwiel, L.</dc:creator>
<dc:creator>Yu, J. S. L.</dc:creator>
<dc:creator>Zelezniak, A.</dc:creator>
<dc:creator>Demichev, V.</dc:creator>
<dc:creator>Muelleder, M.</dc:creator>
<dc:creator>Papp, B.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:date>2022-03-07</dc:date>
<dc:identifier>doi:10.1101/2022.03.07.483228</dc:identifier>
<dc:title><![CDATA[Cell-cell metabolite exchange creates a pro-survival metabolic environment that extends lifespan]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.09.483611v1?rss=1">
<title>
<![CDATA[
Diet suppresses tumour initiation by maintaining quiescence of mutation-bearing neural stem cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.09.483611v1?rss=1"
</link>
<description><![CDATA[
Glioblastoma is thought to originate from neural stem cells (NSCs) of the subventricular zone that acquire genetic alterations. In the adult brain, NSCs are largely quiescent, suggesting that deregulation of quiescence maintenance may be a pre-requisite for tumour initiation. Although inactivation of the tumour suppressor p53 is a frequent event in gliomagenesis, whether, or how, it affects quiescent NSCs (qNSCs) remains unclear. Here we show that p53 maintains quiescence by inducing fatty acid oxidation (FAO) and that acute p53 deletion in qNSCs results in their premature activation to a proliferative state. Mechanistically, this occurs through direct transcriptional induction of PPARGC1a, which in turn activates PPAR to upregulate FAO genes. Strikingly, dietary supplementation with fish oil containing omega-3 fatty acids, natural PPAR ligands, fully restores quiescence of p53-deficient NSCs and delays tumour initiation in a glioblastoma mouse model. Thus, diet can silence glioblastoma driver mutations, with important implications for cancer prevention.
]]></description>
<dc:creator>Amodeo, V.</dc:creator>
<dc:creator>Davies, T.</dc:creator>
<dc:creator>Martinez-Segura, A.</dc:creator>
<dc:creator>Clements, M.</dc:creator>
<dc:creator>Simpson Ragdale, H.</dc:creator>
<dc:creator>Bailey, A.</dc:creator>
<dc:creator>Silva dos Santos, M.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Mokochinski, J.</dc:creator>
<dc:creator>Kramer, H.</dc:creator>
<dc:creator>Garcia-Diaz, C.</dc:creator>
<dc:creator>Gould, A. P.</dc:creator>
<dc:creator>Marguerat, S.</dc:creator>
<dc:creator>Parrinello, S.</dc:creator>
<dc:date>2022-03-11</dc:date>
<dc:identifier>doi:10.1101/2022.03.09.483611</dc:identifier>
<dc:title><![CDATA[Diet suppresses tumour initiation by maintaining quiescence of mutation-bearing neural stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.11.483935v1?rss=1">
<title>
<![CDATA[
Hypoxia shapes the immune landscape in lung injury promoting inflammation persistence 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.11.483935v1?rss=1"
</link>
<description><![CDATA[
Acute Respiratory Distress Syndrome (ARDS), an often-fatal complication of pulmonary or systemic inflammation, has no cure. Hypoxemia is a defining feature, yet its impact on inflammation is often neglected. Patients with ARDS are monocytopenic early in the onset of the disease. Endotoxin or Streptococcus pneumoniae acute lung injury (ALI) in the context of hypoxia replicates this finding, through hypoxia-driven suppression of type I interferon signalling. This results in failed lung monocyte-derived interstitial macrophages (IM) niche expansion and unchecked neutrophilic inflammation. Administration of colony stimulating factor 1 (CSF1) rescues the monocytopenia, alters the circulating classical monocyte phenotype in hypoxic endotoxin-driven ALI and enables lung IM population expansion, thus limiting lung injury in endotoxin- and virally-induced hypoxic ALI. Hypoxia directly alters immune dynamics to the detriment of the host and manipulation of this aberrant response offers new therapeutic strategies for ARDS.
]]></description>
<dc:creator>Mirchandani, A. S.</dc:creator>
<dc:creator>Jenkins, S. J.</dc:creator>
<dc:creator>Bain, C. C.</dc:creator>
<dc:creator>Lawson, H.</dc:creator>
<dc:creator>Coelho, P.</dc:creator>
<dc:creator>Murphy, F.</dc:creator>
<dc:creator>Griffith, D.</dc:creator>
<dc:creator>Zhang, A.</dc:creator>
<dc:creator>Sanchez-Garcia, M. A.</dc:creator>
<dc:creator>Reyes, L.</dc:creator>
<dc:creator>Morrison, T.</dc:creator>
<dc:creator>Arienti, S.</dc:creator>
<dc:creator>Sadiku, P.</dc:creator>
<dc:creator>Watts, E.</dc:creator>
<dc:creator>Dickinson, R.</dc:creator>
<dc:creator>Clark, S.</dc:creator>
<dc:creator>Ly, T.</dc:creator>
<dc:creator>Lewis, D.</dc:creator>
<dc:creator>Kelly, V.</dc:creator>
<dc:creator>Spanos, C.</dc:creator>
<dc:creator>Musgrave, K.</dc:creator>
<dc:creator>Delaney, L.</dc:creator>
<dc:creator>Harper, I.</dc:creator>
<dc:creator>Scott, J.</dc:creator>
<dc:creator>Parkinson, N.</dc:creator>
<dc:creator>Rostron, A.</dc:creator>
<dc:creator>Baillie, J. K.</dc:creator>
<dc:creator>Clohisey, S. M. R.</dc:creator>
<dc:creator>Pridans, C.</dc:creator>
<dc:creator>Campana, L.</dc:creator>
<dc:creator>Starkey, P.</dc:creator>
<dc:creator>Simpson, A. J.</dc:creator>
<dc:creator>Dockrell, D.</dc:creator>
<dc:creator>Schwarze, J.</dc:creator>
<dc:creator>Hirani, N.</dc:creator>
<dc:creator>Ratcliffe, P.</dc:creator>
<dc:creator>Pugh, C.</dc:creator>
<dc:creator>Kranc, K.</dc:creator>
<dc:creator>Forbes, S.</dc:creator>
<dc:creator>Whyte, M.</dc:creator>
<dc:creator>Walmsley, S.</dc:creator>
<dc:date>2022-03-12</dc:date>
<dc:identifier>doi:10.1101/2022.03.11.483935</dc:identifier>
<dc:title><![CDATA[Hypoxia shapes the immune landscape in lung injury promoting inflammation persistence]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.10.483782v1?rss=1">
<title>
<![CDATA[
Genetic mapping of APP and amyloid-β biology modulation by trisomy 21 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.10.483782v1?rss=1"
</link>
<description><![CDATA[
Individuals who have Down syndrome frequently develop early onset Alzheimers disease, a neurodegenerative condition caused by the build-up of aggregated amyloid-{beta} and tau proteins in the brain. Amyloid-{beta} is produced by APP, a gene located on chromosome 21. People who have Down syndrome have three copies of chromosome 21 and thus also an additional copy of APP; this genetic change drives the early development of Alzheimers disease in these individuals. Here we use a combination of next-generation mouse models of Down syndrome (Tc1, Dp3Tyb, Dp(10)2Yey and Dp(17)3Yey) and a knockin mouse model of amyloid-{beta} accumulation (AppNL-F) to determine how chromosome 21 genes other than APP modulate APP/amyloid-{beta} in the brain when in three copies. We demonstrate that three copies of other chromosome 21 genes are sufficient to partially ameliorate amyloid-{beta} accumulation in the brain. We go on to identify a subregion of chromosome 21 that contains the gene/genes causing this decrease in amyloid-{beta} accumulation and investigate the role of two lead candidate genes Dyrk1a and Bace2. Thus an additional copy of chromosome 21 genes, other than APP, can modulate APP/amyloid-{beta} in the brain under physiological conditions. This work provides critical mechanistic insight into the development of disease and an explanation for the typically later age of onset of dementia in people who have AD-DS compared to those who have familial AD caused by triplication of APP.
]]></description>
<dc:creator>Mumford, P.</dc:creator>
<dc:creator>Tosh, J.</dc:creator>
<dc:creator>Anderle, S.</dc:creator>
<dc:creator>Wikberg, E. G.</dc:creator>
<dc:creator>Lau, G.</dc:creator>
<dc:creator>Noy, S.</dc:creator>
<dc:creator>Cleverley, K.</dc:creator>
<dc:creator>Saito, T.</dc:creator>
<dc:creator>Saido, T. C.</dc:creator>
<dc:creator>Yu, E. Y.</dc:creator>
<dc:creator>Brinkmalm, G.</dc:creator>
<dc:creator>Portelius, E.</dc:creator>
<dc:creator>Blennow, K.</dc:creator>
<dc:creator>Zetterberg, H.</dc:creator>
<dc:creator>Tybulewicz, V.</dc:creator>
<dc:creator>Fisher, E.</dc:creator>
<dc:creator>Wiseman, F. K.</dc:creator>
<dc:date>2022-03-12</dc:date>
<dc:identifier>doi:10.1101/2022.03.10.483782</dc:identifier>
<dc:title><![CDATA[Genetic mapping of APP and amyloid-β biology modulation by trisomy 21]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.10.482611v1?rss=1">
<title>
<![CDATA[
Placental inflammation leads to abnormal embryonic heart development 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.10.482611v1?rss=1"
</link>
<description><![CDATA[
Placental and embryonic heart development occurs in parallel, and these organs have been proposed to exert reciprocal regulation during gestation. Poor placentation has been associated with congenital heart disease (CHD), an important cause of infant mortality. However, the mechanisms by which altered placental development can lead to CHD remain unresolved. In the current study we show that neutrophil-driven placental inflammation leads to inadequate placental development and loss of barrier function. Consequently, placental inflammatory monocytes of maternal origin become capable to migrate to the embryonic heart and alter the normal composition of resident cardiac macrophages and cardiac tissue structure. This cardiac impairment continues into postnatal life, hindering normal tissue architecture and function. Finally, we demonstrate that tempering placental inflammation can rescue this fetal cardiac defect and is sufficient to promote normal cardiac function in postnatal life. Taken together, our observations provide a mechanistic paradigm whereby neutrophil-driven inflammation in pregnancy can preclude normal embryonic heart development as a direct consequence of poor placental development.
]]></description>
<dc:creator>Ward, E. J.</dc:creator>
<dc:creator>Bert, S.</dc:creator>
<dc:creator>Fanti, S.</dc:creator>
<dc:creator>Dufton, N. P.</dc:creator>
<dc:creator>Malone, K. M.</dc:creator>
<dc:creator>Maughan, R. T.</dc:creator>
<dc:creator>Prin, F.</dc:creator>
<dc:creator>Volpato, L.-K.</dc:creator>
<dc:creator>Piovesan, A. P.</dc:creator>
<dc:creator>Perretti, M.</dc:creator>
<dc:creator>Marelli-Berg, F. M.</dc:creator>
<dc:creator>Nadkarni, S.</dc:creator>
<dc:date>2022-03-12</dc:date>
<dc:identifier>doi:10.1101/2022.03.10.482611</dc:identifier>
<dc:title><![CDATA[Placental inflammation leads to abnormal embryonic heart development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.10.482156v1?rss=1">
<title>
<![CDATA[
Deep learning in human neurons predicts mechanistic subtypes of Parkinson's 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.10.482156v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is a common, devastating, and incurable neurodegenerative disorder. Several molecular mechanisms have been proposed to drive PD, with genetic and pathological evidence pointing towards aberrant protein homeostasis and mitochondrial dysfunction. PD is clinically highly heterogeneous, it is likely that different mechanisms underlie the pathology in different individuals, each requiring a specific targeted treatment. Recent advances in stem cell technology and fluorescent live-cell imaging have enabled the generation of patient-derived neurons with different mechanistic subtypes of PD. Here, we performed multi-dimensional fluorescent labelling of organelles in iPSC-derived neurons, in healthy control cells, and in four different disease subclasses. We generated a machine learning-based model that can simultaneously predict the presence of disease, and its primary mechanistic subtype. We independently trained a series of classifiers using both quantitative single-cell fluorescence variables and images to build deep neural networks. Quantitative cellular profile-based classifiers achieve an accuracy of 82%, whilst image based deep neural networks predict control, and four distinct disease subtypes with an accuracy of 95%. The classifiers achieve their accuracy across all subtypes primarily utilizing the organellar features of the mitochondria, with additional contribution of the lysosomes, confirming their biological importance in PD. Taken together, we show that machine learning approaches applied to patient-derived cells are able to predict disease subtypes, demonstrating that this approach may be used to guide personalized treatment approaches in the future.
]]></description>
<dc:creator>D'Sa, K.</dc:creator>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>Virdi, G. S.</dc:creator>
<dc:creator>Vecchi, G.</dc:creator>
<dc:creator>Adam, A.</dc:creator>
<dc:creator>Bertolli, O.</dc:creator>
<dc:creator>Fleming, J.</dc:creator>
<dc:creator>Chang, H.</dc:creator>
<dc:creator>Athauda, D.</dc:creator>
<dc:creator>Choi, M. L.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:date>2022-03-12</dc:date>
<dc:identifier>doi:10.1101/2022.03.10.482156</dc:identifier>
<dc:title><![CDATA[Deep learning in human neurons predicts mechanistic subtypes of Parkinson's]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.13.484137v1?rss=1">
<title>
<![CDATA[
Systematic dissection of phosphorylation-dependent YAP1 complex formation elucidates a key role for PTPN14 in Hippo signal integration 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.13.484137v1?rss=1"
</link>
<description><![CDATA[
Cellular signaling relies on the temporal and spatial control of the formation of transient protein complexes by post-translational modifications, most notably by phosphorylation. While several computational methods have been developed to predict the functional relevance of phosphorylation sites, assessing experimentally the interdependency between protein phosphorylation and protein-protein interactions (PPIs) remains a major challenge. Here, we describe an experimental strategy to establish interdependencies between specific phosphorylation events and complex formation. This strategy is based on three main steps: (i) systematically charting the phosphorylation landscape of a target protein; (ii) assigning distinct proteoforms of the target protein to different protein complexes by electrophoretic separation of native complexes (BNPAGE) and protein/phopho correlation profiling; and (iii) genetically deleting known regulators of the target protein to identify which ones are required for given proteoforms and complexes. We applied this strategy to study phosphorylation- dependent modulation of complexes containing the transcriptional co-regulator YAP1. YAP1 is highly phosphorylated and among the most extensively connected proteins in the human interactome. It functions as the main signal integrator and effector protein of the Hippo pathway which controls organ size and tissue homeostasis. Using our workflow, we could identify several distinct YAP1 proteoforms specifically associated with physically distinct complexes and infer how their formation is affected by known Hippo pathway members. Importantly, our findings suggest that the tyrosine phosphatase PTPN14 controls the co-transcriptional activity of YAP1 by regulating its interaction with the LATS1/2 kinases. In summary, we present a powerful strategy to establish interdependencies between specific phosphorylation events and complex formation, thus contributing to the "functionalization" of phosphorylation events and by this means provide new insights into Hippo signaling.
]]></description>
<dc:creator>Uliana, F.</dc:creator>
<dc:creator>Ciuffa, R.</dc:creator>
<dc:creator>Mishra, R.</dc:creator>
<dc:creator>Fossati, A.</dc:creator>
<dc:creator>Frommelt, F.</dc:creator>
<dc:creator>Mehnert, M.</dc:creator>
<dc:creator>Salmorin, E. B.</dc:creator>
<dc:creator>Peter, M.</dc:creator>
<dc:creator>Tapon, N.</dc:creator>
<dc:creator>Aebersold, R.</dc:creator>
<dc:creator>Gstaiger, M.</dc:creator>
<dc:date>2022-03-13</dc:date>
<dc:identifier>doi:10.1101/2022.03.13.484137</dc:identifier>
<dc:title><![CDATA[Systematic dissection of phosphorylation-dependent YAP1 complex formation elucidates a key role for PTPN14 in Hippo signal integration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.14.484209v1?rss=1">
<title>
<![CDATA[
Inorganic sulfur fixation via a new homocysteine synthase allows yeast cells to cooperatively compensate for methionine auxotrophy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.14.484209v1?rss=1"
</link>
<description><![CDATA[
The assimilation, incorporation, and metabolism of sulfur is a fundamental process across all domains of life, yet how cells deal with varying sulfur availability is not well understood. We studied an unresolved conundrum of sulfur fixation in yeast, in which an organosulfur-auxotrophy caused by deletion of homocysteine synthase Met17p is overcome when cells are inoculated at high cell density. We discovered that an uncharacterized gene YLL058Wp, herein named Hydrogen sulfide utilizing-1 (HSU1), acts as a homocysteine synthase and allows the cells to substitute for Met17p by re-assimilating hydrosulfide ions leaked from met17{Delta} cells into O-acetyl-homoserine and forming homocysteine. Our results show that cells can cooperate to achieve sulfur fixation, indicating that the collective properties of microbial communities facilitate their basic metabolic capacity.

SummarySulfur limitation activates a dormant hydrogen sulfide fixation route via a novel homocysteine synthase Hsu1p (YLL058Wp).
]]></description>
<dc:creator>Yu, J. S.</dc:creator>
<dc:creator>Heineike, B. M.</dc:creator>
<dc:creator>Hartl, J.</dc:creator>
<dc:creator>Correia-Melo, C.</dc:creator>
<dc:creator>Aulakh, S. K.</dc:creator>
<dc:creator>Lehmann, A.</dc:creator>
<dc:creator>Lemke, O.</dc:creator>
<dc:creator>Agostini, F.</dc:creator>
<dc:creator>Lee, C. T.</dc:creator>
<dc:creator>Demichev, V.</dc:creator>
<dc:creator>Messner, C. B.</dc:creator>
<dc:creator>Mülleder, M.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:date>2022-03-14</dc:date>
<dc:identifier>doi:10.1101/2022.03.14.484209</dc:identifier>
<dc:title><![CDATA[Inorganic sulfur fixation via a new homocysteine synthase allows yeast cells to cooperatively compensate for methionine auxotrophy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.16.484584v1?rss=1">
<title>
<![CDATA[
T cell migration requires ion and water influx to regulate actin polymerization 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.16.484584v1?rss=1"
</link>
<description><![CDATA[
Migration of T cells is essential for their ability to mount immune responses. Chemokine-induced T cell migration requires WNK1, a kinase that regulates ion influx into the cell. However, it is not known why ion entry is necessary for T cell movement. Here we show that signaling from the chemokine receptor CCR7 leads to activation of WNK1 and its downstream pathway at the leading edge of migrating CD4+ T cells, resulting in ion influx and water entry by osmosis. We propose that WNK1-induced water entry is required to swell the membrane at the leading edge, generating space into which actin filaments can polymerize, thereby facilitating forward movement of the cell. Given the broad expression of WNK1 pathway proteins, our study suggests that ion and water influx are likely to be essential for migration in many cell types, including leukocytes and metastatic tumor cells.

One Sentence SummaryChemokine-induced migration of T cells requires water entry at the leading edge to facilitate actin polymerization.
]]></description>
<dc:creator>de Boer, L.</dc:creator>
<dc:creator>Melgrati, S.</dc:creator>
<dc:creator>Vanes, L.</dc:creator>
<dc:creator>Biggs O'May, J.</dc:creator>
<dc:creator>Hayward, D.</dc:creator>
<dc:creator>Koechl, R.</dc:creator>
<dc:creator>Tybulewicz, V.</dc:creator>
<dc:date>2022-03-18</dc:date>
<dc:identifier>doi:10.1101/2022.03.16.484584</dc:identifier>
<dc:title><![CDATA[T cell migration requires ion and water influx to regulate actin polymerization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.16.484637v1?rss=1">
<title>
<![CDATA[
Water influx is required for CD4+ T cell activation and T cell-dependent antibody responses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.16.484637v1?rss=1"
</link>
<description><![CDATA[
Signaling from the T cell antigen receptor (TCR) on CD4+ T cells plays a critical role in adaptive immune responses by inducing T cell activation, proliferation, and differentiation. We demonstrate that WNK1, a kinase implicated in osmoregulation in the kidney, is required in T cells to support T-dependent antibody responses. WNK1-deficient CD4+ T cells are severely impaired in their ability to proliferate and to generate antigen-specific T follicular helper cells in response to immunization with a T-dependent antigen. We show that WNK1 and its downstream OXSR1 and STK39 kinases are required for TCR signaling in CD4+ T cells and for entry into cell cycle. Additionally, by preventing ATR activation, this pathway is required for T cells to progress from G2 into M phase of the cell cycle. Unexpectedly, we show that this WNK1 pathway regulates water influx, most likely through AQP3, which is required for TCR-induced signaling and cell cycle entry. Thus, TCR signaling via WNK1, OXSR1, STK39 and AQP3 leads to water entry that is essential for CD4+ T cell proliferation and hence T cell-dependent antibody responses. Given the broad expression of WNK1, WNK1-dependent water influx may be a common feature of mitogenic pathways in many cell types, both within the immune system and beyond.

One Sentence SummaryT cell antigen receptor signaling via the WNK1 kinase causes water entry which is essential for CD4+ T cell proliferation.
]]></description>
<dc:creator>Biggs O'May, J.</dc:creator>
<dc:creator>Vanes, L.</dc:creator>
<dc:creator>Hartweger, H.</dc:creator>
<dc:creator>de Boer, L.</dc:creator>
<dc:creator>Hayward, D.</dc:creator>
<dc:creator>Koechl, R.</dc:creator>
<dc:creator>Tybulewicz, V.</dc:creator>
<dc:date>2022-03-18</dc:date>
<dc:identifier>doi:10.1101/2022.03.16.484637</dc:identifier>
<dc:title><![CDATA[Water influx is required for CD4+ T cell activation and T cell-dependent antibody responses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.18.484894v1?rss=1">
<title>
<![CDATA[
Molecular determinants of the Bacillus subtilis chromosome origin basal unwinding system 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.18.484894v1?rss=1"
</link>
<description><![CDATA[
Genome duplication is essential for cell proliferation and DNA synthesis is generally initiated by dedicated replication proteins at specific loci termed origins. During DNA replication initiation in bacteria, the ubiquitous DnaA protein engages both double-strand DNA (dsDNA) and single-stranded DNA (ssDNA) at the chromosome origin (oriC) to promote DNA duplex unwinding. While the molecular basis for DnaA binding to a specific dsDNA element ("DnaA-box") has been established, the mechanism for DnaA binding to a specific ssDNA motif ("DnaA-trio") is unclear. Here we define specific steps of DnaA-trio engagement by Bacillus subtilis DnaA. Single-molecule total internal reflection fluorescence microscopy indicates that DnaA proteins are loaded onto DnaA-trios using DnaA-boxes located on a shared DNA polymer. Chemical modification of either the phosphodiester backbone or the nucleobases revealed that three DnaA-trio repeats proximal to DnaA-boxes are necessary and sufficient to promote DnaA-dependent strand separation, and that the amino group from the central nucleobase of the DnaA-trio is critical for this reaction. Finally, based on electrophoretic mobility shift assays, we propose that during replication initiation DnaA progresses from DnaA-boxes to nucleobase recognition at DnaA-trios before engaging the phosphodiester backbone and destabilizing the DNA duplex. These results provide new molecular insight into DnaA-dependent Bacterial Unwinding System (BUS) activity at a bacterial chromosome origin.
]]></description>
<dc:creator>Pelliciari, S.</dc:creator>
<dc:creator>Burnham, D. R.</dc:creator>
<dc:creator>Merces, G.</dc:creator>
<dc:creator>Yardimci, H.</dc:creator>
<dc:creator>Murray, H.</dc:creator>
<dc:date>2022-03-19</dc:date>
<dc:identifier>doi:10.1101/2022.03.18.484894</dc:identifier>
<dc:title><![CDATA[Molecular determinants of the Bacillus subtilis chromosome origin basal unwinding system]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.21.485090v1?rss=1">
<title>
<![CDATA[
Toxoplasma gondii ROP1 subverts murine and human innate immune restriction 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.21.485090v1?rss=1"
</link>
<description><![CDATA[
Toxoplasma gondii is an intracellular parasite that can infect many different host species and is a cause of significant human morbidity worldwide. T. gondii secretes a diverse array of effector proteins into the host cell which are critical for infection; however, the vast majority of these secreted proteins are uncharacterised. Here, we carried out a pooled CRISPR knockout screen in the T. gondii Prugniaud strain in vivo to identify secreted proteins that contribute to parasite immune evasion in the host. We identify 22 putative virulence factors and demonstrate that ROP1, the first-identified rhoptry protein of T. gondii, has a previously unrecognised role in parasite resistance to interferon gamma-mediated innate immune restriction. This function is conserved in the highly virulent RH strain of T. gondii and contributes to parasite growth in both murine and human macrophages. While ROP1 affects the morphology of rhoptries, from where the protein is secreted, it does not affect rhoptry secretion. ROP1 interacts with the host cell protein C1QBP, which appears to facilitate parasite immune evasion. In summary, we identify 22 secreted proteins which contribute to parasite growth in vivo and show that ROP1 is an important and previously overlooked effector in counteracting both murine and human innate immunity.
]]></description>
<dc:creator>Butterworth, S.</dc:creator>
<dc:creator>Torelli, F.</dc:creator>
<dc:creator>Lockyer, E. J.</dc:creator>
<dc:creator>Wagener, J.</dc:creator>
<dc:creator>Song, O.-R.</dc:creator>
<dc:creator>Broncel, M.</dc:creator>
<dc:creator>Russell, M.</dc:creator>
<dc:creator>Young, J. C.</dc:creator>
<dc:creator>Treeck, M.</dc:creator>
<dc:date>2022-03-21</dc:date>
<dc:identifier>doi:10.1101/2022.03.21.485090</dc:identifier>
<dc:title><![CDATA[Toxoplasma gondii ROP1 subverts murine and human innate immune restriction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.21.484343v1?rss=1">
<title>
<![CDATA[
Interacting active surfaces: a model for three-dimensional cell aggregates 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.21.484343v1?rss=1"
</link>
<description><![CDATA[
We introduce a modelling and simulation framework for cell aggregates in three dimensions based on interacting active surfaces. Cell mechanics is captured by a physical description of the acto-myosin cortex that includes cortical flows, viscous forces, active tensions, and bending moments. Cells interact with each other via short-range forces capturing the effect of adhesion molecules. We discretise the model constitutive equations using a finite element method, and provide a parallel implementation in C++. We discuss examples of application of this framework to simulations involving small and medium-sized aggregates: we consider the shape and dynamics of a cell doublet, a planar cell sheet, and a growing cell aggregate. This framework opens the door to the systematic exploration of the cell to tissue-scale mechanics of cell aggregates, which plays a key role in the morphogenesis of embryos and organoids.

Author summaryUnderstanding how tissue-scale morphogenesis arises from cell mechanics and cell-cell interactions is a fundamental question in developmental biology. Here we propose a mathematical and numerical framework to address this question. In this framework, each cell is described as an active surface representing the cell acto-myosin cortex, subjected to flows and shape changes according to active tensions, and to interaction with neighbouring cells in the tissue. Our method describes cellular processes such as cortical flows, cell adhesion, and cell shape changes in a deforming three-dimensional cellular aggregate. To solve the equations numerically, we employ a finite element discretisation, which allows us to solve for flows and cell shape changes with arbitrary resolution. We discuss applications of our framework to describe cell-cell adhesion in doublets, three-dimensional cell shape in a simple epithelium, and three-dimensional growth of a cell aggregate.
]]></description>
<dc:creator>Torres-Sanchez, A.</dc:creator>
<dc:creator>Winter, M. K.</dc:creator>
<dc:creator>Salbreux, G.</dc:creator>
<dc:date>2022-03-22</dc:date>
<dc:identifier>doi:10.1101/2022.03.21.484343</dc:identifier>
<dc:title><![CDATA[Interacting active surfaces: a model for three-dimensional cell aggregates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.22.485282v1?rss=1">
<title>
<![CDATA[
Occurrence and quantities of DNA modifications across the tree of life 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.22.485282v1?rss=1"
</link>
<description><![CDATA[
Enzymatic DNA modifications like methylcytosine (5mdC), methyladenine (N6mdA), or hydroxymethylcytosine (5hmdC) are key for chromatin function, gene expression regulation, and antiviral defense, but they remain understudied in non-model organisms. We established a mass spectrometric method for the sensitive and accurate quantification of enzymatic DNA modifications, and analyzed 85 bacterial genomes, 19 plant samples, 41 tissues from 12 animal species, 6 yeast species, and two archaeal species. We report no or only very low concentrations of DNA modifications in yeast and insects, but find DNA modifications universal to both bacteria and higher eukaryotes. Specifically for prokaryotes, our dataset indicates that evolutionary relationships and host-pathogen interactions, but not the ecological niche in general, select for a similar degree of DNA modification. In higher eukaryotes, largest concentration differences between tissues are detected for 5hmdC. Our dataset further reveals unique biological cases that warrant attention in the study of DNA modifications. For instance, while our data shows that most species contain just one dominating DNA modification, we detect all dominianting DNA modifications (5mdC, N6mdA, and 5hmdC) to exist in parallel in Raphanus sativus. Other plant species, like onion, sunflower, or the grass big bluestem, can have more than 35% of cytosines methylated. Finally, 5hmdC, so far mostly studied in the vertebrate central nervous system, was identified to reach a concentration of up to 8% of all cytosines in the Oman garra brain, and was also detected in several plants, like Lepidium sativum. The present study underscores the exploitation of biological diversity for studying DNA modifications.
]]></description>
<dc:creator>Varma, S. J.</dc:creator>
<dc:creator>Calvani, E.</dc:creator>
<dc:creator>Gruening, N.-M.</dc:creator>
<dc:creator>Messner, C. B.</dc:creator>
<dc:creator>Grayson, N.</dc:creator>
<dc:creator>Capuano, F.</dc:creator>
<dc:creator>Muelleder, M.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:date>2022-03-23</dc:date>
<dc:identifier>doi:10.1101/2022.03.22.485282</dc:identifier>
<dc:title><![CDATA[Occurrence and quantities of DNA modifications across the tree of life]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.26.485931v1?rss=1">
<title>
<![CDATA[
A Bayesian approach to incorporate structural data into the mapping of genotype to antigenic phenotype of influenza A(H3N2) viruses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.26.485931v1?rss=1"
</link>
<description><![CDATA[
Surface antigens of pathogens are commonly targeted by vaccine-elicited antibodies but antigenic variability, notably in RNA viruses such as influenza, HIV and SARS-CoV-2, pose challenges for control by vaccination. For example, influenza A(H3N2) entered the human population in 1968 causing a pandemic and has since been monitored, along with other seasonal influenza viruses, for the emergence of antigenic drift variants through intensive global surveillance and laboratory characterisation. Statistical models of the relationship between genetic differences among viruses and their antigenic similarity provide useful information to inform vaccine development though accurate identification of causative mutations is complicated by highly correlated genetic signals that arise due to the evolutionary process. Here, using a sparse hierarchical Bayesian analogue of an experimentally validated model for integrating genetic and antigenic data, we identify the genetic changes in influenza A(H3N2) virus that underpin antigenic drift. We show that incorporating protein structural data into variable selection helps resolve ambiguities arising due to correlated signals, with the proportion of variables representing haemagglutinin positions decisively included, or excluded, increased from 59.8% to 72.4%. The accuracy of variable selection judged by proximity to experimentally determined antigenic sites was improved simultaneously. Structure-guided variable selection thus improves confidence in the identification of genetic explanations of antigenic variation and we also show that prioritising the identification of causative mutations is not detrimental to the predictive capability of the analysis. Indeed, incorporating structural information into variable selection resulted in a model that could more accurately predict antigenic assay titres for phenotypically-uncharactrised virus from genetic sequence. Combined, these analyses have the potential to inform choices of reference viruses, the targeting of laboratory assays, and predictions of the evolutionary success of different genotypes, and can therefore be used to inform vaccine selection processes.
]]></description>
<dc:creator>Harvey, W. T.</dc:creator>
<dc:creator>Davies, V.</dc:creator>
<dc:creator>Daniels, R. S.</dc:creator>
<dc:creator>Whittaker, L.</dc:creator>
<dc:creator>Gregory, V.</dc:creator>
<dc:creator>Hay, A. J.</dc:creator>
<dc:creator>Husmeier, D.</dc:creator>
<dc:creator>McCauley, J. W.</dc:creator>
<dc:creator>Reeve, R.</dc:creator>
<dc:date>2022-03-27</dc:date>
<dc:identifier>doi:10.1101/2022.03.26.485931</dc:identifier>
<dc:title><![CDATA[A Bayesian approach to incorporate structural data into the mapping of genotype to antigenic phenotype of influenza A(H3N2) viruses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.28.486054v1?rss=1">
<title>
<![CDATA[
NDR1/2 kinases regulate membrane trafficking, enable efficient autophagy and prevent neurodegeneration 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.28.486054v1?rss=1"
</link>
<description><![CDATA[
Autophagy is essential for neuronal development and its deregulation contributes to neurodegenerative diseases. NDR1 and NDR2 are highly conserved kinases implicated in neuronal development, mitochondrial health and autophagy, but how they affect mammalian brain development in vivo is not known. Using single and double Ndr1/2 knockout mouse models we show that, dual, but not individual loss of Ndr1/2 in neurons causes neurodegeneration during brain development, but also in adult mice. Proteomic and phosphoproteomic comparisons between Ndr1/2 knockout and control brains revealed novel kinase substrates and indicated that endocytosis is significantly affected in the absence of NDR1/2. We validated the endocytic protein, Raph1/Lpd1 as a novel NDR1/2 substrate and showed that both NDR1/2 and Raph1 are critical for endocytosis and membrane recycling. In NDR1/2 knockout brains, we observed prominent accumulation of transferrin receptor, p62 and ubiquitinated proteins, indicative of a major impairment of protein homeostasis. Furthermore, the levels of LC3-positive autophagosomes were reduced in knockout neurons, implying that reduced autophagy efficiency mediates p62 accumulation and neurotoxicity. Mechanistically, pronounced mislocalisation of the transmembrane autophagy protein ATG9A at the neuronal periphery, impaired axonal ATG9A trafficking and increased ATG9A surface levels further confirm defects in membrane trafficking and could underlie the impairment in autophagy. We provide novel insight into the roles of NDR1/2 kinases in maintaining neuronal health.

HighlightsO_LIDual neuronal Ndr1 and Ndr2 knockout during development or in adult mice causes neurodegeneration.
C_LIO_LIPhosphoproteomics comparison of Ndr1/2 knockouts with control littermates shows endocytosis and membrane trafficking to be affected and reveals novel substrates.
C_LIO_LIRaph1/Lamellipodin is a novel NDR1/2 substrate that is required for TfR endocytosis.
C_LIO_LINdr1/2 knockout brains exhibit a severe defect in ubiquitinated protein clearance and reduced autophagy.
C_LIO_LINDR1/2 and Raph1 are required for the trafficking of the only transmembrane autophagy protein, ATG9A.
C_LI
]]></description>
<dc:creator>Ultanir, S. K.</dc:creator>
<dc:creator>Rosianu, F.</dc:creator>
<dc:creator>Mihaylov, S. R.</dc:creator>
<dc:creator>Eder, N.</dc:creator>
<dc:creator>Martiniuc, A.</dc:creator>
<dc:creator>Claxton, S.</dc:creator>
<dc:creator>Flynn, H.</dc:creator>
<dc:creator>Jalal, S.</dc:creator>
<dc:creator>Domart, M.-C.</dc:creator>
<dc:creator>Collinson, L. M.</dc:creator>
<dc:creator>Skehel, M.</dc:creator>
<dc:creator>Snijders, A. P. P.</dc:creator>
<dc:creator>Krause, M.</dc:creator>
<dc:creator>Tooze, S. A.</dc:creator>
<dc:date>2022-03-29</dc:date>
<dc:identifier>doi:10.1101/2022.03.28.486054</dc:identifier>
<dc:title><![CDATA[NDR1/2 kinases regulate membrane trafficking, enable efficient autophagy and prevent neurodegeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.06.487392v1?rss=1">
<title>
<![CDATA[
The natural diversity of the yeast proteome reveals chromosome-wide dosage compensation in aneuploids 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.06.487392v1?rss=1"
</link>
<description><![CDATA[
Aneuploidy, an imbalance in chromosome copy numbers, causes genetic disorders, and drives cancer progression, drug tolerance, and antimicrobial resistance. While aneuploidy can confer stress resistance, it is not well understood how cells overcome the fitness burden caused by aberrant chromosomal copy numbers. Studies using both systematically generated 1-5 and natural aneuploid yeasts 6-8 triggered an intense debate about the role of dosage compensation, concluding that aneuploidy is transmitted to the transcriptome and proteome without significant buffering at the chromosome-wide level, and is, at least in lab strains, associated with significant fitness costs. Conversely, systematic sequencing and phenotyping of large collections of natural isolates revealed that aneuploidy is frequent and has few - if any - fitness costs in nature 9. To address these discrepant findings at the proteomic level, we developed a platform that yields highly precise proteomic measurements across large numbers of genetically diverse samples, and applied it to natural isolates collected as part of the 1011 genomes project 9. For 613 of the isolates, we were able to match the proteomes to their corresponding transcriptomes and genomes, subsequently quantifying the effect of aneuploidy on gene expression by comparing 95 aneuploid with 518 euploid strains. We find, as in previous studies, that aneuploid gene dosage is not buffered chromosome-wide at the transcriptome level. Importantly, in the proteome, we detect an attenuation of aneuploidy by about 25% below the aneuploid gene dosage in natural yeast isolates. Furthermore, this chromosome-wide dosage compensation is associated with the ubiquitin-proteasome system (UPS), which is expressed at higher levels and has increased activity across natural aneuploid strains. Thus, through systematic exploration of the species-wide diversity of the yeast proteome, we shed light on a long-standing debate about the biology of aneuploids, revealing that aneuploidy tolerance is mediated through chromosome-wide dosage compensation at the proteome level.
]]></description>
<dc:creator>Muenzner, J.</dc:creator>
<dc:creator>Trebulle, P.</dc:creator>
<dc:creator>Agostini, F.</dc:creator>
<dc:creator>Messner, C. B.</dc:creator>
<dc:creator>Steger, M.</dc:creator>
<dc:creator>Lehmann, A.</dc:creator>
<dc:creator>Caudal, E.</dc:creator>
<dc:creator>Egger, A.-S.</dc:creator>
<dc:creator>Amari, F.</dc:creator>
<dc:creator>Barthel, N.</dc:creator>
<dc:creator>De Chiara, M.</dc:creator>
<dc:creator>Muelleder, M.</dc:creator>
<dc:creator>Demichev, V.</dc:creator>
<dc:creator>Liti, G.</dc:creator>
<dc:creator>Schacherer, J.</dc:creator>
<dc:creator>Gossmann, T.</dc:creator>
<dc:creator>Berman, J.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:date>2022-04-08</dc:date>
<dc:identifier>doi:10.1101/2022.04.06.487392</dc:identifier>
<dc:title><![CDATA[The natural diversity of the yeast proteome reveals chromosome-wide dosage compensation in aneuploids]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.08.487575v1?rss=1">
<title>
<![CDATA[
Phosphorylation of the novel mTOR substrate Unkempt regulates cellular morphogenesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.08.487575v1?rss=1"
</link>
<description><![CDATA[
Mechanistic target of rapamycin (mTOR) is a protein kinase that integrates multiple inputs to regulate anabolic cellular processes. mTOR complex I (mTORC1) has key functions in growth control, autophagy and metabolism. Much less is known about the signalling components that act downstream of mTORC1 that regulate cellular morphology, a vital determinant of cellular function. Here we show that the RNA-binding protein Unkempt, a key regulator of cellular morphogenesis, is a novel substrate mTORC1. We find that Unkempt phosphorylation is regulated by nutrient levels and growth factors via mTORC1. Furthermore, Unkempt physically interacts with and is directly phosphorylated by mTORC1 through binding to the regulatory-associated protein of mTOR, Raptor. Phosphorylation of Unkempt, which we find is mTORC1-dependent in cultured mammalian cell lines as well as in primary tissues, occurs largely within the highly serine-rich intrinsically disordered region of Unkempt. Importantly, mutation analysis of this region indicates that phosphorylation inhibits the ability of Unkempt to induce a bipolar morphology. Our findings reveal a novel molecular link between mTORC1 signalling and cellular morphogenesis.
]]></description>
<dc:creator>Baskaran, P.</dc:creator>
<dc:creator>Mihaylov, S. R.</dc:creator>
<dc:creator>Vinsland, E.</dc:creator>
<dc:creator>Shah, K.</dc:creator>
<dc:creator>Granat, L.</dc:creator>
<dc:creator>Ultanir, S. K.</dc:creator>
<dc:creator>Tee, A. R.</dc:creator>
<dc:creator>Murn, J.</dc:creator>
<dc:creator>Bateman, J. M.</dc:creator>
<dc:date>2022-04-10</dc:date>
<dc:identifier>doi:10.1101/2022.04.08.487575</dc:identifier>
<dc:title><![CDATA[Phosphorylation of the novel mTOR substrate Unkempt regulates cellular morphogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.07.487490v1?rss=1">
<title>
<![CDATA[
High-fidelity 3D live-cell nanoscopy through data-driven enhanced super-resolution radial fluctuation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.07.487490v1?rss=1"
</link>
<description><![CDATA[
In recent years, the development of new image analysis approaches has highlighted the possibility of recovering superresolution information from short sequences of wide-field images. Our recently developed method, SRRF (Super-Resolution Radial Fluctuations), enables long-term live-cell imaging beyond the resolution limit without specialized hardware. Here, we present eSRRF (enhanced-SRRF), a significant improvement over our initial method, enhancing image fidelity to the underlying structure and resolution. Especially, eSRRF uses automated data-driven parameter optimization, including an estimation of the number of frames necessary for optimal reconstruction. We demonstrate the improved fidelity of the images reconstructed with eSRRF and highlight its versatility and ease of use over a wide range of microscopy techniques and biological systems. We also extend eSRRF to 3D super-resolution microscopy by combining it with multi-focus microscopy (MFM), obtaining volumetric super-resolution imaging of live cells with acquisition speed of ~1 volume/second.
]]></description>
<dc:creator>Laine, R. F.</dc:creator>
<dc:creator>Heil, H. S.</dc:creator>
<dc:creator>Coelho, S.</dc:creator>
<dc:creator>Nixon-Abell, J.</dc:creator>
<dc:creator>Jimenez, A.</dc:creator>
<dc:creator>Galgani, T.</dc:creator>
<dc:creator>Stubb, A.</dc:creator>
<dc:creator>Follain, G.</dc:creator>
<dc:creator>Culley, S.</dc:creator>
<dc:creator>Jacquemet, G.</dc:creator>
<dc:creator>Hajj, B.</dc:creator>
<dc:creator>Leterrier, C.</dc:creator>
<dc:creator>Henriques, R.</dc:creator>
<dc:date>2022-04-08</dc:date>
<dc:identifier>doi:10.1101/2022.04.07.487490</dc:identifier>
<dc:title><![CDATA[High-fidelity 3D live-cell nanoscopy through data-driven enhanced super-resolution radial fluctuation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.13.488127v1?rss=1">
<title>
<![CDATA[
A quantitative and spatial analysis of cell cycle regulators during the fission yeast cell cycle 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.13.488127v1?rss=1"
</link>
<description><![CDATA[
We have carried out a systems-level analysis of the spatial and temporal dynamics of cell cycle regulators in the fission yeast Schizosaccharomyces pombe. In a comprehensive single cell analysis we have precisely quantified the levels of 38 proteins previously identified as regulators of the G2 to mitosis transition, and of 7 proteins acting at the G1 to S-phase transition. Only two of the 38 mitotic regulators exhibit changes in concentration at the whole cell level, the mitotic B-type cyclin Cdc13 which accumulates continually throughout the cell cycle, and the regulatory phosphatase Cdc25 which exhibits a complex cell cycle pattern. Both proteins show similar patterns of change within the nucleus as in the whole cell but at higher concentrations. In addition, the concentrations of the major fission yeast cyclin dependent kinase (CDK) Cdc2, the CDK regulator Suc1 and the inhibitory kinase Wee1 also increase in the nucleus peaking at mitotic onset but are constant in the whole cell. The significant increase in concentration with size for Cdc13 supports the model that mitotic B-type cyclin accumulation acts as a cell size sensor. We propose a two-step process for the control of mitosis. First, Cdc13 accumulates in a size-dependent manner which drives increasing CDK activity. Second, from mid G2 the increasing nuclear accumulation of Cdc25 and the counteracting Wee1 introduces a bistability switch that results in a rapid rise of CDK activity at the end of G2 and thus brings about an orderly progression into mitosis.

Significance StatementAcross eukaryotes the increasing level of cyclin dependent kinase (CDK) activity drives progression through the cell cycle. As most cells divide at specific sizes, information responding to the size of the cell must feed into the regulation of CDK activity. In this study, we use fission yeast to precisely measure how proteins that have been previously identified in genome wide screens as cell cycle regulators change in their levels with cell cycle progression. We identify the mitotic B-type cyclin Cdc13 and mitotic inhibitory phosphatase Cdc25 as the only two proteins that change in both whole cell and nuclear concentration through the cell cycle, making them candidates for universal cell size sensors at the onset of mitosis and cell division.
]]></description>
<dc:creator>Curran, S.</dc:creator>
<dc:creator>Dey, G.</dc:creator>
<dc:creator>Rees, P.</dc:creator>
<dc:creator>Nurse, P.</dc:creator>
<dc:date>2022-04-13</dc:date>
<dc:identifier>doi:10.1101/2022.04.13.488127</dc:identifier>
<dc:title><![CDATA[A quantitative and spatial analysis of cell cycle regulators during the fission yeast cell cycle]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.13.488173v1?rss=1">
<title>
<![CDATA[
Low-efficiency conversion of proliferative glia into induced neurons by Ascl1 in the postnatal mouse cerebral cortex in vivo 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.13.488173v1?rss=1"
</link>
<description><![CDATA[
The proneural transcription factor Achaete-scute complex-like 1 (Ascl1) is a major regulator of neural progenitor fate, implicated both in neurogenesis and oligodendrogliogenesis. Ascl1 has been widely used to reprogram non-neuronal cells into induced neurons. In vitro, Ascl1 induces efficient reprogramming of proliferative astroglia from the early postnatal cerebral cortex into interneuron-like cells. Here, we examined whether Ascl1 can similarly induce neuronal reprogramming of glia undergoing proliferation in the postnatal mouse cerebral cortex in vivo. Toward this, we targeted cortical glia at the peak of proliferative expansion (i.e., postnatal day 5) by injecting a retrovirus encoding for Ascl1 into the mouse cerebral cortex. In sharp contrast to the very efficient reprogramming in vitro, Ascl1-transduced glial cells were converted into doublecortin-immunoreactive neurons only with low efficiency in vivo. Interfering with the phosphorylation of Ascl1 by mutation of six conserved proline-directed serine/threonine phosphorylation sites (Ascl1SA6) has been previously shown to increase its neurogenic activity in the early embryonic cerebral cortex. We therefore tested whether transduction of proliferative glia with a retrovirus encoding Ascl1SA6 improved their conversion into neurons. While in vitro glia-to-neuron conversion was markedly enhanced, in vivo reprogramming efficiency remained low. However, both wild-type and mutant Ascl1 reduced the relative number of cells expressing the astrocytic marker glial fibrillary acidic protein (GFAP) and increased the relative number of cells expressing the oligodendroglial marker Sox10 in vivo. Together, our results indicate that the enhanced neurogenic response of proliferative postnatal glia to Ascl1SA6 versus Ascl1 observed in vitro is not recapitulated in vivo.
]]></description>
<dc:creator>Galante, C.</dc:creator>
<dc:creator>Marichal, N.</dc:creator>
<dc:creator>Schuurmans, C.</dc:creator>
<dc:creator>Berninger, B.</dc:creator>
<dc:creator>Peron, S.</dc:creator>
<dc:date>2022-04-13</dc:date>
<dc:identifier>doi:10.1101/2022.04.13.488173</dc:identifier>
<dc:title><![CDATA[Low-efficiency conversion of proliferative glia into induced neurons by Ascl1 in the postnatal mouse cerebral cortex in vivo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.11.487796v1?rss=1">
<title>
<![CDATA[
Multimodal single cell data integration challenge: results and lessons learned 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.11.487796v1?rss=1"
</link>
<description><![CDATA[
Biology has become a data-intensive science. Recent technological advances in single-cell genomics have enabled the measurement of multiple facets of cellular state, producing datasets with millions of single-cell observations. While these data hold great promise for understanding molecular mechanisms in health and disease, analysis challenges arising from sparsity, technical and biological variability, and high dimensionality of the data hinder the derivation of such mechanistic insights. To promote the innovation of algorithms for analysis of multimodal single-cell data, we organized a competition at NeurIPS 2021 applying the Common Task Framework to multimodal single-cell data integration. For this competition we generated the first multimodal benchmarking dataset for single-cell biology and defined three tasks in this domain: prediction of missing modalities, aligning modalities, and learning a joint representation across modalities. We further specified evaluation metrics and developed a cloud-based algorithm evaluation pipeline. Using this setup, 280 competitors submitted over 2600 proposed solutions within a 3 month period, showcasing substantial innovation especially in the modality alignment task. Here, we present the results, describe trends of well performing approaches, and discuss challenges associated with running the competition.
]]></description>
<dc:creator>Lance, C.</dc:creator>
<dc:creator>Luecken, M. D.</dc:creator>
<dc:creator>Burkhardt, D. B.</dc:creator>
<dc:creator>Cannoodt, R.</dc:creator>
<dc:creator>Rautenstrauch, P.</dc:creator>
<dc:creator>Laddach, A. C.</dc:creator>
<dc:creator>Ubingazhibov, A.</dc:creator>
<dc:creator>Cao, Z.-J.</dc:creator>
<dc:creator>Deng, K.</dc:creator>
<dc:creator>Khan, S.</dc:creator>
<dc:creator>Liu, Q.</dc:creator>
<dc:creator>Russkikh, N.</dc:creator>
<dc:creator>Ryazantsev, G.</dc:creator>
<dc:creator>Ohler, U.</dc:creator>
<dc:creator>NeurIPS 2021 Multimodal data integration competition participants,</dc:creator>
<dc:creator>Pisco, A. O.</dc:creator>
<dc:creator>Bloom, J. M.</dc:creator>
<dc:creator>Krishnaswamy, S.</dc:creator>
<dc:creator>Theis, F. J.</dc:creator>
<dc:date>2022-04-12</dc:date>
<dc:identifier>doi:10.1101/2022.04.11.487796</dc:identifier>
<dc:title><![CDATA[Multimodal single cell data integration challenge: results and lessons learned]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.14.488299v1?rss=1">
<title>
<![CDATA[
High-throughput proteomics of nanogram-scale samples with Zeno SWATH DIA 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.14.488299v1?rss=1"
</link>
<description><![CDATA[
The ability to conduct high-quality proteomic experiments in high throughput has opened new avenues in clinical research, drug discovery, and systems biology. Next to an increase in quantitative precision, recent developments in high-throughput proteomics have also gained proteomic depth, to the extent that earlier gaps between classic and high-throughput experiments have significantly narrowed. Here we introduce and benchmark Zeno SWATH, a data-independent acquisition technique that employs a linear ion trap pulsing (Zeno trap pulsing) in order to increase proteomic depth and dynamic range in proteomic experiments. Combined with the high acquisition speed, these gains in sensitivity are particularly attractive for conducting high-throughput proteomics experiments with high chromatographic flow rates and fast gradients. We demonstrate that when combined with either micro-flow- or analytical-flow-rate chromatography, Zeno SWATH increases protein identification in complex samples 5- to 10-fold when compared to current SWATH acquisition methods on the same instrument. Using 20-min micro-flow chromatography, Zeno SWATH identified > 6,000 proteins from a 62.5 ng load of human cell lysate with more than 5,000 proteins consistently quantified in triplicate injections with a median CV of 6%. Using 5-min analytical-flow-rate chromatography (800 {micro}l/min), Zeno SWATH identified 4,907 proteins from a triplicate injection of 2 {micro}g of a human cell lysate; or more than 3,000 proteins from 250 ng tryptic digest. Zeno SWATH hence facilitates precise proteomic experiments with small sample amounts using a fast and robust high flow-rate chromatographic method, broadening the application space that requires precise proteomic experiments on a large scale.
]]></description>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Muelleder, M.</dc:creator>
<dc:creator>Batruch, I.</dc:creator>
<dc:creator>Chelur, A.</dc:creator>
<dc:creator>Textoris-Taube, K.</dc:creator>
<dc:creator>Schwecke, T.</dc:creator>
<dc:creator>Hartl, J.</dc:creator>
<dc:creator>Causon, J.</dc:creator>
<dc:creator>Castro-Perez, J.</dc:creator>
<dc:creator>Demichev, V.</dc:creator>
<dc:creator>Tate, S.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:date>2022-04-14</dc:date>
<dc:identifier>doi:10.1101/2022.04.14.488299</dc:identifier>
<dc:title><![CDATA[High-throughput proteomics of nanogram-scale samples with Zeno SWATH DIA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.15.488437v1?rss=1">
<title>
<![CDATA[
Kinesin-1 transports morphologically distinct intracellular virions during vaccinia infection 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.15.488437v1?rss=1"
</link>
<description><![CDATA[
Intracellular mature virions (IMV) are the first and most abundant infectious form of vaccinia virus to assemble during its replication cycle. IMV can undergo microtubule-based motility, but their directionality and the motor involved in their transport remain unknown. Here, we demonstrate that IMV, like intracellular enveloped virions (IEV), the second form of vaccinia, undergo anterograde transport and recruit kinesin-1. In vitro reconstitution of virion transport reveals that IMV and IEV move toward microtubule plus-ends with respective velocities of 0.66 and 0.56 m/s. Quantitative imaging establishes IMV and IEV recruit an average of 65 and 115 kinesin-1 motor complexes respectively. In the absence of kinesin-1 there is a near-complete loss of in vitro motility and defects in the cellular spread of both virions. Our observations demonstrate kinesin-1 transports two morphologically distinct forms of vaccinia. Reconstitution of vaccinia-based microtubule motility in vitro provides a new model to investigate how motor number and regulation impacts transport of a bona fide kinesin-1 cargo.
]]></description>
<dc:creator>Xu, A.</dc:creator>
<dc:creator>Basant, A.</dc:creator>
<dc:creator>Schleich, S.</dc:creator>
<dc:creator>Newsome, T. P.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:date>2022-04-15</dc:date>
<dc:identifier>doi:10.1101/2022.04.15.488437</dc:identifier>
<dc:title><![CDATA[Kinesin-1 transports morphologically distinct intracellular virions during vaccinia infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.14.488310v1?rss=1">
<title>
<![CDATA[
Arousal neurons that anticipate deviations in blood glucose. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.14.488310v1?rss=1"
</link>
<description><![CDATA[
Blood glucose variability shapes human brain performance and diverse clinical outcomes. However, it remains poorly understood how blood glucose fluctuations are decoded by genetically-defined neurons to change brain activity and behavior. Recent breakthroughs in genetics and clinical diagnostics identified hypothalamic hypocretin/orexin neurons (HONs) as core determinants of brain activity and adaptive behavior across mammals. Here we show that low-frequency HON population waves are tuned for transmitting information about minute-to-minute temporal features of blood glucose, thus rapidly converting its variability into brain state of behaving mice. Contrary to current theories envisioning glucose-proportional neural responses, the HONs response tracked blood glucose gradients, thus generating efficient neural adaptations in anticipation of maximal glucose deviations. Resolving this population response at the single cell level with volumetric multiphoton imaging furthermore revealed glucose-excited and glucose-inhibited HONs, distinctly coupled to body movements in the high-frequency domain. Finally, HON-selective opotogenetics and cell ablation demonstrated that HONs are critical for linking glucose to adaptive behavior. These results provide an insight into how behaviorally influential hypothalamic networks interpret blood glucose variability. This may inform future metrics for efficient prediction of glycemic states in health and disease.
]]></description>
<dc:creator>Viskaitis, P.</dc:creator>
<dc:creator>Tesmer, A.</dc:creator>
<dc:creator>Karnani, M. M.</dc:creator>
<dc:creator>Arnold, M.</dc:creator>
<dc:creator>Donegan, D.</dc:creator>
<dc:creator>Bracey, E.</dc:creator>
<dc:creator>Peleg-Raibstein, D.</dc:creator>
<dc:creator>Burdakov, D.</dc:creator>
<dc:date>2022-04-15</dc:date>
<dc:identifier>doi:10.1101/2022.04.14.488310</dc:identifier>
<dc:title><![CDATA[Arousal neurons that anticipate deviations in blood glucose.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.18.488654v1?rss=1">
<title>
<![CDATA[
The DNA replication initiation protein DnaD is recruited to a specific strand of the Bacillus subtilis chromosome origin 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.18.488654v1?rss=1"
</link>
<description><![CDATA[
Genome replication is a fundamental biological activity shared by all organisms. Chromosomal replication proceeds bidirectionally from origins, requiring the loading of two helicases, one for each replisome. The molecular mechanisms for helicase loading at bacterial chromosome origins (oriC) are unclear. Here we investigated the essential DNA replication initiation protein DnaD in the model organism Bacillus subtilis. A set of DnaD residues required for ssDNA binding was identified, and photo-crosslinking revealed that this ssDNA binding region interacts preferentially with one strand of oriC. Biochemical and genetic data support the model that DnaD recognizes a new single-stranded DNA (ssDNA) motif located in oriC (DnaD Recognition Element, "DRE"). Considered with cryo-electron microscopy (cryo-EM) imaging of full length DnaD, we propose that the location of the DRE within the oriC orchestrates strand-specific recruitment of helicase to achieve bidirectional DNA replication. These findings significantly advance our mechanistic understanding of bidirectional replication from a bacterial chromosome origin.
]]></description>
<dc:creator>Winterhalter, C.</dc:creator>
<dc:creator>Pelliciari, S.</dc:creator>
<dc:creator>Stevens, D.</dc:creator>
<dc:creator>Fenyk, S.</dc:creator>
<dc:creator>Marchand, E.</dc:creator>
<dc:creator>Cronin, N.</dc:creator>
<dc:creator>Soultanas, P.</dc:creator>
<dc:creator>Costa, T.</dc:creator>
<dc:creator>Ilangovan, A.</dc:creator>
<dc:creator>Murray, H.</dc:creator>
<dc:date>2022-04-18</dc:date>
<dc:identifier>doi:10.1101/2022.04.18.488654</dc:identifier>
<dc:title><![CDATA[The DNA replication initiation protein DnaD is recruited to a specific strand of the Bacillus subtilis chromosome origin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.20.488895v1?rss=1">
<title>
<![CDATA[
Emergence of new subgenomic mRNAs in SARS-CoV-2 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.20.488895v1?rss=1"
</link>
<description><![CDATA[
Two mutations occurred in SARS-CoV-2 early during the COVID-19 pandemic that have come to define circulating virus lineages1: first a change in the spike protein (D614G) that defines the B.1 lineage and second, a double substitution in the nucleocapsid protein (R203K, G204R) that defines the B.1.1 lineage, which has subsequently given rise to three Variants of Concern: Alpha, Gamma and Omicron. While the latter mutations appear unremarkable at the protein level, there are dramatic implications at the nucleotide level: the GGG[-&gt;]AAC substitution generates a new Transcription Regulatory Sequence (TRS) motif, driving SARS-CoV-2 to express a novel subgenomic mRNA (sgmRNA) encoding a truncated C-terminal portion of nucleocapsid (N.iORF3), which is an inhibitor of type I interferon production. We find that N.iORF3 also emerged independently within the Iota variant, and further show that additional TRS motifs have convergently evolved to express novel sgmRNAs; notably upstream of Spike within the nsp16 coding region of ORF1b, which is expressed during human infection. Our findings demonstrate that SARS-CoV-2 is undergoing evolutionary changes at the functional RNA level in addition to the amino acid level, reminiscent of eukaryotic evolution. Greater attention to this aspect in the assessment of emerging strains of SARS-CoV-2 is warranted.
]]></description>
<dc:creator>Mears, H. V.</dc:creator>
<dc:creator>Young, G. R.</dc:creator>
<dc:creator>Sanderson, T.</dc:creator>
<dc:creator>Harvey, R.</dc:creator>
<dc:creator>Crawford, M.</dc:creator>
<dc:creator>Snell, D. M.</dc:creator>
<dc:creator>Fowler, A. S.</dc:creator>
<dc:creator>Hussain, S.</dc:creator>
<dc:creator>Nicod, J.</dc:creator>
<dc:creator>Peacock, T. P.</dc:creator>
<dc:creator>Emmott, E.</dc:creator>
<dc:creator>Finsterbusch, K.</dc:creator>
<dc:creator>Luptak, J.</dc:creator>
<dc:creator>Wall, E.</dc:creator>
<dc:creator>Williams, B.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Bauer, D. L.</dc:creator>
<dc:date>2022-04-20</dc:date>
<dc:identifier>doi:10.1101/2022.04.20.488895</dc:identifier>
<dc:title><![CDATA[Emergence of new subgenomic mRNAs in SARS-CoV-2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.22.488007v1?rss=1">
<title>
<![CDATA[
Crosslinker design determines microtubule network organization by opposing motors 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.22.488007v1?rss=1"
</link>
<description><![CDATA[
During cell division, crosslinking motors determine the architecture of the spindle, a dynamic microtubule network that segregates the chromosomes. It is unclear how motors with opposite directionality coordinate to drive both contractile and extensile behaviors in the spindle. Particularly, the impact of different crosslinker designs on network self-organization is not understood, limiting our understanding of self-organizing structures in cells, but also our ability to engineer new active materials. Here, we use experiment and theory to examine active microtubule networks driven by mixtures of motors with opposite directionality and different crosslinker design. We find that although the kinesin-14 HSET causes network contraction when dominant, it can also assist the opposing kinesin-5 KIF11 to generate extensile networks. This bifunctionality results from HSETs asymmetric design, distinct from symmetric KIF11. These findings expand the set of rules underlying patterning of active microtubule assemblies and allow a better understanding of motor cooperation in the spindle.

SIGNIFICANCE STATEMENTDuring cell division, the spindle apparatus segregates duplicated chromosomes for their inheritance by the daughter cells. The spindle is a highly interconnected network of microtubule filaments that are crosslinked by different types of molecular motors. How the different motors cooperate to organize the spindle network is not understood. Here, we show that an asymmetric crosslinker design can confer bifunctionality to a mitotic motor in the presence of other motors. The asymmetric motor supports both extensile and contractile microtubule network behaviors as observed in different parts of the spindle. These findings define new rules controlling the generation of active microtubule networks and allow us to better understand how motors cooperate to organize the correct spindle architecture when a cell divides.
]]></description>
<dc:creator>Henkin, G.</dc:creator>
<dc:creator>Chew, W.-X.</dc:creator>
<dc:creator>Nedelec, F.</dc:creator>
<dc:creator>Surrey, T.</dc:creator>
<dc:date>2022-04-22</dc:date>
<dc:identifier>doi:10.1101/2022.04.22.488007</dc:identifier>
<dc:title><![CDATA[Crosslinker design determines microtubule network organization by opposing motors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.22.489156v1?rss=1">
<title>
<![CDATA[
Nodal signaling establishes a competency window for stochastic cell fate switching 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.22.489156v1?rss=1"
</link>
<description><![CDATA[
Specification of the germ layers by Nodal signaling has long been regarded as an archetype of how graded morphogens induce different cell fates. However, this deterministic model cannot explain why only a subset of cells at the margin of the early zebrafish embryo adopt the endodermal fate, while their immediate neighbours, experiencing similar signaling profiles, become mesoderm. Combining pharmacology, quantitative imaging and single cell transcriptomics, we demonstrate that sustained Nodal signaling establishes a bipotential progenitor state where cells initially fated to become mesoderm can switch to an endodermal fate. Switching is a random event, the likelihood of which is modulated by Fgf signaling. This inherently imprecise mechanism nevertheless leads to robust endoderm formation because of buffering at later stages. Thus, in contrast to previous deterministic models of morphogen action, Nodal establishes a temporal window when cells are competent to undergo a stochastic cell fate switch, rather than determining fate itself.
]]></description>
<dc:creator>Economou, A. D.</dc:creator>
<dc:creator>Guglielmi, L.</dc:creator>
<dc:creator>East, P.</dc:creator>
<dc:creator>Hill, C. S.</dc:creator>
<dc:date>2022-04-22</dc:date>
<dc:identifier>doi:10.1101/2022.04.22.489156</dc:identifier>
<dc:title><![CDATA[Nodal signaling establishes a competency window for stochastic cell fate switching]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.21.489072v1?rss=1">
<title>
<![CDATA[
Impaired immune response drives age-dependent severity of COVID-19 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.21.489072v1?rss=1"
</link>
<description><![CDATA[
SARS-CoV-2 is a highly contagious respiratory virus and the causative agent for COVID-19. The severity of disease varies from mildly symptomatic to lethal and shows an extraordinary correlation with increasing age, which represents the major risk factor for severe COVID-191. However, the precise pathomechanisms leading to aggravated disease in the elderly are currently unknown. Delayed and insufficient antiviral immune responses early after infection as well as dysregulated and overshooting immunopathological processes late during disease were suggested as possible mechanisms. Here we show that the age-dependent increase of COVID-19 severity is caused by the disruption of a timely and well-coordinated innate and adaptive immune response due to impaired interferon (IFN) responses. To overcome the limitations of mechanistic studies in humans, we generated a mouse model for severe COVID-19 and compared the kinetics of the immune responses in adult and aged mice at different time points after infection. Aggravated disease in aged mice was characterized by a diminished IFN-{gamma} response and excessive virus replication. Accordingly, adult IFN-{gamma} receptor-deficient mice phenocopied the age-related disease severity and supplementation of IFN-{gamma} reversed the increased disease susceptibility of aged mice.

Mimicking impaired type I IFN immunity in adult and aged mice, a second major risk factor for severe COVID-192-4, we found that therapeutic treatment with IFN-{lambda} in adult and a combinatorial treatment with IFN-{gamma} and IFN-{lambda} in aged Ifnar1-/-mice was highly efficient in protecting against severe disease.

Our findings provide an explanation for the age-dependent disease severity of COVID-19 and clarify the nonredundant antiviral functions of type I, II and III IFNs during SARS-CoV-2 infection in an age-dependent manner. Based on our data, we suggest that highly vulnerable individuals combining both risk factors, advanced age and an impaired type I IFN immunity, may greatly benefit from immunotherapy combining IFN-{gamma} and IFN-{lambda}.
]]></description>
<dc:creator>Beer, J.</dc:creator>
<dc:creator>Crotta, S.</dc:creator>
<dc:creator>Breithaupt, A.</dc:creator>
<dc:creator>Ohnemus, A.</dc:creator>
<dc:creator>Becker, J.</dc:creator>
<dc:creator>Sachs, B.</dc:creator>
<dc:creator>Kern, L.</dc:creator>
<dc:creator>Llorian, M.</dc:creator>
<dc:creator>Ebert, N.</dc:creator>
<dc:creator>Labroussaa, F.</dc:creator>
<dc:creator>Tran, T. T. N.</dc:creator>
<dc:creator>Trueeb, B. S.</dc:creator>
<dc:creator>Jores, J.</dc:creator>
<dc:creator>Thiel, V.</dc:creator>
<dc:creator>Beer, M.</dc:creator>
<dc:creator>Fuchs, J.</dc:creator>
<dc:creator>Kochs, G.</dc:creator>
<dc:creator>Wack, A.</dc:creator>
<dc:creator>Schwemmle, M.</dc:creator>
<dc:creator>Schnepf, D.</dc:creator>
<dc:date>2022-04-22</dc:date>
<dc:identifier>doi:10.1101/2022.04.21.489072</dc:identifier>
<dc:title><![CDATA[Impaired immune response drives age-dependent severity of COVID-19]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.26.489550v1?rss=1">
<title>
<![CDATA[
Bimodal regulation of axonal transport by the GDNF-RET signalling axis in healthy and diseased motor neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.26.489550v1?rss=1"
</link>
<description><![CDATA[
Deficits in axonal transport are one of the earliest pathological outcomes in several models of amyotrophic lateral sclerosis (ALS), including SOD1G93A mice. Evidence suggests that rescuing these deficits prevents disease progression, stops denervation, and extends survival. Kinase inhibitors have been previously identified as transport enhancers, and are being investigated as potential therapies for ALS. For example, inhibitors of p38 mitogen-activated protein kinase and insulin growth factor receptor 1 have been shown to rescue axonal transport deficits in vivo in symptomatic SOD1G93A mice. In this work, we investigated the impact of RET, the tyrosine kinase receptor for glial cell-line-derived neurotrophic factor (GDNF), as a modifier of axonal transport. We identified fundamental interplay between RET signalling and axonal transport in both wild type and SOD1G93A motor neurons in vitro. We demonstrated that blockade of RET signalling using pharmacological inhibitors and genetic knockdown enhances signalling endosome transport in wild type motor neurons and uncovered a divergence in the response of primary motor neurons to GDNF compared with cell lines. Finally, we demonstrated that inhibition of the GDNF-RET signalling axis rescues in vivo transport deficits in early symptomatic SOD1G93A mice, promoting RET as a potential therapeutic target in the treatment of ALS.
]]></description>
<dc:creator>Rhymes, E. R.</dc:creator>
<dc:creator>Tosolini, A. P.</dc:creator>
<dc:creator>Fellows, A. D.</dc:creator>
<dc:creator>Mahy, W.</dc:creator>
<dc:creator>McDonald, N. Q.</dc:creator>
<dc:creator>Schiavo, G.</dc:creator>
<dc:date>2022-04-27</dc:date>
<dc:identifier>doi:10.1101/2022.04.26.489550</dc:identifier>
<dc:title><![CDATA[Bimodal regulation of axonal transport by the GDNF-RET signalling axis in healthy and diseased motor neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.29.488986v1?rss=1">
<title>
<![CDATA[
H4K16ac activates transcription of transposable elements and contribute to their cis regulatory function 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.29.488986v1?rss=1"
</link>
<description><![CDATA[
Mammalian genomes harbour a large number of transposable elements (TEs) and their remnants. Many epigenetic repression mechanisms are known to silence TE transcription. However, TEs are upregulated during early development, neuronal lineage, and cancers, although the epigenetic factors contributing to the transcription of TEs have yet to be fully elucidated. Here we demonstrated that the male-specific lethal (MSL) complex mediated acetylation of histone H4 lysine 16 (H4K16ac) activates transcription of long interspersed nuclear elements (LINE1, L1) and long terminal repeats (LTRs). Furthermore, we show that the H4K16ac marked L1 and LTR subfamilies function as enhancers and are enriched with chromatin features associated with active enhancers and looping factors. L1 and LTRs enriched with histone acetylations are bound by chromatin looping factors and these regions loop with genes. CRISPR-based epigenetic perturbation and genetic deletion of L1s reveal that H4K16ac marked L1s and LTRs regulate the expression of genes in cis. Overall, TEs enriched with H4K16ac contribute to the cis-regulatory landscape of a significant portion of the mammalian genome by maintaining an active chromatin landscape at TEs.

One Sentence SummaryH4K16ac activates LINE1 and ERV/LTR transcription and rewires the cis-regulatory landscape of a significant portion of the mammalian genome by increasing the transcriptional activity at TEs.
]]></description>
<dc:creator>Pal, D.</dc:creator>
<dc:creator>Patel, M.</dc:creator>
<dc:creator>Boulet, F.</dc:creator>
<dc:creator>Sundarraj, J.</dc:creator>
<dc:creator>Grant, O. A.</dc:creator>
<dc:creator>Branco, M. R.</dc:creator>
<dc:creator>Basu, S.</dc:creator>
<dc:creator>Zabet, N. R.</dc:creator>
<dc:creator>Scaffidi, P.</dc:creator>
<dc:creator>Pradeepa, M. M.</dc:creator>
<dc:date>2022-04-30</dc:date>
<dc:identifier>doi:10.1101/2022.04.29.488986</dc:identifier>
<dc:title><![CDATA[H4K16ac activates transcription of transposable elements and contribute to their cis regulatory function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.05.01.490232v1?rss=1">
<title>
<![CDATA[
PfHDAC1 is an essential regulator of parasite asexual growth with its altered genomic occupancy and activity associated with artemisinin drug resistance in Plasmodium falciparum 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.05.01.490232v1?rss=1"
</link>
<description><![CDATA[
Plasmodium falciparum is a deadly protozoan parasite and the causative agent of malaria, which accounts for close to 200 million cases and 400,000 deaths every year. It has been identified to possess a tightly regulated gene expression profile that is integrally linked to its timely development during the intraerythrocytic stage. Epigenetic modifiers of the histone acetylation code have been identified as key regulators of the parasites transcriptome. In this study, we characterize the solitary class I histone deacetylase PfHDAC1 and demonstrate that phosphorylation is required for its catalytic activity. PfHDAC1 binds to and regulates parasite genes responsible for housekeeping and stress-responsive functions. We show that PfHDAC1 activity in parasites is crucial for normal intraerythrocytic development and that its cellular abundance is correlated with parasitemia progression. We further show that PfHDAC1 has differential abundance and genomic occupancy in artemisinin drug-resistant vs sensitive parasites and that inhibition of its deacetylase activity can modulate the sensitivity of parasites to the drug. We also identify that artemisinin exposure can interfere with PfHDAC1 phosphorylation and its genomic occupancy. Collectively, our results demonstrate PfHDAC1 to be an important regulator of basic biological functions in parasites while also deterministic of responses to environmental stresses such as antimalarial drugs.
]]></description>
<dc:creator>Kanyal, A.</dc:creator>
<dc:creator>Davies, H.</dc:creator>
<dc:creator>Deshmukh, B.</dc:creator>
<dc:creator>Farheen, D.</dc:creator>
<dc:creator>Treeck, M.</dc:creator>
<dc:creator>Karmodiya, K.</dc:creator>
<dc:date>2022-05-02</dc:date>
<dc:identifier>doi:10.1101/2022.05.01.490232</dc:identifier>
<dc:title><![CDATA[PfHDAC1 is an essential regulator of parasite asexual growth with its altered genomic occupancy and activity associated with artemisinin drug resistance in Plasmodium falciparum]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.05.03.490423v1?rss=1">
<title>
<![CDATA[
Digital holography-based 3D particle localisation for single molecule tweezer techniques 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.05.03.490423v1?rss=1"
</link>
<description><![CDATA[
We present a three-dimensional imaging technique for fast tracking of microscopic objects in a fluid environment. Our technique couples digital holographic microscopy with three-dimensional localisation via parabolic masking. Compared with existing approaches, our method reconstructs 3D volumes from single-plane images, which greatly simplifies image acquisition, reduces the demand on microscope hardware, and facilitates tracking higher densities of microscopic particles while maintaining similar levels of precision. We demonstrate utility of this method in magnetic tweezer experiments, opening their use to multiplexed single-molecule force spectroscopy assays. We propose that our technique will also be useful in other applications that involve the tracking of microscopic objects in three dimensions.

SIGNIFICANCETracking objects in 3D is a common task in biology, but typically requires the acquisition of image stacks, which is limited by speed, the depth of field of microscope objectives and by the presence of other objects that obscure the illumination. Here we develop HoloMiP (Holographic Microscopy with Parabolic masking), which uses digital holography to reconstruct the three-dimensional images from a single plane allowing tracking of light-scattering objects in 3D. HoloMiP outperforms existing methods in precision, speed, simplicity and tolerance to crowding. We show that it is particularly suitable for fast, multiplexed magnetic tweezer experiments, opening new avenues to high-throughput force spectroscopy.
]]></description>
<dc:creator>Tolar, P.</dc:creator>
<dc:creator>Flewellen, J.</dc:creator>
<dc:creator>Minoughan, S.</dc:creator>
<dc:creator>Llorente Garcia, I.</dc:creator>
<dc:date>2022-05-04</dc:date>
<dc:identifier>doi:10.1101/2022.05.03.490423</dc:identifier>
<dc:title><![CDATA[Digital holography-based 3D particle localisation for single molecule tweezer techniques]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.05.06.490861v1?rss=1">
<title>
<![CDATA[
Nucleation and stability of branched versus linear Arp2/3-generated actin filaments 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.05.06.490861v1?rss=1"
</link>
<description><![CDATA[
Activation of the Arp2/3 complex by VCA-domain-bearing NPFs results in the formation of  daughter actin filaments branching off the sides of pre-existing  mother filaments. Alternatively, when stimulated by SPIN90, Arp2/3 directly nucleates  linear actin filaments. Uncovering the similarities and differences of these two activation mechanisms is fundamental to understanding the regulation and function of Arp2/3. Analysis of individual filaments reveals that the catalytic VCA domain of WASP, N-WASP and WASH, accelerate the Arp2/3-mediated nucleation of linear filaments by SPIN90, in addition to their known branch-promoting activity. Unexpectedly, these VCA domains also destabilize existing branches, as well as SPIN90-Arp2/3 at filament pointed ends. Furthermore, cortactin and GMF, which respectively stabilize and destabilize Arp2/3 at branch junctions, have a similar impact on SPIN90-activated Arp2/3. However, unlike branch junctions, SPIN90-Arp2/3 at the pointed end of linear filaments is not destabilized by piconewton forces, and does not become less stable with time. It thus appears that linear and branched Arp2/3-generated filaments respond similarly to regulatory proteins, albeit with quantitative differences, and that they differ greatly in their responses to aging and to mechanical stress. These results indicate that SPIN90- and VCA-activated Arp2/3 complexes adopt similar yet non-identical conformations, and that their turnover in cells may be regulated differently.
]]></description>
<dc:creator>Cao, L.</dc:creator>
<dc:creator>Ghasemi, F.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:creator>Jegou, A.</dc:creator>
<dc:creator>Romet-Lemonne, G.</dc:creator>
<dc:date>2022-05-06</dc:date>
<dc:identifier>doi:10.1101/2022.05.06.490861</dc:identifier>
<dc:title><![CDATA[Nucleation and stability of branched versus linear Arp2/3-generated actin filaments]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.05.06.490863v1?rss=1">
<title>
<![CDATA[
Fluorescence fluctuation based super resolution microscopy, basic concepts for an easy start. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.05.06.490863v1?rss=1"
</link>
<description><![CDATA[
Due to the wave nature of light, optical microscopy has a lower-bound lateral resolution limit of about half of the wavelength of the detected light, i.e., within the range of 200 to 300 nm. The Fluorescence Fluctuation based Super Resolution Microscopy (FF-SRM) encompases a collection of image analysis techniques which rely on the statistical processing of temporal variations of fluorescence to reduce the uncertainty about the fluorophore positions within a sample, hence, bringing spatial resolution down to several tens of nm. The FF-SRM is known to be suitable for live-cell imaging due to its compatibility with most fluorescent probes and lower instrumental and experimental requirements, which are mostly camera-based epifluorescence instruments. Each FF-SRM approach has strengths and weaknesses, which depend directly on the underlying statistical principles through which enhanced spatial resolution is achieved. In this review, the basic concepts and principles behind a range of FF-SRM methods published to date are revisited. Their operational parameters are explained and guidance for its selection is provided.
]]></description>
<dc:creator>Alva, A.</dc:creator>
<dc:creator>Brito-Alarcon, E.</dc:creator>
<dc:creator>Linares, A.</dc:creator>
<dc:creator>Torres-Garcia, E.</dc:creator>
<dc:creator>Hernandez, H. O.</dc:creator>
<dc:creator>Pinto-Camara, R.</dc:creator>
<dc:creator>Martinez, D.</dc:creator>
<dc:creator>Hernandez-Herrera, P.</dc:creator>
<dc:creator>D'Antuono, R.</dc:creator>
<dc:creator>Wood, C.</dc:creator>
<dc:creator>Guerrero, A.</dc:creator>
<dc:date>2022-05-06</dc:date>
<dc:identifier>doi:10.1101/2022.05.06.490863</dc:identifier>
<dc:title><![CDATA[Fluorescence fluctuation based super resolution microscopy, basic concepts for an easy start.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.05.12.491749v1?rss=1">
<title>
<![CDATA[
Profile of Mycobacterium tuberculosis-specific CD4 T cells at the site of disease and blood in pericardial tuberculosis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.05.12.491749v1?rss=1"
</link>
<description><![CDATA[
Our understanding of the immune response at the site of disease in extra-pulmonary tuberculosis (EPTB) is still limited. In this study, using flow cytometry, we defined the pericardial fluid (PCF) cellular composition and compared the phenotypic and functional profile of Mycobacterium tuberculosis (Mtb)-specific T cells between PCF and whole blood in 16 patients with pericardial TB (PCTB). We found that lymphocytes were the predominant cell type in PCF in PCTB, with a preferential influx of CD4 T cells. The frequencies of TNF- producing myeloid cells and Mtb-specific T cells were significantly higher in PCF compared to blood. Mtb-specific CD4 T cells in PCF exhibited a distinct phenotype compared to those in blood, with greater GrB expression and lower CD27 and KLRG1 expression. We observed no difference in the production IFN{gamma}, TNF or IL-2 by Mtb-specific CD4 T cells between the two compartments, but MIP-1{beta} production was lower in the PCF T cells. Bacterial loads in the PCF did not relate to the phenotype or function of Mtb-specific CD4 T cells. Upon anti-tubercular treatment completion, HLA-DR, Ki-67 and GrB expression was significantly decreased, and relative IL-2 production was increased in peripheral Mtb-specific CD4 T cells. Overall, using a novel and rapid experimental approach to measure T cell response ex vivo at site of disease, these results provide novel insight into molecular mechanisms and immunopathology at site of TB infection of the pericardium.
]]></description>
<dc:creator>Du Bruyn, E.</dc:creator>
<dc:creator>Ruzive, S.</dc:creator>
<dc:creator>Howlett, P. J.</dc:creator>
<dc:creator>Jacobs, A. J.</dc:creator>
<dc:creator>Lindestam Arlehamn, C. S.</dc:creator>
<dc:creator>Sette, A.</dc:creator>
<dc:creator>Sher, A.</dc:creator>
<dc:creator>Mayer-Barber, K. D.</dc:creator>
<dc:creator>Barber, D.</dc:creator>
<dc:creator>Mayosi, B.</dc:creator>
<dc:creator>Ntsekhe, M.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Riou, C.</dc:creator>
<dc:date>2022-05-13</dc:date>
<dc:identifier>doi:10.1101/2022.05.12.491749</dc:identifier>
<dc:title><![CDATA[Profile of Mycobacterium tuberculosis-specific CD4 T cells at the site of disease and blood in pericardial tuberculosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.05.16.491352v1?rss=1">
<title>
<![CDATA[
From recency to the central tendency bias in working memory: a unifying attractor network model 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.05.16.491352v1?rss=1"
</link>
<description><![CDATA[
The central tendency bias, or contraction bias, is a phenomenon where the judgment of the magnitude of items held in working memory appears to be biased towards the average of past observations. It is assumed to be an optimal strategy by the brain, and commonly thought of as an expression of the brains ability to learn the statistical structure of sensory input. On the other hand, recency biases such as serial dependence are also commonly observed, and are thought to reflect the content of working memory. Recent results from an auditory delayed comparison task in rats, suggest that both biases may be more related than previously thought: when the posterior parietal cortex (PPC) was silenced, both short-term and contraction biases were reduced. By proposing a model of the circuit that may be involved in generating the behavior, we show that a volatile working memory content susceptible to shifting to the past sensory experience - producing short-term sensory history biases - naturally leads to contraction bias. The errors, occurring at the level of individual trials, are sampled from the full distribution of the stimuli, and are not due to a gradual shift of the memory towards the sensory distributions mean. Our results are consistent with a broad set of behavioral findings and provide predictions of performance across different stimulus distributions and timings, delay intervals, as well as neuronal dynamics in putative working memory areas. Finally, we validate our model by performing a set of human psychophysics experiments of an auditory parametric working memory task.
]]></description>
<dc:creator>Boboeva, V.</dc:creator>
<dc:creator>Pezzotta, A.</dc:creator>
<dc:creator>Akrami, A.</dc:creator>
<dc:creator>Clopath, C.</dc:creator>
<dc:date>2022-05-16</dc:date>
<dc:identifier>doi:10.1101/2022.05.16.491352</dc:identifier>
<dc:title><![CDATA[From recency to the central tendency bias in working memory: a unifying attractor network model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.05.16.492129v1?rss=1">
<title>
<![CDATA[
Cachd1 is a novel Frizzled- and LRP6-interacting protein required for neurons to acquire left-right asymmetric character 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.05.16.492129v1?rss=1"
</link>
<description><![CDATA[
Neurons on left and right sides of the nervous system frequently show asymmetric properties but how these differences arise is poorly understood. Through a forward genetic screen in zebrafish, we find that loss of function of the transmembrane protein Cachd1 results in right-sided habenula neurons adopting left-sided character. Cachd1 is expressed in habenula neuron progenitors, functions symmetrically downstream of asymmetric environmental signals that determine laterality and influences timing of the normally left-right asymmetric patterns of neurogenesis. Unbiased screening for Cachd1 partners identified the Wnt co-receptor Frizzled7 and further biochemical and structural analysis revealed Cachd1 can bind simultaneously to Fzd proteins and Lrp6, bridging between these two Wnt co-receptors. Consistent with these structural studies, lrp6 mutant zebrafish show symmetric habenulae with left-sided character and epistasis experiments with other Wnt pathway genes support an in vivo role for Cachd1 in modulating Wnt pathway activity in the brain. Together, these studies identify Cachd1 as a conserved novel Wnt-receptor interacting protein with roles in regulating neurogenesis and neuronal identity.
]]></description>
<dc:creator>Powell, G. T.</dc:creator>
<dc:creator>Zhao, Y.</dc:creator>
<dc:creator>Faro, A.</dc:creator>
<dc:creator>Stickney, H. L.</dc:creator>
<dc:creator>Novellasdemunt, L.</dc:creator>
<dc:creator>Henriques, P.</dc:creator>
<dc:creator>Gestri, G.</dc:creator>
<dc:creator>Redhouse White, E.</dc:creator>
<dc:creator>Ren, J.</dc:creator>
<dc:creator>Lu, W.</dc:creator>
<dc:creator>Young, R. M.</dc:creator>
<dc:creator>Hawkins, T. A.</dc:creator>
<dc:creator>Cavodeassi, F.</dc:creator>
<dc:creator>Schwarz, Q.</dc:creator>
<dc:creator>Dreosti, E.</dc:creator>
<dc:creator>Raible, D. W.</dc:creator>
<dc:creator>Li, V. S. W.</dc:creator>
<dc:creator>Wright, G. J.</dc:creator>
<dc:creator>Jones, E. Y.</dc:creator>
<dc:creator>Wilson, S. W.</dc:creator>
<dc:date>2022-05-17</dc:date>
<dc:identifier>doi:10.1101/2022.05.16.492129</dc:identifier>
<dc:title><![CDATA[Cachd1 is a novel Frizzled- and LRP6-interacting protein required for neurons to acquire left-right asymmetric character]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.05.17.492318v1?rss=1">
<title>
<![CDATA[
The Proteomic Landscape of Genome-Wide Genetic Perturbations 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.05.17.492318v1?rss=1"
</link>
<description><![CDATA[
Functional genomic strategies help to address the genotype phenotype problem by annotating gene function and regulatory networks. Here, we demonstrate that combining functional genomics with proteomics uncovers general principles of protein expression, and provides new avenues to annotate protein function. We recorded precise proteomes for all non-essential gene knock-outs in Saccharomyces cerevisiae. We find that protein abundance is driven by a complex interplay of i) general biological properties, including translation rate, turnover, and copy number variations, and ii) their genetic, metabolic and physical interactions, including membership in protein complexes. We further show that combining genetic perturbation with proteomics provides complementary dimensions of functional annotation: proteomic profiling, reverse proteomic profiling, profile similarity and protein covariation analysis. Thus, our study generates a resource in which nine million protein quantities are linked to 79% of the yeast coding genome, and shows that functional proteomics reveals principles that govern protein expression.

Highlights- Nine million protein quantities recorded in ~4,600 non-essential gene deletions in S. cerevisiae reveal principles of how the proteome responds to genetic perturbation
- Genome-scale protein expression is determined by both functional relationships between proteins, as well as common biological responses
- Broad protein expression profiles in slow-growing strains can be explained by chromosomal aneuploidies
- Protein half-life and ribosome occupancy are predictable from protein abundance changes across knock-outs
- Functional proteomics annotates missing gene function in four complementary dimensions
]]></description>
<dc:creator>Messner, C. B.</dc:creator>
<dc:creator>Demichev, V.</dc:creator>
<dc:creator>Muenzner, J.</dc:creator>
<dc:creator>Aulakh, S.</dc:creator>
<dc:creator>Röhl, A.</dc:creator>
<dc:creator>Herrera-DomInguez, L.</dc:creator>
<dc:creator>Egger, A.-S.</dc:creator>
<dc:creator>Kamrad, S.</dc:creator>
<dc:creator>Lemke, O.</dc:creator>
<dc:creator>Calvani, E.</dc:creator>
<dc:creator>Mülleder, M.</dc:creator>
<dc:creator>Lilley, K. S.</dc:creator>
<dc:creator>Kustatscher, G.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:date>2022-05-18</dc:date>
<dc:identifier>doi:10.1101/2022.05.17.492318</dc:identifier>
<dc:title><![CDATA[The Proteomic Landscape of Genome-Wide Genetic Perturbations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.05.19.492673v1?rss=1">
<title>
<![CDATA[
mNeonGreen-tagged fusion proteins and nanobodies reveal localization of tropomyosin to patches, cables, and contractile actomyosin rings in live yeast cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.05.19.492673v1?rss=1"
</link>
<description><![CDATA[
Tropomyosins are structurally conserved -helical coiled-coil dimeric proteins that bind along the length of filamentous actin (F-actin) in fungi and animals. Tropomyosins play essential roles in the stability of actin filaments in non-muscle cells and are essential for calcium regulation of myosin II contractility in the muscle. Despite the crucial role of tropomyosin in actin cytoskeletal regulation, in vivo investigations of tropomyosin are limited, mainly due to the suboptimal live cell imaging tools currently available in many organisms. Here, we report mNeon-Green (mNG) tagged tropomyosin, with native promoter and linker length configuration, that clearly reports tropomyosin localization and dynamics in Schizosaccharomyces pombe (Cdc8), Schizosaccharomyces japonicus (Cdc8), and Saccharomyces cerevisiae (Tpm1 and Tpm2), in vivo and in isolated S. pombe cell division apparatuses. We extended this approach to also visualize the mammalian TPM2 isoform. Finally, we generated a camelid-nanobody against S. pombe Cdc8, which mimics the localization of mNG-Cdc8 in vivo without significantly influencing cell growth and dynamics of actin cytoskeleton. Using these tools, we report the presence of tropomyosin in previously unappreciated patch-like structures in fission and budding yeasts, show flow of tropomyosin (F-actin) cables to the cytokinetic actomyosin ring, and identify rearrangements of the actin cytoskeleton during mating. These powerful tools and strategies will aid better analyses of tropomyosin and actin cables in vivo.
]]></description>
<dc:creator>Hatano, T.</dc:creator>
<dc:creator>Lim, T. C.</dc:creator>
<dc:creator>Billault-Chaumartin, I.</dc:creator>
<dc:creator>Dhar, A.</dc:creator>
<dc:creator>Gu, Y.</dc:creator>
<dc:creator>Massam-Wu, T.</dc:creator>
<dc:creator>Adhishesha, S.</dc:creator>
<dc:creator>Springall, L.</dc:creator>
<dc:creator>Scott, W.</dc:creator>
<dc:creator>Sivashanmugam, L.</dc:creator>
<dc:creator>Mishima, M.</dc:creator>
<dc:creator>Martin, S. G.</dc:creator>
<dc:creator>Oliferenko, S.</dc:creator>
<dc:creator>Palani, S.</dc:creator>
<dc:creator>BALASUBRAMANIAN, M. K.</dc:creator>
<dc:date>2022-05-20</dc:date>
<dc:identifier>doi:10.1101/2022.05.19.492673</dc:identifier>
<dc:title><![CDATA[mNeonGreen-tagged fusion proteins and nanobodies reveal localization of tropomyosin to patches, cables, and contractile actomyosin rings in live yeast cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.05.24.493130v1?rss=1">
<title>
<![CDATA[
PKN2 deficiency leads both to prenatal congenital cardiomyopathy and defective angiotensin II stress responses. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.05.24.493130v1?rss=1"
</link>
<description><![CDATA[
BackgroundThe protein kinase PKN2 is required for embryonic development, and PKN2 knockout mice die as a result of failure in expansion of mesoderm tissues, cardiac development and neural tube closure. In the adult, cardiomyocyte PKN2 and PKN1 (in combination) are required for cardiac adaptation to pressure-overload. The role of PKN2 in contractile cardiomyocytes during development and its role in the adult heart remain to be fully established.

MethodsWe used mice with cardiomyocyte-directed knockout of PKN2 or global PKN2 haploinsufficiency. Cardiac function and dimensions were assessed with high resolution episcopic microscopy, MRI, micro-CT and echocardiography. Biochemical and histological changes were assessed.

ResultsCardiomyocyte-directed PKN2 knockout embryos displayed striking abnormalities in the compact myocardium, with frequent myocardial clefts and diverticula, ventricular septal defects and abnormal heart shape. The sub-Mendelian homozygous knockout survivors developed cardiac failure. RNASeq data showed upregulation of PKN2 in patients with dilated cardiomyopathy, suggesting an involvement in adult heart disease. Given the rarity of homozygous survivors with cardiomyocyte-specific deletion of PKN2, this was explored using mice with constitutive heterozygous PKN2 knockout. Cardiac hypertrophy resulting from hypertension induced by angiotensin II was reduced in haploinsufficient PKN2 mice relative to wild-type littermates, with suppression of cardiomyocyte hypertrophy and cardiac fibrosis.

ConclusionsCardiomyocyte PKN2 is essential for heart development and formation of compact myocardium, and is also required for cardiac hypertrophy in hypertension. Thus, PKN signalling may offer therapeutic options for managing congenital and adult heart diseases.
]]></description>
<dc:creator>Marshall, J. J.</dc:creator>
<dc:creator>Cull, J. J.</dc:creator>
<dc:creator>Alharbi, H. O.</dc:creator>
<dc:creator>Zaw Thin, M.</dc:creator>
<dc:creator>Cooper, S. T.</dc:creator>
<dc:creator>Barrington, C.</dc:creator>
<dc:creator>Vanyai, H.</dc:creator>
<dc:creator>Snoeks, T.</dc:creator>
<dc:creator>Siow, B.</dc:creator>
<dc:creator>Suaarez-Bonnet, A.</dc:creator>
<dc:creator>Herbert, E.</dc:creator>
<dc:creator>Stuckey, D. J.</dc:creator>
<dc:creator>Cameron, A.</dc:creator>
<dc:creator>Prin, F.</dc:creator>
<dc:creator>Cook, A. C.</dc:creator>
<dc:creator>Priestnall, S. L.</dc:creator>
<dc:creator>Chotani, S.</dc:creator>
<dc:creator>Rackham, O. J.</dc:creator>
<dc:creator>Meijles, D. N.</dc:creator>
<dc:creator>Mohun, T.</dc:creator>
<dc:creator>Clerk, A.</dc:creator>
<dc:creator>Parker, P.</dc:creator>
<dc:date>2022-05-24</dc:date>
<dc:identifier>doi:10.1101/2022.05.24.493130</dc:identifier>
<dc:title><![CDATA[PKN2 deficiency leads both to prenatal congenital cardiomyopathy and defective angiotensin II stress responses.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.05.28.493848v1?rss=1">
<title>
<![CDATA[
Characterization of Plasmodium falciparum myosin B in complex with the calmodulin-like domain of its light chain MLC-B 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.05.28.493848v1?rss=1"
</link>
<description><![CDATA[
Myosin B (MyoB) is a class 14 myosin expressed in all invasive stages of the malaria parasite, Plasmodium falciparum. It is not associated with the glideosome complex that drives motility and invasion of host cells. During red blood cell invasion, it remains at the apical tip of the merozoite but is no longer observed once invasion is completed. MyoB is not essential for parasite survival but, when it is knocked out, merozoites are delayed in the initial stages of red blood cell invasion, giving rise to a growth defect that correlates with reduced invasion success. Here, we have expressed and purified functional MyoB with the help of parasite-specific chaperones Hsp90 and Unc45, characterized its binding to actin and its known light chain MLC-B using biochemical and biophysical methods, and determined its low-resolution structure in solution using small-angle X-ray scattering. In addition to MLC-B, four other putative regulatory light chains were found to bind to the MyoB IQ2 motif in vitro. The purified recombinant MyoB adopted the overall shape of a myosin, exhibited actin-activated ATPase activity, and moved actin filaments in vitro. The ADP release rate was faster than the ATP turnover number, and thus, does not appear to be rate-limiting. This, together with the observed high affinity to actin and the specific localization of MyoB, may point towards a role in tethering and/or force sensing during early stages of invasion.
]]></description>
<dc:creator>Pires, I.</dc:creator>
<dc:creator>Hung, Y.-F.</dc:creator>
<dc:creator>Bergmann, U.</dc:creator>
<dc:creator>Molloy, J. E.</dc:creator>
<dc:creator>Kursula, I.</dc:creator>
<dc:date>2022-05-28</dc:date>
<dc:identifier>doi:10.1101/2022.05.28.493848</dc:identifier>
<dc:title><![CDATA[Characterization of Plasmodium falciparum myosin B in complex with the calmodulin-like domain of its light chain MLC-B]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.05.31.494266v1?rss=1">
<title>
<![CDATA[
A nuclear export signal in KHNYN required for its antiviral activity evolved as ZAP emerged in tetrapods 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.05.31.494266v1?rss=1"
</link>
<description><![CDATA[
The zinc finger antiviral protein (ZAP) inhibits viral replication by directly binding CpG dinucleotides in cytoplasmic viral RNA to inhibit protein synthesis and target the RNA for degradation. ZAP evolved in tetrapods and there are clear orthologs in reptiles, birds and mammals. When ZAP emerged, other proteins may have evolved to become cofactors for its antiviral activity. KHNYN is a putative endoribonuclease that is required for ZAP to restrict retroviruses. To determine its evolutionary path after ZAP emerged, we compared KHNYN orthologs in mammals and reptiles to those in fish, which do not encode ZAP. This identified residues in KHNYN that are highly conserved in species that encode ZAP, including several in the CUBAN domain. The CUBAN domain interacts with NEDD8 and Cullin-RING E3 ubiquitin ligases. Deletion of the CUBAN domain decreased KHNYN antiviral activity, increased protein expression and increased nuclear localization. However, mutation of residues required for the CUBAN domain-NEDD8 interaction increased KHNYN abundance but did not affect its antiviral activity or cytoplasmic localization, indicating that Cullin- mediated degradation may control its homeostasis and regulation of protein turnover is separatable from its antiviral activity. By contrast, the C-terminal residues in the CUBAN domain form a CRM1-dependent nuclear export signal (NES) that is required for its antiviral activity. Deletion or mutation of the NES increased KHNYN nuclear localization and decreased its interaction with ZAP. The final two positions of this NES are not present in fish KHNYN orthologs and we hypothesize their evolution allowed KHNYN to act as a ZAP cofactor.

IMPORTANCEThe interferon system is part of the innate immune response that inhibits viruses and other pathogens. This system emerged approximately 500 million years ago in early vertebrates. Since then, some genes have evolved to become antiviral interferon- stimulated genes (ISGs) while others evolved so their encoded protein could interact with proteins encoded by ISGs and contribute to their activity. However, this remains poorly characterized. ZAP is an ISG that arose during tetrapod evolution and inhibits viral replication. Because KHNYN interacts with ZAP and is required for its antiviral activity against retroviruses, we conducted an evolutionary analysis to determine how specific amino acids in KHNYN evolved after ZAP emerged. This identified a nuclear export signal that evolved in tetrapods and is required for KHNYN to traffic in the cell to interact with ZAP. Overall, specific residues in KHNYN evolved to allow it to act as a cofactor for ZAP antiviral activity.
]]></description>
<dc:creator>Lista, M. J.</dc:creator>
<dc:creator>Ficarelli, M.</dc:creator>
<dc:creator>Wilson, H.</dc:creator>
<dc:creator>Kmiec, D.</dc:creator>
<dc:creator>Youle, R.</dc:creator>
<dc:creator>Wanford, J.</dc:creator>
<dc:creator>Winstone, H.</dc:creator>
<dc:creator>Odendall, C.</dc:creator>
<dc:creator>Taylor, I. A.</dc:creator>
<dc:creator>Neil, S. J.</dc:creator>
<dc:creator>Swanson, C. M.</dc:creator>
<dc:date>2022-06-01</dc:date>
<dc:identifier>doi:10.1101/2022.05.31.494266</dc:identifier>
<dc:title><![CDATA[A nuclear export signal in KHNYN required for its antiviral activity evolved as ZAP emerged in tetrapods]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.02.491151v1?rss=1">
<title>
<![CDATA[
Crosshair, semi-automated targeting for electron microscopy with a motorised ultramicrotome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.02.491151v1?rss=1"
</link>
<description><![CDATA[
Volume electron microscopy (EM) is a time consuming process - often requiring weeks or months of continuous acquisition for large samples. In order to compare the ultrastructure of a number of individuals or conditions, acquisition times must therefore be reduced. For resin-embedded samples, one solution is to selectively target smaller regions of interest by trimming with an ultramicrotome. This is a difficult and labour-intensive process, requiring manual positioning of the diamond knife and sample, and much time and training to master. Here, we have developed a semi-automated workflow for targeting with a modified ultramicrotome. We adapted two recent commercial systems to add motors for each rotational axis (and also each translational axis for one system), allowing precise and automated movement. We also developed a user-friendly software to convert X-ray images of resin-embedded samples into angles and cutting depths for the ultramicrotome. This is provided as an open-source Fiji plugin called Crosshair. This workflow is demonstrated by targeting regions of interest in a series of Platynereis dumerilii samples.
]]></description>
<dc:creator>Meechan, K.</dc:creator>
<dc:creator>Guan, W.</dc:creator>
<dc:creator>Riedinger, A.</dc:creator>
<dc:creator>Stankova, V.</dc:creator>
<dc:creator>Yoshimura, A.</dc:creator>
<dc:creator>Pipitone, R.</dc:creator>
<dc:creator>Milberger, A.</dc:creator>
<dc:creator>Schaar, H.</dc:creator>
<dc:creator>Romero-Brey, I.</dc:creator>
<dc:creator>Templin, R.</dc:creator>
<dc:creator>Peddie, C. J.</dc:creator>
<dc:creator>Schieber, N. L.</dc:creator>
<dc:creator>Jones, M. L.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:creator>Schwab, Y.</dc:creator>
<dc:date>2022-06-02</dc:date>
<dc:identifier>doi:10.1101/2022.06.02.491151</dc:identifier>
<dc:title><![CDATA[Crosshair, semi-automated targeting for electron microscopy with a motorised ultramicrotome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.01.494365v1?rss=1">
<title>
<![CDATA[
Diacylglycerol consumption at the inner nuclear membrane fuels nuclear envelope expansion required for closed mitosis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.01.494365v1?rss=1"
</link>
<description><![CDATA[
Nuclear envelope (NE) expansion must be controlled to maintain nuclear shape and function. The nuclear membrane expands massively during  closed mitosis, enabling chromosome segregation within an intact NE. Phosphatidic acid (PA) and diacylglycerol (DG) can both serve as biosynthetic precursors for membrane lipid synthesis. How they are regulated in time and space and what are the implications of changes in their flux for mitotic fidelity is largely unknown. Using genetically encoded PA and DG probes, we show that DG is depleted from the inner nuclear membrane during mitosis in the fission yeast Schizosaccharomyces pombe, but PA does not accumulate, indicating that it is rerouted to membrane synthesis. We demonstrate that DG-to-PA conversion catalysed by the diacylglycerol kinase Dgk1 and direct glycerophospholipid synthesis from DG by diacylglycerol cholinephosphotransferase / ethanolaminephosphotransferase Ept1 reinforce NE expansion. We conclude that DG consumption through both de novo and the Kennedy pathways fuels a spike in glycerophospholipid biosynthesis, controlling NE expansion, and ultimately, mitotic fidelity.
]]></description>
<dc:creator>Foo, S.</dc:creator>
<dc:creator>Cazenave-Gassiot, A.</dc:creator>
<dc:creator>Wenk, M. R.</dc:creator>
<dc:creator>Oliferenko, S.</dc:creator>
<dc:date>2022-06-02</dc:date>
<dc:identifier>doi:10.1101/2022.06.01.494365</dc:identifier>
<dc:title><![CDATA[Diacylglycerol consumption at the inner nuclear membrane fuels nuclear envelope expansion required for closed mitosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.01.494393v1?rss=1">
<title>
<![CDATA[
OxoScan-MS: Oxonium ion scanning mass spectrometry facilitates plasma glycoproteomics in large scale 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.01.494393v1?rss=1"
</link>
<description><![CDATA[
Protein glycosylation is a complex and heterogeneous post-translational modification. Specifically, the human plasma proteome is rich in glycoproteins, and as protein glycosylation is frequently dysregulated in disease, glycoproteomics is considered an underexplored resource for biomarker discovery. Here, we present OxoScan-MS, a data-independent mass spectrometric acquisition technology and data analysis software that facilitates sensitive, fast, and cost-effective glycoproteome profiling of plasma and serum samples in large cohort studies. OxoScan-MS quantifies glycosylated peptide features by exploiting a scanning quadrupole to assign precursors to oxonium ions, glycopeptide-specific fragments. OxoScan-MS reaches a high level of sensitivity and selectivity in untargeted glycopeptide profiling, such that it can be efficiently used with fast microflow chromatography without a need for experimental enrichment of glycopeptides from neat plasma. We apply OxoScan-MS to profile the plasma glycoproteomic in an inpatient cohort hospitalised due to severe COVID-19, and obtain precise quantities for 1,002 glycopeptide features. We reveal that severe COVID-19 induces differential glycosylation in disease-relevant plasma glycoproteins, including IgA, fibrinogen and alpha-1-antitrypsin. Thus, with OxoScan-MS we present a strategy for quantitatively mapping glycoproteomes that scales to hundreds and thousands of samples, and report glycoproteomic changes in severe COVID-19.
]]></description>
<dc:creator>White, M. E. H.</dc:creator>
<dc:creator>Jones, D. M.</dc:creator>
<dc:creator>de Folter, J.</dc:creator>
<dc:creator>Aulakh, S. K.</dc:creator>
<dc:creator>Flynn, H. R.</dc:creator>
<dc:creator>Krüger, L.</dc:creator>
<dc:creator>Demichev, V.</dc:creator>
<dc:creator>Tober-Lau, P.</dc:creator>
<dc:creator>Kurth, F.</dc:creator>
<dc:creator>Mülleder, M.</dc:creator>
<dc:creator>Blanchard, V.</dc:creator>
<dc:creator>Messner, C. B.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:date>2022-06-02</dc:date>
<dc:identifier>doi:10.1101/2022.06.01.494393</dc:identifier>
<dc:title><![CDATA[OxoScan-MS: Oxonium ion scanning mass spectrometry facilitates plasma glycoproteomics in large scale]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.03.494621v1?rss=1">
<title>
<![CDATA[
Reconstituting human somitogenesis in vitro 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.03.494621v1?rss=1"
</link>
<description><![CDATA[
The segmented body plan of vertebrates is established during somitogenesis, a well-studied process in model organisms, but remains largely elusive in humans due to ethical and technical limitations. Despite recent advances with pluripotent stem cell (PSC)-based approaches1-5, a system that robustly recapitulates human somitogenesis in both space and time remains missing. Here, we introduce a PSC-derived mesoderm-based 3D model of human segmentation and somitogenesis, which we termed Axioloids, that captures accurately the oscillatory dynamics of the segmentation clock as well as the morphological and molecular characteristics of segmentation and sequential somite formation in vitro. Axioloids show proper rostrocaudal patterning of forming segments and robust anterior-posterior FGF/WNT signaling gradients and Retinoic Acid (RA) signaling components. We identify an unexpected critical role of RA signaling in the stabilization of forming segments, indicating distinct, but also synergistic effects of RA and extracellular matrix (ECM) on the formation and epithelialization of somites. Importantly, comparative analysis demonstrates striking similarities of Axioloids to the human embryo, further validated by the presence of the HOX code in Axioloids. Lastly, we demonstrate the utility of our Axioloid system to study the pathogenesis of human congenital spine diseases, by using patient-like iPSC cells with mutations in HES7 and MESP2, which revealed disease-associated phenotypes including loss of epithelial somite formation and abnormal rostrocaudal patterning. These results suggest that Axioloids represent a promising novel platform to study axial development and disease in humans.
]]></description>
<dc:creator>Yamanaka, Y.</dc:creator>
<dc:creator>Yoshioka-Kobayashi, K.</dc:creator>
<dc:creator>Hamidi, S.</dc:creator>
<dc:creator>Munira, S.</dc:creator>
<dc:creator>Sunadome, K.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Kurokawa, Y.</dc:creator>
<dc:creator>Mieda, A.</dc:creator>
<dc:creator>Thompson, J. L.</dc:creator>
<dc:creator>Kerwin, J.</dc:creator>
<dc:creator>Lisgo, S.</dc:creator>
<dc:creator>Yamamoto, T.</dc:creator>
<dc:creator>Moris, N.</dc:creator>
<dc:creator>Martinez-Arias, A.</dc:creator>
<dc:creator>Tsujimura, T.</dc:creator>
<dc:creator>Alev, C.</dc:creator>
<dc:date>2022-06-03</dc:date>
<dc:identifier>doi:10.1101/2022.06.03.494621</dc:identifier>
<dc:title><![CDATA[Reconstituting human somitogenesis in vitro]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.03.494608v1?rss=1">
<title>
<![CDATA[
Taxonium: a web-based tool for exploring large phylogenetic trees 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.03.494608v1?rss=1"
</link>
<description><![CDATA[
The COVID-19 pandemic has resulted in a step change in the scale of sequencing data, with more genomes of SARS-CoV-2 having been sequenced than any other organism on earth. These sequences reveal key insights when represented as a phylogenetic tree, which captures the evolutionary history of the virus, and allows the identification of transmission events and the emergence of new variants. However, existing web-based tools for exploring phylogenies do not scale to the size of datasets now available for SARS-CoV-2. We have developed Taxonium, a new tool that uses WebGL to allow the exploration of trees with tens of millions of nodes in the browser for the first time. Taxonium links each node to associated metadata and supports mutation-annotated trees, which are able to capture all known genetic variation in a dataset. It can either be run entirely locally in the browser, from a server-based backend, or as a desktop application. We describe insights that analysing a tree of five million sequences can provide into SARS-CoV-2 evolution, and provide a tool at cov2tree.org for exploring a public tree of more than five million SARS-CoV-2 sequences. Taxonium can be applied to any tree, and is available at taxonium.org, with source code at github.com/theosanderson/taxonium.
]]></description>
<dc:creator>Sanderson, T.</dc:creator>
<dc:date>2022-06-03</dc:date>
<dc:identifier>doi:10.1101/2022.06.03.494608</dc:identifier>
<dc:title><![CDATA[Taxonium: a web-based tool for exploring large phylogenetic trees]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.03.494659v1?rss=1">
<title>
<![CDATA[
Bumetanide increases microglia-interneuron contact following traumatic brain injury 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.03.494659v1?rss=1"
</link>
<description><![CDATA[
ObjectiveThe Na-K-Cl cotransporter (NKCC1) inhibitor bumetanide has prominent positive effects on the pathophysiology of many neurological disorders. Here we studied whether bumetanide could influence post-traumatic cognitive decline and inflammatory processes by regulating astrocyte and microglia activation.

MethodControlled cortical impacted (CCI) animals were treated with bumetanide during the first post-CCI week. Immunochemistry, flow cytometry, immunoassay, and in vivo imaging were used to study astrocytic and microglial morphology and phenotype as well as adult neurogenesis. Telemetric electroencephalograms and cognitive behavioral test were performed at one-month post CCI.

ResultsBumetanide prevented CCI-induced decrease in hippocampal neurogenesis and parvalbumin positive interneuron loss. Deletion of NKCC1 in astrocytes neither rescued interneurons nor promote neurogenesis. Interestingly, bumetanide had a strong effect on microglial activation by inducing polarization towards the M1-like phenotype 3 days post-CCI and the M2-like phenotype 7 days post-CCI. Bumetanide increased microglial Brain-derived neurotrophic factor (BDNF) expression and interaction with parvalbumin interneurons. The early treatment with bumetanide resulted in improvements in working and episodic memory, one-month post-CCI, as well as the normalization of theta band oscillations.

InterpretationHere, we disclose a novel mechanism for the neuroprotective action of bumetanide mediated by an acceleration of microglial activation dynamics that leads to an increase of parvalbumin interneuron survival following CCI, possibly resulting from increased microglial BDNF expression and contact with interneurons. Salvage of interneurons may normalize ambient gamma-aminobutyric acid (GABA) resulting in the preservation of adult neurogenesis processes as well as contributing to bumetanide-mediated improvement of cognitive performance.
]]></description>
<dc:creator>Tessier, M.</dc:creator>
<dc:creator>Garcia, M. S.</dc:creator>
<dc:creator>Goubert, E.</dc:creator>
<dc:creator>Tian, L.</dc:creator>
<dc:creator>Molinari, F.</dc:creator>
<dc:creator>Blasco, E.</dc:creator>
<dc:creator>Laurin, J.</dc:creator>
<dc:creator>Guillemot, F.</dc:creator>
<dc:creator>Hubner, C.</dc:creator>
<dc:creator>Pellegrino, c. A.</dc:creator>
<dc:creator>Rivera, C.</dc:creator>
<dc:date>2022-06-05</dc:date>
<dc:identifier>doi:10.1101/2022.06.03.494659</dc:identifier>
<dc:title><![CDATA[Bumetanide increases microglia-interneuron contact following traumatic brain injury]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.06.494792v1?rss=1">
<title>
<![CDATA[
Developmental cell fate choice employs two distinct cis regulatory strategies 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.06.494792v1?rss=1"
</link>
<description><![CDATA[
In many developing tissues the patterns of gene expression that assign cell fate are organised by secreted signals functioning in a graded manner over multiple cell diameters. Cis Regulatory Elements (CREs) interpret these graded inputs to control gene expression. How this is accomplished remains poorly understood. In the neural tube, a gradient of the morphogen Sonic hedgehog allocates neural progenitor identity. Here, we uncover two distinct ways in which CREs translate graded Shh signaling into differential gene expression. In the majority of ventral neural progenitors a common set of CREs are used to control gene activity. These CREs integrate cell type specific inputs to control gene expression. By contrast, the most ventral progenitors use a unique set of CREs. These are established by the pioneer factor FOXA2, paralleling the role of FOXA2 in endoderm. Moreover, FOXA2 binds a subset of the same sites in neural and endoderm cells. Together the data identify distinct cis regulatory strategies for the interpretation of morphogen signaling and raise the possibility of an evolutionarily conserved role for FOXA2-mediated regulatory strategy across tissues.
]]></description>
<dc:creator>Delas, M. J.</dc:creator>
<dc:creator>Kalaitzis, C. M.</dc:creator>
<dc:creator>Fawzi, T.</dc:creator>
<dc:creator>Demuth, M.</dc:creator>
<dc:creator>Zhang, I.</dc:creator>
<dc:creator>Stuart, H. T.</dc:creator>
<dc:creator>Costantini, E.</dc:creator>
<dc:creator>Ivanovitch, K.</dc:creator>
<dc:creator>Tanaka, E.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:date>2022-06-06</dc:date>
<dc:identifier>doi:10.1101/2022.06.06.494792</dc:identifier>
<dc:title><![CDATA[Developmental cell fate choice employs two distinct cis regulatory strategies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.07.494932v1?rss=1">
<title>
<![CDATA[
Structural conversion of α-synuclein at the mitochondria induces neuronal toxicity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.07.494932v1?rss=1"
</link>
<description><![CDATA[
Aggregation of -Synuclein (-Syn) drives Parkinsons disease, although the initial stages of self-assembly and structural conversion have not been captured inside neurons. We track the intracellular conformational states of -Syn utilizing a single-molecule FRET biosensor, and show that -Syn converts from its monomeric state to form two distinct oligomeric states in neurons in a concentration dependent, and sequence specific manner. 3D FRET-CLEM reveals the structural organization, and location of aggregation hotspots inside the cell. Notably multiple intracellular seeding events occur preferentially on membrane surfaces, especially mitochondrial membranes. The mitochondrial lipid, cardiolipin triggers rapid oligomerization of A53T -Syn, and cardiolipin is sequestered within aggregating lipid-protein complexes. Mitochondrial aggregates impair complex I activity and increase mitochondrial ROS generation, which accelerates the oligomerization of A53T -Syn, and ultimately causes permeabilization of mitochondrial membranes, and cell death. Patient iPSC derived neurons harboring A53T mutations exhibit accelerated oligomerization that is dependent on mitochondrial ROS, early mitochondrial permeabilization and neuronal death. Our study highlights a mechanism of de novo oligomerization at the mitochondria and its induction of neuronal toxicity.
]]></description>
<dc:creator>Choi, M. L.</dc:creator>
<dc:creator>Chappard, A.</dc:creator>
<dc:creator>Singh, B. P.</dc:creator>
<dc:creator>Maclachlan, C.</dc:creator>
<dc:creator>Rodrigues, M.</dc:creator>
<dc:creator>Fedotova, E.</dc:creator>
<dc:creator>Berezhnov, A.</dc:creator>
<dc:creator>De, S.</dc:creator>
<dc:creator>Peddie, C.</dc:creator>
<dc:creator>Athauda, D.</dc:creator>
<dc:creator>Virdi, G. S.</dc:creator>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>Wernick, A.</dc:creator>
<dc:creator>Zanjani, Z.</dc:creator>
<dc:creator>Angelova, P. R.</dc:creator>
<dc:creator>Esteras, N.</dc:creator>
<dc:creator>Andrey, A.</dc:creator>
<dc:creator>Morris, K.</dc:creator>
<dc:creator>Jeacock, K.</dc:creator>
<dc:creator>Tosatto, L.</dc:creator>
<dc:creator>Little, D.</dc:creator>
<dc:creator>Gissen, P.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:creator>Clarke, D. J.</dc:creator>
<dc:creator>Kunath, T.</dc:creator>
<dc:creator>Klenerman, D.</dc:creator>
<dc:creator>Abramov, A. Y.</dc:creator>
<dc:creator>Horrocks, M. H.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:date>2022-06-09</dc:date>
<dc:identifier>doi:10.1101/2022.06.07.494932</dc:identifier>
<dc:title><![CDATA[Structural conversion of α-synuclein at the mitochondria induces neuronal toxicity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.10.495627v1?rss=1">
<title>
<![CDATA[
Cis-membrane association of human ATG8 proteins N-terminus mediates autophagy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.10.495627v1?rss=1"
</link>
<description><![CDATA[
Autophagy is an essential catabolic pathway which sequesters and engulfs cytosolic substrates via autophagosomes, unique double-membraned structures. ATG8 proteins are ubiquitin-like proteins recruited to autophagosome membranes by lipidation at the C-terminus. ATG8s recruit substrates, such as p62, and play an important role in mediating autophagosome membrane expansion. However, the precise function of lipidated ATG8 in expansion remains obscure. Using a real-time in vitro lipidation assay, we revealed that the N-termini of lipidated human ATG8s (LC3B and GABARAP) are highly dynamic and interact with the membrane. Moreover, atomistic MD simulation and FRET assays indicate that N-termini of LC3B and GABARAP associate in cis on the membrane. The cis-membrane association of the N-terminus is critical to maintain membrane expansion and the size of autophagosomes in cells, consequently, mediating the efficient degradation of p62. Our study provides fundamental molecular insights into autophagosome membrane expansion, revealing the critical and unique function of lipidated ATG8.
]]></description>
<dc:creator>Tooze, S.</dc:creator>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Nishimura, T.</dc:creator>
<dc:creator>Gahlot, D.</dc:creator>
<dc:creator>Saito, C.</dc:creator>
<dc:creator>Davis, C.</dc:creator>
<dc:creator>Jefferies, H. B. J.</dc:creator>
<dc:creator>Schreiber, A.</dc:creator>
<dc:creator>Thukral, L.</dc:creator>
<dc:date>2022-06-10</dc:date>
<dc:identifier>doi:10.1101/2022.06.10.495627</dc:identifier>
<dc:title><![CDATA[Cis-membrane association of human ATG8 proteins N-terminus mediates autophagy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.08.494952v1?rss=1">
<title>
<![CDATA[
Quantitative analysis of rabies virus-based synaptic connectivity tracing 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.08.494952v1?rss=1"
</link>
<description><![CDATA[
Monosynaptically restricted rabies viruses have been used for more than a decade for synaptic connectivity tracing. However, the verisimilitude of quantitative conclusions drawn from these experiments is largely unknown. The primary reason is the simple metrics commonly used, which generally disregard the effect of starter cell numbers. Here we present an experimental dataset with a broad range of starter cell numbers and explore their relationship with the number of input cells across the brain using descriptive statistics and modelling. We show that starter cell numbers strongly affect input fraction and convergence index measures, making quantitative comparisons unreliable. Furthermore, we suggest a principled way to analyse rabies derived connectivity data by taking advantage of the starter vs input cell relationship that we describe and validate across independent datasets.
]]></description>
<dc:creator>Tran-Van-Minh, A.</dc:creator>
<dc:creator>Ye, Z.</dc:creator>
<dc:creator>Rancz, E.</dc:creator>
<dc:date>2022-06-10</dc:date>
<dc:identifier>doi:10.1101/2022.06.08.494952</dc:identifier>
<dc:title><![CDATA[Quantitative analysis of rabies virus-based synaptic connectivity tracing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.10.495597v1?rss=1">
<title>
<![CDATA[
Neutrophil degranulation, NETosis and platelet degranulation pathway genes are co-induced in whole blood up to six months before tuberculosis diagnosis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.10.495597v1?rss=1"
</link>
<description><![CDATA[
Mycobacterium tuberculosis (M.tb) causes tuberculosis (TB) and remains one of the leading causes of mortality due to an infectious pathogen. Host immune responses have been implicated in driving the progression from infection to severe lung disease. We analyzed longitudinal RNA sequencing (RNAseq) data from the whole blood of 74 TB progressors whose samples were grouped into four six-month intervals preceding diagnosis (the GC6-74 study). We additionally analyzed RNAseq data from an independent cohort of 90 TB patients with positron emission tomography-computed tomography (PET-CT) scan results which were used to categorize them into groups with high and low levels of lung damage (the Catalysis TB Biomarker study). These groups were compared to non-TB controls to obtain a complete whole blood transcriptional profile for individuals spanning from early stages of M.tb infection to TB diagnosis. The results revealed a steady increase in the number of genes that were differentially expressed in progressors at time points closer to diagnosis with 278 genes at 13-18 months, 742 at 7-12 months and 5,131 detected 1-6 months before diagnosis and 9,205 detected in TB patients. A total of 2,144 differentially expressed genes were detected when comparing TB patients with high and low levels of lung damage. There was a large overlap in the genes upregulated in progressors 1-6 months before diagnosis (86%) with those in TB patients. A comprehensive pathway analysis revealed a potent activation of neutrophil and platelet mediated defenses including neutrophil and platelet degranulation, and NET formation at both time points. These pathways were also enriched in TB patients with high levels of lung damage compared to those with low. These findings suggest that neutrophils and platelets play a critical role in TB pathogenesis, and provide details of the timing of specific effector mechanisms that may contribute to TB lung pathology.

Author summaryMycobacterium tuberculosis (M.tb) causes tuberculosis (TB) and remains one of the leading causes of mortality due to an infectious pathogen. Human immune responses must be balanced to inhibit disease progression while limiting self-damage, however in defense against M.tb host responses have been implicated in lung damage and in driving progression of M.tb infection to severe lung disease. The identification of immune responses that are activated during the development of TB could provide potential targets for interventions that might suppress disease progression, and possibly limit tissue damage. Here we identify a subset of genes that function in several biological processes that are strongly activated in the late stages of TB development and in TB patients. The proteins encoded by these genes are known to degrade lung tissue and contribute to severe lung disease.
]]></description>
<dc:creator>Meier, S.</dc:creator>
<dc:creator>Seddon, J. A.</dc:creator>
<dc:creator>Maasdorp, E.</dc:creator>
<dc:creator>Kleynhans, L.</dc:creator>
<dc:creator>du Plessis, N.</dc:creator>
<dc:creator>Loxton, A. G.</dc:creator>
<dc:creator>Malherbe, S. T.</dc:creator>
<dc:creator>Zak, D. E.</dc:creator>
<dc:creator>Thompson, E.</dc:creator>
<dc:creator>Duffy, F.</dc:creator>
<dc:creator>Kaufmann, S. H. E.</dc:creator>
<dc:creator>Ottenhoff, T. H. M.</dc:creator>
<dc:creator>Scriba, T. J.</dc:creator>
<dc:creator>Suliman, S.</dc:creator>
<dc:creator>Sutherland, J. S.</dc:creator>
<dc:creator>Winter, J.</dc:creator>
<dc:creator>Kuivaniemi, H.</dc:creator>
<dc:creator>Walzl, G.</dc:creator>
<dc:creator>Tromp, G.</dc:creator>
<dc:creator>GC6-74 Consortium,</dc:creator>
<dc:creator>Catalysis TB Biomarkers Consortium,</dc:creator>
<dc:date>2022-06-11</dc:date>
<dc:identifier>doi:10.1101/2022.06.10.495597</dc:identifier>
<dc:title><![CDATA[Neutrophil degranulation, NETosis and platelet degranulation pathway genes are co-induced in whole blood up to six months before tuberculosis diagnosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.09.495437v1?rss=1">
<title>
<![CDATA[
A low-cost device for cryoanesthesia of neonate rodents 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.09.495437v1?rss=1"
</link>
<description><![CDATA[
Studying the development of neural circuits in rodent models requires surgical access to the neonatal brain. Since commercially available stereotaxic and anesthetic equipment is designed for use in adults, reliable targeting of brain structures in such young animals can be challenging. Hypothermic cooling (cryoanesthesia) has been used as a preferred anesthesia approach in neonates. This commonly involves submerging neonates in ice, an approach that is poorly controllable. We have developed an affordable, simple to construct device - CryoPup - that allows for fast and robust cryoanesthesia of rodent pups. CryoPup consists of a microcontroller controlling a Peltier element and a heat exchanger. It is capable of both cooling and heating, thereby also functioning as a heating pad during recovery. Importantly, it has been designed for size compatibility with common stereotaxic frames. We validate CryoPup in neonatal mice, demonstrating that it allows for rapid, reliable and safe cryoanesthesia and subsequent recovery. This open-source device will facilitate future studies into the development of neural circuits in the postnatal brain.

Specifications table

O_TBL View this table:
org.highwire.dtl.DTLVardef@de1fdorg.highwire.dtl.DTLVardef@16fde69org.highwire.dtl.DTLVardef@172f48corg.highwire.dtl.DTLVardef@15454forg.highwire.dtl.DTLVardef@79676e_HPS_FORMAT_FIGEXP  M_TBL C_TBL
]]></description>
<dc:creator>Jamieson, B. B.</dc:creator>
<dc:creator>Konstantinou, G.</dc:creator>
<dc:creator>Cano-Ferrer, X.</dc:creator>
<dc:creator>de Launoit, E.</dc:creator>
<dc:creator>Renier, N.</dc:creator>
<dc:creator>Imbert, A.</dc:creator>
<dc:creator>Kohl, J.</dc:creator>
<dc:date>2022-06-12</dc:date>
<dc:identifier>doi:10.1101/2022.06.09.495437</dc:identifier>
<dc:title><![CDATA[A low-cost device for cryoanesthesia of neonate rodents]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.10.495115v1?rss=1">
<title>
<![CDATA[
β-catenin obstructs γδ T cell immunosurveillance in colon cancer through loss of BTNL expression 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.10.495115v1?rss=1"
</link>
<description><![CDATA[
WNT/{beta}-catenin signaling endows cancer cells with proliferative capacity and immune-evasive functions that impair anti-cancer immunosurveillance by conventional, cytoxtoic T cells. However, the impact of dysregulated WNT signalling on unconventional, tissue-resident T cells, specifically in colon cancer is unknown. Here, we show that cancer cells in Apc-mutant mouse models escape immunosurveillance from gut-resident intraepithelial lymphocytes (IELs) expressing {gamma}{delta} T cell receptors ({gamma}{delta}TCRs). Analysis of late-stage tumors from mice and humans revealed that {gamma}{delta}IELs are largely absent from the tumor microenvironment, and that butyrophilin-like (BTNL) molecules, which can critically regulate {gamma}{delta}IEL through direct {gamma}{delta}TCR-interactions, are also downregulated. We could attribute this to {beta}-catenin stabilization, which rapidly decreased expression of the transcription factors, HNF4A and HNF4G, that we found to bind promoter regions of Btnl genes, thereby driving their expression in normal gut epithelial cells. Indeed, inhibition of {beta}-catenin signaling restored Btnl1 gene expression and {gamma}{delta} T cell infiltration into tumors. These observations highlight an immune-evasion mechanism specific to WNT-driven colon cancer cells that disrupts {gamma}{delta}IEL immunosurveillance and furthers cancer progression.
]]></description>
<dc:creator>Suzuki, T.</dc:creator>
<dc:creator>Kilbey, A.</dc:creator>
<dc:creator>Ridgway, R. A.</dc:creator>
<dc:creator>Hayman, H.</dc:creator>
<dc:creator>Byrne, R.</dc:creator>
<dc:creator>Casa Rodriguez, N.</dc:creator>
<dc:creator>Georgakopoulou, A.</dc:creator>
<dc:creator>Chen, L.</dc:creator>
<dc:creator>Verzi, M.</dc:creator>
<dc:creator>Gay, D. M.</dc:creator>
<dc:creator>Vzquez, E. G.</dc:creator>
<dc:creator>Belnoue-Davis, H. L.</dc:creator>
<dc:creator>Gilroy, K.</dc:creator>
<dc:creator>Kostner, A. H.</dc:creator>
<dc:creator>Kersten, C.</dc:creator>
<dc:creator>Thuwajit, C.</dc:creator>
<dc:creator>Andersen, D.</dc:creator>
<dc:creator>Wiesheu, R.</dc:creator>
<dc:creator>Jandke, A.</dc:creator>
<dc:creator>Roberts, N.</dc:creator>
<dc:creator>Blyth, K.</dc:creator>
<dc:creator>Roseweir, A.</dc:creator>
<dc:creator>Leedham, S. J.</dc:creator>
<dc:creator>Dunne, P. D.</dc:creator>
<dc:creator>Edwards, J.</dc:creator>
<dc:creator>Hayday, A.</dc:creator>
<dc:creator>Sansom, O. J.</dc:creator>
<dc:creator>Coffelt, S. B.</dc:creator>
<dc:date>2022-06-12</dc:date>
<dc:identifier>doi:10.1101/2022.06.10.495115</dc:identifier>
<dc:title><![CDATA[β-catenin obstructs γδ T cell immunosurveillance in colon cancer through loss of BTNL expression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.13.495866v1?rss=1">
<title>
<![CDATA[
Inhibition of vaccinia virus L1 N-myristoylation by the host N-myristoyltransferase inhibitor IMP-1088 generates non-infectious virions defective in cell entry 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.13.495866v1?rss=1"
</link>
<description><![CDATA[
We have recently shown that the replication of rhinovirus, poliovirus and foot-and-mouth disease virus requires the co-translational N-myristoylation of viral proteins by human host cell N-myristoyltransferases (NMTs), and is inhibited by treatment with IMP-1088, an ultrapotent small molecule NMT inhibitor. Here, we reveal the role of N-myristoylation during vaccinia virus (VACV) infection in human host cells and demonstrate the anti-poxviral effects of IMP-1088. N-myristoylated proteins from VACV and the host were metabolically labelled with myristic acid alkyne during infection using quantitative chemical proteomics. We identified VACV proteins A16, G9 and L1 to be N-myristoylated. Treatment with NMT inhibitor IMP-1088 potently abrogated VACV infection, while VACV gene expression, DNA replication, morphogenesis and EV formation remained unaffected. Importantly, we observed that loss of N-myristoylation resulted in greatly reduced infectivity of assembled mature virus particles, characterized by significantly reduced host cell entry and a decline in membrane fusion activity of progeny virus. While the N-myristoylation of VACV entry proteins L1, A16 and G9 was inhibited by IMP-1088, mutational and genetic studies demonstrated that the N-myristoylation of L1 was the most critical for VACV entry. Given the significant genetic identity between VACV, monkeypox virus and variola virus L1 homologs, our data provides a basis for further investigating the role of N-myristoylation in poxviral infections as well as the potential of selective NMT inhibitors like IMP-1088 as broad-spectrum poxvirus inhibitors.
]]></description>
<dc:creator>Priyamvada, L.</dc:creator>
<dc:creator>Kallemeijn, W. W.</dc:creator>
<dc:creator>Faronato, M.</dc:creator>
<dc:creator>Wilkins, K.</dc:creator>
<dc:creator>Goldsmith, C. S.</dc:creator>
<dc:creator>Cotter, C. A.</dc:creator>
<dc:creator>Ojeda, S.</dc:creator>
<dc:creator>Solari, R.</dc:creator>
<dc:creator>Moss, B.</dc:creator>
<dc:creator>Tate, E. W.</dc:creator>
<dc:creator>Satheshkumar, P. S.</dc:creator>
<dc:date>2022-06-13</dc:date>
<dc:identifier>doi:10.1101/2022.06.13.495866</dc:identifier>
<dc:title><![CDATA[Inhibition of vaccinia virus L1 N-myristoylation by the host N-myristoyltransferase inhibitor IMP-1088 generates non-infectious virions defective in cell entry]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.10.17.464398v1?rss=1">
<title>
<![CDATA[
Nanoscopic resolution within a single imaging frame 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.10.17.464398v1?rss=1"
</link>
<description><![CDATA[
Mean-Shift Super Resolution (MSSR) is a principle based on the Mean Shift theory that extends spatial resolution in fluorescence images, beyond the diffraction limit. MSSR works on low- and high-density fluorophore images, is not limited by the architecture of the detector (EM-CCD, sCMOS, or photomultiplier-based laser scanning systems) and is applicable to single images as well as temporal series. The theoretical limit of spatial resolution, based on optimized real-world imaging conditions and analysis of temporal image series, has been measured to be 40 nm. Furthermore, MSSR has denoising capabilities that outperform other analytical super resolution image approaches. Altogether, MSSR is a powerful, flexible, and generic tool for multidimensional and live cell imaging applications.
]]></description>
<dc:creator>Torres, E.</dc:creator>
<dc:creator>Pinto, R.</dc:creator>
<dc:creator>Linares, A.</dc:creator>
<dc:creator>Martinez, D.</dc:creator>
<dc:creator>Abonza, V.</dc:creator>
<dc:creator>Brito-Alarcon, E.</dc:creator>
<dc:creator>Calcines-Cruz, C.</dc:creator>
<dc:creator>Galindo, G.</dc:creator>
<dc:creator>Torres, D.</dc:creator>
<dc:creator>Jablonski, M.</dc:creator>
<dc:creator>Torres-Martinez, H. H.</dc:creator>
<dc:creator>Martinez, J. L.</dc:creator>
<dc:creator>Hernandez, H. O.</dc:creator>
<dc:creator>Ocelotl, J.</dc:creator>
<dc:creator>Garces, Y.</dc:creator>
<dc:creator>Barchi, M.</dc:creator>
<dc:creator>Dubrovsky, J. G.</dc:creator>
<dc:creator>Darszon, A.</dc:creator>
<dc:creator>Buffone, M.</dc:creator>
<dc:creator>Rodriguez, R.</dc:creator>
<dc:creator>Rendon-Mancha, J. M.</dc:creator>
<dc:creator>Wood, C.</dc:creator>
<dc:creator>Hernandez, A.</dc:creator>
<dc:creator>Krapf, D.</dc:creator>
<dc:creator>Crevenna, A. H.</dc:creator>
<dc:creator>Guerrero, A.</dc:creator>
<dc:date>2021-10-18</dc:date>
<dc:identifier>doi:10.1101/2021.10.17.464398</dc:identifier>
<dc:title><![CDATA[Nanoscopic resolution within a single imaging frame]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.14.496138v1?rss=1">
<title>
<![CDATA[
A choline-releasing glycerophosphodiesterase essential for phosphatidylcholine biosynthesis and blood stage development in the malaria parasite 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.14.496138v1?rss=1"
</link>
<description><![CDATA[
The malaria parasite Plasmodium falciparum synthesizes significant amounts of phospholipids to meet the demands of replication within red blood cells. De novo phosphatidylcholine (PC) biosynthesis via the Kennedy pathway is essential, requiring choline that is primarily sourced from host serum lysophosphatidylcholine (lysoPC). LysoPC also acts as an environmental sensor to regulate parasite sexual differentiation. Despite these critical roles for host lysoPC, the enzyme(s) involved in its breakdown to free choline for PC synthesis are unknown. Here we show that a parasite glycerophosphodiesterase (PfGDPD) is indispensable for blood stage parasite proliferation. Exogenous choline rescues growth of PfGDPD-null parasites, directly linking PfGDPD function to choline incorporation. Loss of PfGDPD reduces choline uptake from lysoPC, resulting in depletion of several PC species in the parasite, whilst purified PfGDPD releases choline from glycerophosphocholine in vitro. Our results identify PfGDPD as a choline-releasing glycerophosphodiesterase that mediates a critical step in PC biosynthesis and parasite survival.
]]></description>
<dc:creator>Ramaprasad, A.</dc:creator>
<dc:creator>Burda, P.-C.</dc:creator>
<dc:creator>Calvani, E.</dc:creator>
<dc:creator>Sait, A.</dc:creator>
<dc:creator>Palma-Duran, S. A.</dc:creator>
<dc:creator>Withers-Martinez, C.</dc:creator>
<dc:creator>Hackett, F.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:creator>Gilberger, T.</dc:creator>
<dc:creator>Blackman, M.</dc:creator>
<dc:date>2022-06-14</dc:date>
<dc:identifier>doi:10.1101/2022.06.14.496138</dc:identifier>
<dc:title><![CDATA[A choline-releasing glycerophosphodiesterase essential for phosphatidylcholine biosynthesis and blood stage development in the malaria parasite]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.14.495937v1?rss=1">
<title>
<![CDATA[
Crowd-sourced benchmarking of single-sample tumour subclonal reconstruction 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.14.495937v1?rss=1"
</link>
<description><![CDATA[
Tumours are dynamically evolving populations of cells. Subclonal reconstruction algorithms use bulk DNA sequencing data to quantify parameters of tumour evolution, allowing assessment of how cancers initiate, progress and respond to selective pressures. A plethora of subclonal reconstruction algorithms have been created, but their relative performance across the varying biological and technical features of real-world cancer genomic data is unclear. We therefore launched the ICGC-TCGA DREAM Somatic Mutation Calling -- Tumour Heterogeneity and Evolution Challenge. This seven-year community effort used cloud-computing to benchmark 31 containerized subclonal reconstruction algorithms on 51 simulated tumours. Each algorithm was scored for accuracy on seven independent tasks, leading to 12,061 total runs. Algorithm choice influenced performance significantly more than tumour features, but purity-adjusted read-depth, copy number state and read mappability were associated with performance of most algorithms on most tasks. No single algorithm was a top performer for all seven tasks and existing ensemble strategies were surprisingly unable to outperform the best individual methods, highlighting a key research need. All containerized methods, evaluation code and datasets are available to support further assessment of the determinants of subclonal reconstruction accuracy and development of improved methods to understand tumour evolution.
]]></description>
<dc:creator>Salcedo, A.</dc:creator>
<dc:creator>Tarabichi, M.</dc:creator>
<dc:creator>Buchanan, A.</dc:creator>
<dc:creator>Espiritu, S. M. G.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Zhu, K.</dc:creator>
<dc:creator>Yang, T.-H. O.</dc:creator>
<dc:creator>Leshchiner, I.</dc:creator>
<dc:creator>Anastassiou, D.</dc:creator>
<dc:creator>Guan, Y.</dc:creator>
<dc:creator>Jang, G. H.</dc:creator>
<dc:creator>Haase, K.</dc:creator>
<dc:creator>Deshwar, A. G.</dc:creator>
<dc:creator>Zou, W.</dc:creator>
<dc:creator>Umar, I.</dc:creator>
<dc:creator>Dentro, S.</dc:creator>
<dc:creator>Wintersinger, J. A.</dc:creator>
<dc:creator>Chiotti, K.</dc:creator>
<dc:creator>Demeulemeester, J.</dc:creator>
<dc:creator>Jolly, C.</dc:creator>
<dc:creator>Scyza, L.</dc:creator>
<dc:creator>Ko, M.</dc:creator>
<dc:creator>PCAWG-11 Working Group,</dc:creator>
<dc:creator>SMC-Het Participants,</dc:creator>
<dc:creator>Wedge, D. C.</dc:creator>
<dc:creator>Morris, Q. D.</dc:creator>
<dc:creator>Ellrot, K.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>Boutros, P. C.</dc:creator>
<dc:date>2022-06-15</dc:date>
<dc:identifier>doi:10.1101/2022.06.14.495937</dc:identifier>
<dc:title><![CDATA[Crowd-sourced benchmarking of single-sample tumour subclonal reconstruction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.13.495933v1?rss=1">
<title>
<![CDATA[
A computationally-enhanced hiCLIP atlas reveals Staufen1 RNA binding features and links 3' UTR structure to RNA metabolism 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.13.495933v1?rss=1"
</link>
<description><![CDATA[
The structure of mRNA molecules plays an important role in its interactions with trans-acting factors, notably RNA binding proteins (RBPs), thus contributing to the functional consequences of this interplay. However, current transcriptome-wide experimental methods to chart these interactions are limited by their poor sensitivity. Here we extend the hiCLIP atlas of duplexes bound by Staufen1 (STAU1) [~]10-fold, through careful consideration of experimental assumptions, and the development of bespoke computational methods which we apply to existing data. We present Tosca, a Nextflow computational pipeline for the processing, analysis and visualisation of proximity ligation sequencing data generally. We use our extended duplex atlas to discover insights into the RNA selectivity of STAU1, revealing the importance of structural symmetry and duplex-span-dependent nucleotide composition. Furthermore, we identify heterogeneity in the relationship between STAU1-bound 3 UTRs and metabolism of the associated RNAs that we relate to RNA structure: transcripts with short-range proximal 3 UTR duplexes have high degradation rates, but those with long-range duplexes have low rates. Overall, our work enables the integrative analysis of proximity ligation data delivering insights into specific features and effects of RBP-RNA structure interactions.
]]></description>
<dc:creator>Chakrabarti, A. M.</dc:creator>
<dc:creator>Iosub, I. A.</dc:creator>
<dc:creator>Lee, F. C. Y.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:date>2022-06-16</dc:date>
<dc:identifier>doi:10.1101/2022.06.13.495933</dc:identifier>
<dc:title><![CDATA[A computationally-enhanced hiCLIP atlas reveals Staufen1 RNA binding features and links 3' UTR structure to RNA metabolism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.20.496792v1?rss=1">
<title>
<![CDATA[
Comprehensive Analysis of CRISPR Base Editing Outcomes for Multimeric Protein 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.20.496792v1?rss=1"
</link>
<description><![CDATA[
Point mutations in the KCNJ13 gene cause an autosomal recessive, childhood blindness, Leber congenital amaurosis (LCA16) due to a loss-of-function Kir7.1 channel. In the present study, we investigated the etiology of LCA16 caused by a KCNJ13 missense mutation (c.431T>C, p.Leu144Pro) and explored the activity of two cytosine base editors mRNAs (CBEs, BE4max-WTCas9, and evoCDA-SpCas9-NG) as a proof-of-concept therapeutic option. We observed the KCNJ13-related retinopathy phenotype in patients harboring L144P mutation. Our in-silico prediction and in vitro validation demonstrated that L144P mutation affects the channel function. We observed high on-target efficiency in the CBEs treated L144P mutant gene expressing HEK-293 cells. Strikingly, our evaluation of base editing efficacy using electrophysiology showed negligible channel function. We found that the editing bystander  Cs in the protospacer region led to a missense change (L143F) in evoCDA edited cells and only silent changes in BE4max edited cells. Upon investigation of the effect of the synonymous codon, our extended analysis revealed distortion of mRNA structure, altered half-life, and/or low abundance of the cognate tRNA. We propose that KCNJ13-L144P mutation or other genes that share similar genetic complexity may be challenging to correct with the current generation of CRISPR base editors, and a combinational therapy using CRISPR base editors with a tighter editing window and requisite cognate-tRNA supplementation could be an alternative therapeutic approach to restore Kir7.1 channel function in LCA16 patients. Other options for hard-to-rescue alleles could employ homology-directed repair using CRISPR/Cas9 nucleases, Prime editing, and AAV-mediated gene augmentation.
]]></description>
<dc:creator>Kabra, M.</dc:creator>
<dc:creator>Moosajee, M.</dc:creator>
<dc:creator>Newby, G.</dc:creator>
<dc:creator>Molugu, K.</dc:creator>
<dc:creator>Saha, K.</dc:creator>
<dc:creator>Liu, D.</dc:creator>
<dc:creator>Pattnaik, B. R.</dc:creator>
<dc:date>2022-06-20</dc:date>
<dc:identifier>doi:10.1101/2022.06.20.496792</dc:identifier>
<dc:title><![CDATA[Comprehensive Analysis of CRISPR Base Editing Outcomes for Multimeric Protein]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.27.497841v1?rss=1">
<title>
<![CDATA[
Craniofacial dysmorphology in Down Syndrome is caused by increased dosage of Dyrk1a and at least three other genes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.27.497841v1?rss=1"
</link>
<description><![CDATA[
Down syndrome (DS), trisomy of human chromosome 21 (Hsa21), occurs in 1 in 800 live births and is the most common human aneuploidy. DS results in multiple phenotypes, including craniofacial dysmorphology, characterised by midfacial hypoplasia, brachycephaly and micrognathia. The genetic and developmental causes of this are poorly understood. Using morphometric analysis of the Dp1Tyb mouse model of DS and an associated genetic mouse genetic mapping panel, we demonstrate that four Hsa21-orthologous regions of mouse chromosome 16 contain dosage-sensitive genes that cause the DS craniofacial phenotype, and identify one of these causative genes as Dyrk1a. We show that the earliest and most severe defects in Dp1Tyb skulls are in bones of neural crest (NC) origin, and that mineralisation of the Dp1Tyb skull base synchondroses is aberrant. Furthermore, we show that increased dosage of Dyrk1a results in decreased NC cell proliferation and a decrease in size and cellularity of the NC-derived frontal bone primordia. Thus, DS craniofacial dysmorphology is caused by increased dosage of Dyrk1a and at least three other genes.

Summary statementCraniofacial dysmorphology in mouse models of Down syndrome is caused by increased dosage of at least four genes including Dyrk1a, resulting in reduced proliferation of neural crest-derived cranial bone progenitors.
]]></description>
<dc:creator>Redhead, Y.</dc:creator>
<dc:creator>Gibbins, D.</dc:creator>
<dc:creator>Lana-Elola, E.</dc:creator>
<dc:creator>Watson-Scales, S.</dc:creator>
<dc:creator>Dobson, L.</dc:creator>
<dc:creator>Krause, M.</dc:creator>
<dc:creator>Liu, K.</dc:creator>
<dc:creator>Fisher, E.</dc:creator>
<dc:creator>Green, J.</dc:creator>
<dc:creator>Tybulewicz, V.</dc:creator>
<dc:date>2022-06-28</dc:date>
<dc:identifier>doi:10.1101/2022.06.27.497841</dc:identifier>
<dc:title><![CDATA[Craniofacial dysmorphology in Down Syndrome is caused by increased dosage of Dyrk1a and at least three other genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.07.01.498418v1?rss=1">
<title>
<![CDATA[
A conserved role of Hippo signaling in initiation of the first lineage specification event across mammals 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.07.01.498418v1?rss=1"
</link>
<description><![CDATA[
Our understanding of the molecular events driving cell specification in early mammalian development relies mainly on mouse studies, and it remains unclear whether these mechanisms are conserved across mammals, including humans. We have recently shown that the establishment of cell polarity via aPKC is a conserved event in the initiation of the trophectoderm (TE) placental program in mouse, cow, and human embryos. However, the molecular mechanisms transducing cell polarity into cell fate in cow and human embryos is unknown. Here, we have examined the evolutionary conservation of the molecular cascade downstream of aPKC in four different mammalian species: mouse, rat, cow, and human. Surprisingly, by morphokinetic and immunofluorescence analyses, we observe that rat embryos more closely recapitulate human and cow developmental dynamics, in comparison to the mouse. Nevertheless, in all four species, inhibition of the Hippo pathway by targeting LATS kinases is sufficient to drive ectopic TE initiation and downregulation of SOX2, a marker of the inner cell mass. Our comparative embryology approach uncovered intriguing differences as well as similarities in a fundamental developmental process among mammals, reinforcing the importance of cross-species investigations.
]]></description>
<dc:creator>Gerri, C.</dc:creator>
<dc:creator>McCarthy, A.</dc:creator>
<dc:creator>Scott, G. M.</dc:creator>
<dc:creator>Regin, M.</dc:creator>
<dc:creator>Brumm, S.</dc:creator>
<dc:creator>Simon, C. S.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Montesinos, C.</dc:creator>
<dc:creator>Hassitt, C.</dc:creator>
<dc:creator>Hockenhull, S.</dc:creator>
<dc:creator>Hampshire, D.</dc:creator>
<dc:creator>Elder, K.</dc:creator>
<dc:creator>Snell, P.</dc:creator>
<dc:creator>Christie, L.</dc:creator>
<dc:creator>Fouladi-Nashta, A. A.</dc:creator>
<dc:creator>Van de Velde, H.</dc:creator>
<dc:creator>Niakan, K.</dc:creator>
<dc:date>2022-07-02</dc:date>
<dc:identifier>doi:10.1101/2022.07.01.498418</dc:identifier>
<dc:title><![CDATA[A conserved role of Hippo signaling in initiation of the first lineage specification event across mammals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.28.497919v1?rss=1">
<title>
<![CDATA[
TRIM7 restricts Coxsackievirus and norovirus infection by detecting the C-terminalglutamine generated by 3C protease processing 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.28.497919v1?rss=1"
</link>
<description><![CDATA[
TRIM7 catalyses the ubiquitination of multiple substrates with unrelated biological functions. This cross-reactivity is at odds with the specificity usually displayed by enzymes, including ubiquitin ligases. Here we show that TRIM7s extreme substrate promiscuity is due to a highly unusual binding mechanism, in which the PRYSPRY domain captures any ligand with a C-terminal helix that terminates in a hydrophobic residue followed by a glutamine. Many of the non-structural proteins found in RNA viruses contain C-terminal glutamines as a result of polyprotein cleavage by 3C protease. This viral processing strategy generates novel substrates for TRIM7 and explains its ability to inhibit Coxsackie virus and norovirus replication. In addition to viral proteins, cellular proteins such as glycogenin have evolved C-termini that make them a TRIM7 substrate. The  helix-{Phi}Q degron motif recognised by TRIM7 is reminiscent of the N-end degron system and is found in [~] 1% of cellular proteins. These features, together with TRIM7s restricted tissue expression and lack of immune regulation suggest that viral restriction may not be its physiological function.
]]></description>
<dc:creator>Luptak, J.</dc:creator>
<dc:creator>Mallery, D.</dc:creator>
<dc:creator>Jahun, A. S.</dc:creator>
<dc:creator>Albecka, A.</dc:creator>
<dc:creator>Clift, D.</dc:creator>
<dc:creator>Ather, O.</dc:creator>
<dc:creator>Slodkowicz, G.</dc:creator>
<dc:creator>Goodfellow, I.</dc:creator>
<dc:creator>James, L.</dc:creator>
<dc:date>2022-07-05</dc:date>
<dc:identifier>doi:10.1101/2022.06.28.497919</dc:identifier>
<dc:title><![CDATA[TRIM7 restricts Coxsackievirus and norovirus infection by detecting the C-terminalglutamine generated by 3C protease processing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.07.05.498824v1?rss=1">
<title>
<![CDATA[
Polymorphisms Predicting Phylogeny in Hepatitis B Virus (HBV) 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.07.05.498824v1?rss=1"
</link>
<description><![CDATA[
Hepatitis B viruses (HBV) are compact viruses with circular genomes of [~]3.2kb in length. Four genes (HBx, Core, Surface and Polymerase) generating seven products are encoded on overlapping reading frames. Ten HBV genotypes have been characterised (A-J), which may account for differences in transmission, outcomes of infection, and treatment response. However, HBV genotyping is rarely undertaken, and sequencing remains inaccessible in many settings. We used a machine learning approach based on random forest algorithms (RFA) to assess which amino acid (aa) sites in the genome are most informative for determining genotype. We downloaded 5496 genome-length HBV sequences from a public database, excluding recombinant sequences, regions with conserved indels, and genotypes I/J. Each gene was separately translated into aa, and the proteins concatenated into a single sequence (length 1614aa). Using RFA, we searched for aa sites predictive of genotype, and assessed co-variation among the sites with a Mutual Information (MI)-based method. We were able to discriminate confidently between genotypes A-H using 10 aa sites. 5/10 sites were identified in Polymerase (Pol), of which 4/5 were in the spacer domain, and a single site in reverse transcriptase. A further 4/10 sites were located in Surface protein, and a single site in HBx. There were no informative sites in Core. Properties of the aa were generally not conserved between genotypes at informative sites. Co-variation analysis identified 55 pairs of highly-linked sites. Three RFA-identified sites were represented across all pairs (two sites in spacer, and one in HBx). Residues that co-vary with these sites are concentrated in the small HBV surface gene. We also observe a cluster of sites adjacent to the Surface promoter region that co-vary with a spacer residue. Overall, we have shown that RFA analysis is a powerful tool for identifying aa sites that predict HBV lineage, with an unexpectedly high number of such sites in the spacer domain, which has conventionally been viewed as unimportant for structure or function. Our results improve ease of genotype prediction from limited regions of HBV sequence, and may have implications for understanding HBV evolution and the role of the spacer domain.
]]></description>
<dc:creator>Lourenco, J.</dc:creator>
<dc:creator>McNaughton, A. L.</dc:creator>
<dc:creator>Pley, C.</dc:creator>
<dc:creator>Obolski, U. L.</dc:creator>
<dc:creator>Gupta, S.</dc:creator>
<dc:creator>Matthews, P. C.</dc:creator>
<dc:date>2022-07-05</dc:date>
<dc:identifier>doi:10.1101/2022.07.05.498824</dc:identifier>
<dc:title><![CDATA[Polymorphisms Predicting Phylogeny in Hepatitis B Virus (HBV)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.07.07.498804v1?rss=1">
<title>
<![CDATA[
Deep proteomic analysis of human microglia and model systems reveal fundamental biological differences of in vitro and ex vivo cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.07.07.498804v1?rss=1"
</link>
<description><![CDATA[
Using high resolution quantitative mass spectrometry, we have generated the most comprehensive human and mouse microglia proteomic datasets to date, consisting of over 11,000 proteins across all six microglia groups. Microglia from different sources share a core protein signature of over 5600 proteins, yet fundamental differences are observed between species and culture conditions, indicating limitations for human disease modelling in mouse or in in vitro cultures of microglia. Mouse ex vivo microglia show important differences at the proteome level such as differential expression of inflammation and Alzheimers Disease associated proteins. We identify a tenfold difference in the protein content of ex vivo and in vitro cells and significant proteome differences associated with protein synthesis, metabolism, microglia marker expression and environmental sensors. Culturing microglia induces rapidly increased growth, protein content and inflammatory protein expression. These changes can be restored by engrafting in vitro cells into the brain, with xenografted hESC-derived microglia closely resembling microglia from human brain. This data provides an important resource for the field and highlights important considerations needed when using model systems to study human physiology and pathology of microglia.
]]></description>
<dc:creator>Lloyd, A. F.</dc:creator>
<dc:creator>Martinez-Muriana, A.</dc:creator>
<dc:creator>Hou, P.</dc:creator>
<dc:creator>Davis, E.</dc:creator>
<dc:creator>Mancuso, R.</dc:creator>
<dc:creator>Brenes-Murillo, A. J.</dc:creator>
<dc:creator>Geric, I.</dc:creator>
<dc:creator>Snellinx, A.</dc:creator>
<dc:creator>Craessaerts, K.</dc:creator>
<dc:creator>Theys, T.</dc:creator>
<dc:creator>Fiers, M.</dc:creator>
<dc:creator>De Strooper, B.</dc:creator>
<dc:creator>Howden, A. J.</dc:creator>
<dc:date>2022-07-07</dc:date>
<dc:identifier>doi:10.1101/2022.07.07.498804</dc:identifier>
<dc:title><![CDATA[Deep proteomic analysis of human microglia and model systems reveal fundamental biological differences of in vitro and ex vivo cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.07.12.499603v1?rss=1">
<title>
<![CDATA[
Enhanced innate immune suppression by SARS-CoV-2 Omicron subvariants BA.4 and BA.5 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.07.12.499603v1?rss=1"
</link>
<description><![CDATA[
SARS-CoV-2 adaptation to humans is evidenced by the emergence of variants of concern (VOCs) with distinct genotypes and phenotypes that facilitate immune escape and enhance transmission frequency. Most recently Omicron subvariants have emerged with heavily mutated spike proteins which facilitate re-infection of immune populations through extensive antibody escape driving replacement of previously-dominant VOCs Alpha and Delta. Interestingly, Omicron is the first VOC to produce distinct subvariants. Here, we demonstrate that later Omicron subvariants, particularly BA.4 and BA.5, have evolved an enhanced capacity to suppress human innate immunity when compared to earliest subvariants BA.1 and BA.2. We find that, like previously dominant VOCs, later Omicron subvariants tend to increase expression of viral innate immune antagonists Orf6 and nucleocapsid. We show Orf6 to be a key contributor to enhanced innate immune suppression during epithelial replication by BA.5 and Alpha, reducing innate immune signaling through IRF3 and STAT1. Convergent VOC evolution of enhanced innate immune antagonist expression suggests common pathways of adaptation to humans and links VOC, and in particular Omicron subvariant, dominance to improved innate immune evasion.
]]></description>
<dc:creator>Reuschl, A.-K.</dc:creator>
<dc:creator>Thorne, L. G.</dc:creator>
<dc:creator>Whelan, M. X. V.</dc:creator>
<dc:creator>Mesner, D.</dc:creator>
<dc:creator>Ragazzini, R.</dc:creator>
<dc:creator>Dowgier, G.</dc:creator>
<dc:creator>Bogoda, N.</dc:creator>
<dc:creator>Turner, J. E.</dc:creator>
<dc:creator>Furnon, W.</dc:creator>
<dc:creator>Cowton, V. M.</dc:creator>
<dc:creator>de Lorenzo, G.</dc:creator>
<dc:creator>Bonfanti, P.</dc:creator>
<dc:creator>Palmarini, M.</dc:creator>
<dc:creator>Patel, A. H.</dc:creator>
<dc:creator>Jolly, C.</dc:creator>
<dc:creator>Towers, G. J.</dc:creator>
<dc:date>2022-07-12</dc:date>
<dc:identifier>doi:10.1101/2022.07.12.499603</dc:identifier>
<dc:title><![CDATA[Enhanced innate immune suppression by SARS-CoV-2 Omicron subvariants BA.4 and BA.5]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.07.12.499640v1?rss=1">
<title>
<![CDATA[
A branching model of cell fate decisions in the enteric nervous system 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.07.12.499640v1?rss=1"
</link>
<description><![CDATA[
How neurogenesis and gliogenesis are coordinated during development and why mature glial cells often share properties with neuroectodermal progenitors remains unclear. Here, we have used single cell RNA sequencing to map the regulatory landscape of neuronal and glial differentiation in the mammalian enteric nervous system (ENS). Our analysis indicates that neurogenic trajectories branch directly from a linear gliogenic axis defined by autonomic neural crest cells adopting sequential states as they progressively lose their strong neurogenic bias and acquire properties of adult enteric glia. We identify gene modules associated with transcriptional programs driving enteric neurogenesis and cell state transitions along the gliogenic axis. By comparing the chromatin accessibility profile of autonomic neural crest and adult enteric glia we provide evidence that the latter maintain an epigenetic memory of their neurogenic past. Finally, we demonstrate that adult enteric glia maintain neurogenic potential and are capable of generating enteric neurons in certain contexts by activating transcriptional programs employed by early ENS progenitors. Our studies uncover a novel configuration of enteric neurogenesis and gliogenesis that enables the coordinate development of ENS lineages and provides a mechanistic explanation for the ability of enteric glia to be functionally integrated into the adult intestine and simultaneously maintain attributes of early ENS progenitors.
]]></description>
<dc:creator>Laddach, A. C.</dc:creator>
<dc:creator>Chng, S. H.</dc:creator>
<dc:creator>Lasrado, R.</dc:creator>
<dc:creator>Progatzky, F.</dc:creator>
<dc:creator>Shapiro, M.</dc:creator>
<dc:creator>Artemov, A.</dc:creator>
<dc:creator>Castaneda, M. S.</dc:creator>
<dc:creator>Erickson, A.</dc:creator>
<dc:creator>Bon-Frauches, A. C.</dc:creator>
<dc:creator>Kleinjung, J.</dc:creator>
<dc:creator>Boeing, S.</dc:creator>
<dc:creator>Ultanir, S.</dc:creator>
<dc:creator>Adameyko, I.</dc:creator>
<dc:creator>Pachnis, V.</dc:creator>
<dc:date>2022-07-13</dc:date>
<dc:identifier>doi:10.1101/2022.07.12.499640</dc:identifier>
<dc:title><![CDATA[A branching model of cell fate decisions in the enteric nervous system]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.07.12.499786v1?rss=1">
<title>
<![CDATA[
OptoPi: An open source flexible platform for the analysis of small animal behaviour 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.07.12.499786v1?rss=1"
</link>
<description><![CDATA[
Behaviour is the ultimate output of neural circuit computations, and therefore its analysis is a cornerstone of neuroscience research. However, every animal and experimental paradigm requires different illumination conditions to capture and, in some cases, manipulate specific behavioural features. This means that researchers often develop, from scratch, their own solutions and experimental set-ups. Here, we present OptoPi, an open source, affordable ([~] {pound}600), behavioural arena with accompanying multi-animal tracking software. The system features highly customisable and reproducible visible and infrared illumination and allows for temporally precise optogenetic stimulation. OptoPi acquires images using a Raspberry Pi camera, features motorised LED-based illumination, Arduino control, as well as spectrum and irradiance monitoring to fine-tune illumination conditions with real time feedback. Our open-source software (BIO) can be used to simultaneously track multiple animals while accurately keeping individual animals identity both in on-line and off-line modes. We demonstrate the functionality of OptoPi by recording and tracking under different illumination conditions the spontaneous behaviour of larval zebrafish as well as adult Drosophila flies and their first instar larvae, an experimental animal that due to its small size and transparency has classically been hard to track. Further, we showcase OptoPis optogenetic capabilities through a series of experiments using transgenic Drosophila larvae.
]]></description>
<dc:creator>Cano-Ferrer, X.</dc:creator>
<dc:creator>Roberts, R. J. V.</dc:creator>
<dc:creator>French, A. S.</dc:creator>
<dc:creator>de Folter, J.</dc:creator>
<dc:creator>Gong, H.</dc:creator>
<dc:creator>Nightingale, L.</dc:creator>
<dc:creator>Strange, A.</dc:creator>
<dc:creator>Imbert, A.</dc:creator>
<dc:creator>Prieto-Godino, L. L.</dc:creator>
<dc:date>2022-07-13</dc:date>
<dc:identifier>doi:10.1101/2022.07.12.499786</dc:identifier>
<dc:title><![CDATA[OptoPi: An open source flexible platform for the analysis of small animal behaviour]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.07.14.498938v1?rss=1">
<title>
<![CDATA[
Robust, single-shot, optical autofocus system utilizing cylindrical lenses to provide high precision and long range of operation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.07.14.498938v1?rss=1"
</link>
<description><![CDATA[
We present a robust, "real-time" optical autofocus system for microscopy that provides high accuracy (<230 nm) and long range ([~]130 {micro}m) with a 1.4 numerical aperture oil immersion objective lens. This autofocus can operate in a closed loop, single-shot functionality over a range of {+/-}37.5 {micro}m and can also operate as a 2-step process up to {+/-}68 {micro}m. A real-time autofocus capability is useful for experiments with long image data acquisition times, including single molecule localization microscopy, that may be impacted by defocusing resulting from drift of components, e.g., due to changes in temperature or mechanical drift. It is also vital for automated slide scanning or multiwell plate imaging where the sample may not be in the same horizontal plane for every field of view during the image data acquisition. To realise high precision and long range, we implement orthogonal optical readouts using cylindrical lenses. We demonstrate the performance of this new optical autofocus system with automated multiwell plate imaging and single molecule localisation microscopy and illustrate the benefit of using a superluminescent diode as the autofocus light source.
]]></description>
<dc:creator>Lightley, J.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Garcia, E.</dc:creator>
<dc:creator>Alexandrov, Y.</dc:creator>
<dc:creator>Dunsby, C.</dc:creator>
<dc:creator>Neil, M. A. A.</dc:creator>
<dc:creator>French, P. M. W.</dc:creator>
<dc:date>2022-07-16</dc:date>
<dc:identifier>doi:10.1101/2022.07.14.498938</dc:identifier>
<dc:title><![CDATA[Robust, single-shot, optical autofocus system utilizing cylindrical lenses to provide high precision and long range of operation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.07.20.500909v1?rss=1">
<title>
<![CDATA[
Phenotypic profile of Mycobacterium tuberculosis-specific CD4 T cell responses in HIV-positive patients who develop Tuberculosis-associated Immune Reconstitution Inflammatory Syndrome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.07.20.500909v1?rss=1"
</link>
<description><![CDATA[
BackgroundTuberculosis-associated immune reconstitution inflammatory syndrome (TB-IRIS) is a frequent complication of co-treatment for TB and HIV-1. We characterized Mtb-specific CD4 T cell phenotype and transcription factor profile associated with the development of TB-IRIS.

MethodsWe examined the role of CD4 T-cell transcription factors in a murine model of mycobacterial IRIS. In humans, we compared longitudinally on antiretroviral therapy (ART) the magnitude, activation, transcription factor profile and cytotoxic potential of Mtb-specific CD4 T cells between TB-IRIS (n=25) and appropriate non-IRIS control patients (n=18) using flow cytometry.

ResultsIn the murine model, CD4 T cell expression of Eomes, but not Tbet, was associated with experimentally induced IRIS. In patients, TB-IRIS onset was associated with the expansion of Mtb-specific IFN{gamma}+CD4 T cells (p=0.039). TB-IRIS patients had higher HLA-DR expression (p=0.016), but no differences in the expression of T-bet or Eomes were observed. At TB-IRIS onset, Eomes+Tbet+Mtb-specific IFN{gamma}+CD4+ T cells showed higher expression of Granzyme B in TB-IRIS patients (p=0.026).

ConclusionWhile the murine model of MAC-IRIS suggests that Eomes+CD4 T cells underly IRIS, TB-IRIS was not associated with Eomes expression in patients. Mtb-specific IFN{gamma}+CD4 T cell responses in TB-IRIS patients are differentiated, highly activated and potentially cytotoxic.
]]></description>
<dc:creator>Moseki, M. R.</dc:creator>
<dc:creator>Barber, D.</dc:creator>
<dc:creator>Du Bruyn, E.</dc:creator>
<dc:creator>Shey, M.</dc:creator>
<dc:creator>Van der Plas, H.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Meintjes, G.</dc:creator>
<dc:creator>Riou, C.</dc:creator>
<dc:date>2022-07-22</dc:date>
<dc:identifier>doi:10.1101/2022.07.20.500909</dc:identifier>
<dc:title><![CDATA[Phenotypic profile of Mycobacterium tuberculosis-specific CD4 T cell responses in HIV-positive patients who develop Tuberculosis-associated Immune Reconstitution Inflammatory Syndrome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.07.21.500989v1?rss=1">
<title>
<![CDATA[
The APE2 nuclease is essential for DNA double strand break repairby microhomology-mediated end-joining. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.07.21.500989v1?rss=1"
</link>
<description><![CDATA[
Microhomology-mediated end-joining (MMEJ) is an intrinsically mutagenic pathway of DNA double strand break repair essential for proliferation of homologous recombination (HR) deficient tumors. While targeting MMEJ has emerged as a powerful strategy to eliminate HR-deficient (HRD) cancers, this is limited by an incomplete understanding of the mechanism and factors required for MMEJ repair. Here, we identify the APE2 nuclease as a novel MMEJ effector. We show that loss of APE2 blocks the fusion of deprotected telomeres by MMEJ and inhibits MMEJ in DNA repair reporter assays to levels comparable to Pol Theta suppression. Mechanistically, we demonstrate that APE2 possesses intrinsic flap-cleaving activity, that its MMEJ function in cells depends on its nuclease domain and further identify uncharacterized domains required for recruitment to damaged DNA. We conclude that HR-deficient cells are addicted to APE2 due to a previously unappreciated role in MMEJ, which could be exploited in the treatment of cancer.
]]></description>
<dc:creator>Fleury, H.</dc:creator>
<dc:creator>MacEachern, M. K.</dc:creator>
<dc:creator>Stiefel, C. M.</dc:creator>
<dc:creator>Anand, R.</dc:creator>
<dc:creator>Sempeck, C.</dc:creator>
<dc:creator>Nebenfuehr, B.</dc:creator>
<dc:creator>Dodd, B.</dc:creator>
<dc:creator>Taylor, E.</dc:creator>
<dc:creator>Dansoko, D.</dc:creator>
<dc:creator>Ortega, R.</dc:creator>
<dc:creator>Leung, J. W.</dc:creator>
<dc:creator>Boulton, S. J.</dc:creator>
<dc:creator>Arnoult, N.</dc:creator>
<dc:date>2022-07-22</dc:date>
<dc:identifier>doi:10.1101/2022.07.21.500989</dc:identifier>
<dc:title><![CDATA[The APE2 nuclease is essential for DNA double strand break repairby microhomology-mediated end-joining.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.07.26.501519v1?rss=1">
<title>
<![CDATA[
Optimal control of gene regulatory networks for morphogen-driven tissue patterning 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.07.26.501519v1?rss=1"
</link>
<description><![CDATA[
The organised generation of functionally distinct cell types in developing tissues depends on establishing spatial patterns of gene expression. In many cases, this is directed by spatially graded chemical signals - known as morphogens. In the influential "French Flag Model", morphogen concentration is proposed to instruct cells to acquire their specific fate. However, this mechanism has been questioned. It is unclear how it produces timely and organised cell-fate decisions, despite the presence of changing morphogen levels, molecular noise and individual variability. Moreover, feedback is present at various levels in developing tissues introducing dynamics to the process that break the link between morphogen concentration, signaling activity and position. Here we develop an alternative approach using optimal control theory to tackle the problem of morphogen-driven patterning. In this framework, intracellular signalling is derived as the control strategy that guides cells to the correct fate while minimizing a combination of signalling levels and the time taken. Applying this approach demonstrates its utility and recovers key properties of the patterning strategies that are found in experimental data. Together, the analysis offers insight into the design principles that produce timely, precise and reproducible morphogen patterning and it provides an alternative framework to the French Flag paradigm for investigating and explaining the control of tissue patterning.
]]></description>
<dc:creator>Pezzotta, A.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:date>2022-07-26</dc:date>
<dc:identifier>doi:10.1101/2022.07.26.501519</dc:identifier>
<dc:title><![CDATA[Optimal control of gene regulatory networks for morphogen-driven tissue patterning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.07.26.501523v1?rss=1">
<title>
<![CDATA[
cloneMap: a R package to visualise clonal heterogeneity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.07.26.501523v1?rss=1"
</link>
<description><![CDATA[
Genomic evolution and clonal heterogeneity in the soma are widespread and are involved in various pathologies with devastating consequences to human health, most prominently in neoplasia. Tools to interrogate and visualise the diversity of clonal populations in malignant or non-malignant tissues are currently lacking. cloneMap is a tool to represent clonal heterogeneity in a sample or set of samples, accounting for the clone size and phylogenetic relationships between clones. We apply cloneMap to several publicly available cancer sequencing datasets to demonstrate its ability to provide biological inferences of clonal heterogeneity. cloneMap is an R package available on github (https://github.com/amf71/cloneMap).
]]></description>
<dc:creator>Frankell, A.</dc:creator>
<dc:creator>Colliver, E.</dc:creator>
<dc:creator>McGranahan, N.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:date>2022-07-27</dc:date>
<dc:identifier>doi:10.1101/2022.07.26.501523</dc:identifier>
<dc:title><![CDATA[cloneMap: a R package to visualise clonal heterogeneity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.08.01.502287v1?rss=1">
<title>
<![CDATA[
How talin allosterically activates vinculin 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.08.01.502287v1?rss=1"
</link>
<description><![CDATA[
The talin-vinculin axis is a key mechanosensing component of cellular focal adhesions. How talin and vinculin respond to forces and regulate one another remains unclear. By combining single molecule magnetic tweezer experiments, Molecular Dynamics simulations, actin bundling assays, and adhesion assembly experiments in live cells, we here discover a two-ways allosteric network within vinculin as a regulator of the talin-vinculin interaction. We directly observe a maturation process of vinculin upon talin binding which reinforces the binding to talin at a rate of 0.03 s-1. This allosteric transition can compete with force-induced dissociation of vinculin from talin only at 7-10 pN. Mimicking the allosteric activation by mutation yields a vinculin molecule that bundles actin and localizes to focal adhesions in a force-independent manner. Hence, the allosteric switch confines talin-vinculin interactions and focal adhesion build-up to intermediate force levels. The  allosteric vinculin mutant is a valuable molecular tool to further dissect the mechanical and biochemical signalling circuits at focal adhesions and elsewhere.
]]></description>
<dc:creator>Franz, F.</dc:creator>
<dc:creator>Tapia-Rojo, R.</dc:creator>
<dc:creator>Winograd-Katz, S. E.</dc:creator>
<dc:creator>Boujemaa-Paterski, R.</dc:creator>
<dc:creator>Li, W.</dc:creator>
<dc:creator>Unger, T.</dc:creator>
<dc:creator>Albeck, S.</dc:creator>
<dc:creator>Aponte-Santamaria, C.</dc:creator>
<dc:creator>Garcia-Manyes, S.</dc:creator>
<dc:creator>Medalia, O.</dc:creator>
<dc:creator>Geiger, B.</dc:creator>
<dc:creator>Grater, F.</dc:creator>
<dc:date>2022-08-02</dc:date>
<dc:identifier>doi:10.1101/2022.08.01.502287</dc:identifier>
<dc:title><![CDATA[How talin allosterically activates vinculin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.08.04.502772v1?rss=1">
<title>
<![CDATA[
Improved memory CD8 T cell response to delayed vaccine boost is associated with a distinct molecular signature. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.08.04.502772v1?rss=1"
</link>
<description><![CDATA[
Although the prime/boost interval can impact vaccine responses, the criteria for deciding its time length are poorly defined. To address this, we examined CD8 T cell responsiveness to boost in a BALB/c mouse model of intramuscular (i.m.) vaccination by priming with HIV-1 gag-encoding Chimpanzee adenovector, and boosting with HIV-1 gag-encoding Modified Vaccinia virus Ankara. We found that boost was more effective at day(d)100 than at d30 post-prime, as evaluated at d45 post-boost by multi-lymphoid organ assessment of gag-specific CD8 T cell frequency, CD62L-expression (as a guide to memory status) and in vivo killing. RNA-sequencing of splenic gag-primed CD8 T cells at d100 revealed a quiescent, but highly responsive signature, that trended toward a central memory (CD62L+) phenotype. Interestingly, gag-specific CD8 T cell frequency selectively diminished in the blood at d100, relative to the spleen, lymph nodes and bone marrow. These results move forward the rational design of prime/boost intervals.
]]></description>
<dc:creator>Natalini, A.</dc:creator>
<dc:creator>Simonetti, S.</dc:creator>
<dc:creator>Favaretto, G.</dc:creator>
<dc:creator>Lucantonio, L.</dc:creator>
<dc:creator>Peruzzi, G.</dc:creator>
<dc:creator>Munoz-Ruiz, M.</dc:creator>
<dc:creator>Kelly, G.</dc:creator>
<dc:creator>Contino, A. M.</dc:creator>
<dc:creator>Sbrocchi, R.</dc:creator>
<dc:creator>Battella, S.</dc:creator>
<dc:creator>Capone, S.</dc:creator>
<dc:creator>Folgori, A.</dc:creator>
<dc:creator>Nicosia, A.</dc:creator>
<dc:creator>Santoni, A.</dc:creator>
<dc:creator>Hayday, A. C.</dc:creator>
<dc:creator>Di Rosa, F.</dc:creator>
<dc:date>2022-08-05</dc:date>
<dc:identifier>doi:10.1101/2022.08.04.502772</dc:identifier>
<dc:title><![CDATA[Improved memory CD8 T cell response to delayed vaccine boost is associated with a distinct molecular signature.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.08.09.503288v1?rss=1">
<title>
<![CDATA[
Lysosomal damage drives mitochondrial proteome remodelling and reprograms macrophage immunometabolism 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.08.09.503288v1?rss=1"
</link>
<description><![CDATA[
Transient lysosomal damage after infection with cytosolic pathogens or silica crystals uptake results in protease leakage. Whether limited leakage of lysosomal contents into the cytosol affects the function of cytoplasmic organelles is unknown. Here, we show that sterile and non-sterile lysosomal damage triggers a cell death independent proteolytic remodelling of the mitochondrial proteome in macrophages. Mitochondrial metabolic reprogramming required lysosomal leakage of Cathepsin B and Cathepsin L and was independent of proteasome degradation and mitophagy. In a mouse model of endomembrane damage, metabolic analysis confirmed that in vivo, live lung macrophages that internalised crystals displayed impaired mitochondrial function and increased glycolytic and lipid metabolism. Single-cell RNA-sequencing analysis of bronchoalveolar lavage revealed that lysosomal damage skewed metabolic and immune responses primarily in CD36+/LIPA+ and Krt79+/Car4+ subsets of alveolar macrophages. Importantly, modulation of macrophage metabolism with 2-Deoxy- d- glucose and oxamate impacted the host response to Mycobacterium tuberculosis (Mtb) infection in an endomembrane damage dependent manner. This work uncovers a new inter-organelle communication pathway, providing a general mechanism by which macrophages undergo mitochondrial metabolic reprograming after endomembrane damage.
]]></description>
<dc:creator>Bussi, C.</dc:creator>
<dc:creator>Heunis, T.</dc:creator>
<dc:creator>Pellegrino, E.</dc:creator>
<dc:creator>Bernard, E. M.</dc:creator>
<dc:creator>Bah, N.</dc:creator>
<dc:creator>Silva Dos Santos, M.</dc:creator>
<dc:creator>Santucci, P.</dc:creator>
<dc:creator>Aylan, B.</dc:creator>
<dc:creator>Rodgers, A.</dc:creator>
<dc:creator>Fearns, A.</dc:creator>
<dc:creator>Mitschke, J.</dc:creator>
<dc:creator>Moore, C.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Greco, M.</dc:creator>
<dc:creator>Reinheckel, T.</dc:creator>
<dc:creator>Trost, M.</dc:creator>
<dc:creator>Gutierrez, M. G.</dc:creator>
<dc:date>2022-08-09</dc:date>
<dc:identifier>doi:10.1101/2022.08.09.503288</dc:identifier>
<dc:title><![CDATA[Lysosomal damage drives mitochondrial proteome remodelling and reprograms macrophage immunometabolism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.12.495791v1?rss=1">
<title>
<![CDATA[
Neural plate progenitors give rise to both anterior and posterior pituitary cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.12.495791v1?rss=1"
</link>
<description><![CDATA[
The pituitary is the master neuroendocrine gland, which regulates body homeostasis. It consists of the anterior pituitary/adenohypophysis (AH), which harbors hormones producing cells and the posterior pituitary/neurohypophysis (NH), which relays the direct passage of hormones from the brain to the periphery. It is widely accepted that the AH originates from the oral ectoderm (Rathkes pouch) whereas the neural ectoderm contributes to the NH. Using single cell transcriptomics of the zebrafish pituitary we characterized cyp26b1-positive pituicyte of the NH and prop1-positive adenohypophyseal progenitors. We found that these cell types expressed common markers implying lineage relatedness. Genetic tracing revealed that in contrast to the prevailing dogma, neural plate precursors of both zebrafish (her4.3+) and mouse (Sox1+) contribute to both the neurohypophyseal and adenohypophyseal cells. We further show that pituicytes and prop1+ progenitors reside in close anatomical proximity and pituicyte-derived RA-degrading enzyme Cyp26b1 fine-tunes differentiation of prop1+ progenitors into hormone-producing cells. These results challenge the notion that AH cells are exclusively derived from non-neural ectoderm and demonstrate that a cross-talk between neuro- and adeno-hypophyseal cells fine-tunes the development of pituitary neuroendocrine cells.
]]></description>
<dc:creator>Chen, Q.</dc:creator>
<dc:creator>Leshkowitz, D.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>van Impel, A.</dc:creator>
<dc:creator>Schulte-Merker, S.</dc:creator>
<dc:creator>Amit, I.</dc:creator>
<dc:creator>Levkowitz, G.</dc:creator>
<dc:date>2022-06-12</dc:date>
<dc:identifier>doi:10.1101/2022.06.12.495791</dc:identifier>
<dc:title><![CDATA[Neural plate progenitors give rise to both anterior and posterior pituitary cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.08.09.503398v1?rss=1">
<title>
<![CDATA[
Sequential roles for red blood cell binding proteins enable phased commitment to invasion for malaria parasites 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.08.09.503398v1?rss=1"
</link>
<description><![CDATA[
Invasion of red blood cells (RBCs) by Plasmodium merozoites is critical to their continued survival within the host. Two major protein families, the Duffy binding-like proteins (DBPs/EBAs) and the reticulocyte binding like proteins (RBLs/RHs) have been studied extensively in P. falciparum and are hypothesized to have overlapping, but critical roles just prior to host cell entry. The zoonotic malaria parasite, P. knowlesi, has larger invasive merozoites and contains a smaller, less redundant, DBP and RBL repertoire than P. falciparum. One DBP (DBP) and one RBL, normocyte binding protein Xa (NBPXa) are essential for invasion of human RBCs. Taking advantage of the unique biological features of P. knowlesi and iterative CRISPR-Cas9 genome editing, we determine the precise order of key invasion milestones and demonstrate distinct roles for each family. These distinct roles support a mechanism for phased commitment to invasion and can be targeted synergistically with invasion inhibitory antibodies.
]]></description>
<dc:creator>Hart, M. N.</dc:creator>
<dc:creator>Mohring, F.</dc:creator>
<dc:creator>Donvito, S. M.</dc:creator>
<dc:creator>Thomas, J. A.</dc:creator>
<dc:creator>Muller-Sienerth, N.</dc:creator>
<dc:creator>Wright, G. J.</dc:creator>
<dc:creator>Knuepfer, E.</dc:creator>
<dc:creator>Saibil, H. R.</dc:creator>
<dc:creator>Moon, R. W.</dc:creator>
<dc:date>2022-08-11</dc:date>
<dc:identifier>doi:10.1101/2022.08.09.503398</dc:identifier>
<dc:title><![CDATA[Sequential roles for red blood cell binding proteins enable phased commitment to invasion for malaria parasites]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.08.11.503617v1?rss=1">
<title>
<![CDATA[
Cryo-ET of infected cells reveals that a succession of two lattices drives vaccinia virus assembly 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.08.11.503617v1?rss=1"
</link>
<description><![CDATA[
During its cytoplasmic replication, vaccinia virus assembles non-infectious spherical immature virions (IV) coated by a viral D13 lattice. Subsequently, IV mature into infectious brick-shaped intracellular mature virus (IMV) that lack D13. Here, we performed cryo-electron tomography of frozen-hydrated vaccinia-infected cells to structurally characterise the maturation process in situ. During IMV formation a new viral core forms inside IV with a wall consisting of trimeric pillars arranged in a new pseudohexagonal lattice. This lattice appears as a palisade in cross-section. During maturation, which involves a 50% reduction in virion volume, the viral membrane becomes corrugated as it adapts to the newly formed viral core in a process that does not appear to require membrane removal. Our study suggests that the length of this core is determined by the D13 lattice and that the consecutive D13 and palisade lattices control virion shape and dimensions during vaccinia assembly and maturation.
]]></description>
<dc:creator>Hernandez-Gonzalez, M.</dc:creator>
<dc:creator>Calcraft, T.</dc:creator>
<dc:creator>Nans, A.</dc:creator>
<dc:creator>Rosenthal, P. B.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:date>2022-08-12</dc:date>
<dc:identifier>doi:10.1101/2022.08.11.503617</dc:identifier>
<dc:title><![CDATA[Cryo-ET of infected cells reveals that a succession of two lattices drives vaccinia virus assembly]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.08.09.503427v1?rss=1">
<title>
<![CDATA[
The cosmopolitan appendicularian Oikopleura dioica reveals hidden genetic diversity around the globe 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.08.09.503427v1?rss=1"
</link>
<description><![CDATA[
Appendicularian tunicates are some of the most abundant mesozooplankton organisms with key roles in marine trophic webs and global carbon flux. Like most appendicularians with cosmopolitan distributions, Oikopleura dioica Fol, 1872 is considered a single species worldwide based on morphological features that distinguish them from other appendicularians. Despite their abundance however, there are still only [~]70 described appendicularian species, compared with over 2,800 ascidian tunicates. Here we perform a molecular phylogenetic, morphological, and reproductive assessment of O. dioica specimens collected from the Ryukyu Archipelago, mainland Japan, and Europe. The specimens are morphologically very similar, with only detailed examination of the oikoplastic epithelium and quantitative measurements revealing minor distinguishing characteristics. Phylogenetic analyses of the ribosomal gene loci and mitochondrial cytochrome oxidase I (COI) gene strongly indicate that they form three separate genetic clades despite their morphological similarities. Finally, in vitro crosses between the Ryukyu and mainland Japanese specimens show total prezygotic reproductive isolation. Our results reveal that the current taxonomic O. dioica classification likely hides multiple cryptic species, highlighting the genetic diversity and complexity of their population structures. Cryptic organisms are often hidden under a single species name because their morphological similarities make them difficult to disinguish and their correct identification is fundamental to understanding Earths biodiversity. O. dioica is an attractive model to understand how morphological conservation can be maintained despite pronounced genetic divergence.
]]></description>
<dc:creator>Masunaga, A.</dc:creator>
<dc:creator>Mansfield, M. J.</dc:creator>
<dc:creator>Tan, Y.</dc:creator>
<dc:creator>Liu, A. W.</dc:creator>
<dc:creator>Bliznina, A.</dc:creator>
<dc:creator>Barzaghi, P.</dc:creator>
<dc:creator>Hodgetts, T. L.</dc:creator>
<dc:creator>Ferrandez-Roldan, A.</dc:creator>
<dc:creator>Canestro, C.</dc:creator>
<dc:creator>Onuma, T.</dc:creator>
<dc:creator>Plessy, C.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:date>2022-08-11</dc:date>
<dc:identifier>doi:10.1101/2022.08.09.503427</dc:identifier>
<dc:title><![CDATA[The cosmopolitan appendicularian Oikopleura dioica reveals hidden genetic diversity around the globe]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.09.04.506531v1?rss=1">
<title>
<![CDATA[
RTEL-1 and DNA polymerase theta promote subtelomeric DNA synthesis and telomere fusion in C. elegans 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.09.04.506531v1?rss=1"
</link>
<description><![CDATA[
Interstitial telomere sequences (ITS) are degenerate telomere tracts scattered along chromosome arms whose functions are not well understood. We found that critically shortened telomeres of C. elegans telomerase mutants initiate DNA synthesis within ITS tracts that were close to or far from a telomere. Some ITS tracts were targeted recurrently. RTEL-1 dismantles T-loops and recombination intermediates, and DNA polymerase theta (POLQ-1) promotes end-joining using short segments of microhomology. In telomerase mutants, RTEL-1 and POLQ-1 promoted telomere fusion and DNA synthesis at subtelomeric ITS tracts. RTEL-1 is known to suppress homologous recombination, and we found that RTEL-1 similarly suppressed POLQ-1-mediated double-strand break repair. Mutation signatures characteristic of repair by POLQ-1 occurred during initiation of subtelomeric DNA synthesis and at subsequent template shifting events. We propose that RTEL-1 and POLQ-1 play distinct essential roles in subtelomeric DNA synthesis, a process that may contribute significantly to telomere fusion and tumor genome evolution.
]]></description>
<dc:creator>Ahmed, S.</dc:creator>
<dc:creator>Lister-Shimauchi, E.</dc:creator>
<dc:creator>Brady, M.</dc:creator>
<dc:creator>Frenk, S.</dc:creator>
<dc:creator>Harris, B.</dc:creator>
<dc:creator>Ortiz, A.-M. L.</dc:creator>
<dc:creator>Memili, A.</dc:creator>
<dc:creator>Nguyen, M.</dc:creator>
<dc:creator>Boulton, S.</dc:creator>
<dc:date>2022-09-04</dc:date>
<dc:identifier>doi:10.1101/2022.09.04.506531</dc:identifier>
<dc:title><![CDATA[RTEL-1 and DNA polymerase theta promote subtelomeric DNA synthesis and telomere fusion in C. elegans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.09.05.506481v1?rss=1">
<title>
<![CDATA[
Hunter-gatherer admixture facilitated natural selection in Neolithic European farmers 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.09.05.506481v1?rss=1"
</link>
<description><![CDATA[
Ancient DNA has revealed multiple episodes of admixture in human prehistory during geographic expansions associated with cultural innovations. One important example is the expansion of Neolithic agricultural groups out of the Near East into Europe, and their consequent admixture with Mesolithic hunter-gatherers. Ancient genomes from this period provide an opportunity to study the role of admixture in providing new genetic variation for selection to act upon, and also to identify genomic regions that resisted hunter-gatherer introgression and may thus contribute to agricultural adaptations. We used genome-wide DNA from 728 individuals spanning Mesolithic and Neolithic Europe to infer ancestry deviations in the genomes of admixed individuals, and to test for natural selection after admixture using a new method based on testing for deviations from a genome-wide null distribution. We find that the region around the pigmentation-associated gene SLC24A5 shows the greatest overrepresentation of Neolithic ancestry in the genome (|Z| = 3.45). In contrast, we find the greatest overrepresentation of Mesolithic local ancestry across the key immunity locus that is the Major Histocompatibility Complex (MHC; |Z| > 4) which also shows allele frequency deviations indicative of a selective sweep following admixture (p =1x10-29). This could reflect negative frequency dependent selection on MHC alleles common in Neolithic populations, or that Mesolithic alleles were positively selected for and facilitated adaptation by Neolithic populations to pathogens, new diets, or other environmental factors. Our results extend previous results that highlight immune function and pigmentation as targets of adaptation in more recent populations to selection processes in the Stone Age, and demonstrate that admixture facilitated selection by contributing new genetic variation.
]]></description>
<dc:creator>Davy, T.</dc:creator>
<dc:creator>Ju, D.</dc:creator>
<dc:creator>Mathieson, I.</dc:creator>
<dc:creator>Skoglund, P.</dc:creator>
<dc:date>2022-09-06</dc:date>
<dc:identifier>doi:10.1101/2022.09.05.506481</dc:identifier>
<dc:title><![CDATA[Hunter-gatherer admixture facilitated natural selection in Neolithic European farmers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.09.05.506621v1?rss=1">
<title>
<![CDATA[
Design principles for selective polarization of PAR proteins by cortical flows 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.09.05.506621v1?rss=1"
</link>
<description><![CDATA[
Clustering of membrane-associated molecules is thought to promote interactions with the actomyosin cortex, enabling size-dependent transport by actin flows. Consistent with this model, in the C. elegans zygote, anterior segregation of the polarity protein PAR-3 requires oligomerization. However, through direct assessment of advection of PAR proteins, we not only find no links between PAR-3 advection and oligomer size, but also observe efficient advection of both anterior and posterior PAR proteins. Consequently, differential cortex engagement cannot account for selective size-dependent PAR protein transport. Instead, combining experiment and theory we demonstrate that segregation efficiency of PAR proteins by cortical flow is determined by the stability of membrane association, which is enhanced by clustering and specifies persistence of transport. Indeed, stabilizing membrane association was sufficient to invert polarity of a normally posterior PAR protein. Our data therefore indicate that advection of membrane-associated proteins is more pervasive than anticipated and thus cells must tune membrane association dynamics to achieve differential transport by cortical flows.
]]></description>
<dc:creator>Illukkumbura, R.</dc:creator>
<dc:creator>Hirani, N.</dc:creator>
<dc:creator>Borrego-Pinto, J.</dc:creator>
<dc:creator>Bland, T.</dc:creator>
<dc:creator>Ng, K.</dc:creator>
<dc:creator>Hubatsch, L.</dc:creator>
<dc:creator>McQuade, J.</dc:creator>
<dc:creator>Endres, R.</dc:creator>
<dc:creator>Goehring, N.</dc:creator>
<dc:date>2022-09-06</dc:date>
<dc:identifier>doi:10.1101/2022.09.05.506621</dc:identifier>
<dc:title><![CDATA[Design principles for selective polarization of PAR proteins by cortical flows]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.09.06.506650v1?rss=1">
<title>
<![CDATA[
Long-term retention of antigens in germinal centres is controlled by the spatial organisation of the follicular dendritic cell network 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.09.06.506650v1?rss=1"
</link>
<description><![CDATA[
Germinal centers (GCs) require sustained availability of antigens to promote antibody affinity maturation against pathogens and vaccines. A key source of antigens for GC B cells are immune complexes (ICs) displayed on follicular dendritic cells (FDCs). Here we show that FDC spatial organization regulates antigen dynamics in the GC. We show the existence of two light zone (LZ) FDC populations, which differ in the duration of antigen retention. While the entire light zone (LZ) FDC network captures ICs initially, only the central cells of the network function as a long-term antigen reservoir, where different antigens arriving from subsequent immunizations co-localize. Mechanistically, central FDCs constitutively express subtly higher CR2 membrane densities than peripheral FDCs, which strongly increases the IC retention half-life. Even though repeated immunizations gradually saturate central FDCs, B cell responses remain efficient because new antigens partially displace old ones. These results reveal the principles shaping antigen display on FDCs during the GC reaction.
]]></description>
<dc:creator>Martinez-Riano, A.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Boeing, S.</dc:creator>
<dc:creator>Minoughan, S.</dc:creator>
<dc:creator>Casal, A.</dc:creator>
<dc:creator>Spillane, K. M.</dc:creator>
<dc:creator>Ludewig, B.</dc:creator>
<dc:creator>Tolar, P.</dc:creator>
<dc:date>2022-09-07</dc:date>
<dc:identifier>doi:10.1101/2022.09.06.506650</dc:identifier>
<dc:title><![CDATA[Long-term retention of antigens in germinal centres is controlled by the spatial organisation of the follicular dendritic cell network]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.09.09.507316v1?rss=1">
<title>
<![CDATA[
Dual role of brain endothelial Gpr126 in blood-brain barrier development and ischemic stroke 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.09.09.507316v1?rss=1"
</link>
<description><![CDATA[
The blood-brain barrier (BBB) acquires unique properties for regulation of the neuronal function during development. The genesis of the BBB coupled with angiogenesis is orchestrated by the Wnt/{beta}-catenin signaling pathway. Aside from the importance of Wnt/{beta}-catenin signaling, the molecular mechanisms that regulate these processes are poorly understood. Here, we identify the brain endothelial adhesion G-protein-coupled receptor Gpr126 as a novel target gene of Wnt/{beta}-catenin signaling that is required for postnatal BBB development, and its expression is detrimental for ischemic stroke in adults. We show that Gpr126 expression is high in mouse brain endothelium during BBB formation, but decreases in the adult. Inactivation of Gpr126 in postnatal endothelial cells results in vessel enlargement and impairs acquisition of the BBB characteristics, such as increased neurovascular permeability, and reduced basement membrane protein deposition and pericyte coverage. Mechanistically, Gpr126 is required during developmental angiogenesis to promote endothelial cell migration, acting via an interaction between Lrp1 and 3{beta}1-integrin, which couples vessel morphogenesis to BBB formation. Interestingly, in adult mice with an established BBB, the lack of Gpr126 expression in acute ischemic stroke is protective and coupled with reduced microglia activation, which contributes to an improved neurological outcome. These data identify Gpr126 as a promising therapeutic target to treat ischemic stroke.
]]></description>
<dc:creator>Kakogiannos, N.</dc:creator>
<dc:creator>Scalise, A. A.</dc:creator>
<dc:creator>Martini, E.</dc:creator>
<dc:creator>Maderna, C.</dc:creator>
<dc:creator>Magni, S.</dc:creator>
<dc:creator>Gullotta, G. S.</dc:creator>
<dc:creator>Lampugnani, M. G.</dc:creator>
<dc:creator>Iannelli, F.</dc:creator>
<dc:creator>Beznusenko, G.</dc:creator>
<dc:creator>Mironov, A.</dc:creator>
<dc:creator>Cerutti, C.</dc:creator>
<dc:creator>Bentley, K.</dc:creator>
<dc:creator>Philippides, A.</dc:creator>
<dc:creator>Zanardi, F.</dc:creator>
<dc:creator>Bacigaluppi, M.</dc:creator>
<dc:creator>Martino, G.</dc:creator>
<dc:creator>Dejana, E.</dc:creator>
<dc:creator>Giannotta, M.</dc:creator>
<dc:date>2022-09-10</dc:date>
<dc:identifier>doi:10.1101/2022.09.09.507316</dc:identifier>
<dc:title><![CDATA[Dual role of brain endothelial Gpr126 in blood-brain barrier development and ischemic stroke]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.09.15.508094v1?rss=1">
<title>
<![CDATA[
Sister chromatid cohesion establishment during DNA replication termination 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.09.15.508094v1?rss=1"
</link>
<description><![CDATA[
The cohesin complex tethers sister chromatids together from the moment they are generated in S-phase until their separation in anaphase1,2. This fundamental phenomenon, called sister chromatid cohesion, underpins orderly chromosome segregation. The replisome complex coordinates cohesion establishment with replication of parental DNA3. Cohesion can be established by cohesin complexes bound to DNA before replication4,5, but how replisome interaction with pre-loaded cohesin complexes results in cohesion is not known. Prevailing models suggest cohesion is established by replisome passage through the cohesin ring or by transfer of cohesin behind the replication fork by replisome components5. Unexpectedly, by visualising single replication forks colliding with pre-loaded cohesin complexes, we find that cohesin is pushed by the replisome to where a converging replisome is met. Whilst the converging replisomes are removed during DNA replication termination, cohesin remains on nascent DNA. We demonstrate that these cohesin molecules tether the newly replicated sister DNAs together. Our results support a new model where sister chromatid cohesion is established during DNA replication termination, providing important insight into the molecular mechanism of cohesion establishment.
]]></description>
<dc:creator>Cameron, G.</dc:creator>
<dc:creator>Gruszka, D.</dc:creator>
<dc:creator>Xie, S.</dc:creator>
<dc:creator>Nasmyth, K. A.</dc:creator>
<dc:creator>Srinivasan, M.</dc:creator>
<dc:creator>Yardimci, H.</dc:creator>
<dc:date>2022-09-16</dc:date>
<dc:identifier>doi:10.1101/2022.09.15.508094</dc:identifier>
<dc:title><![CDATA[Sister chromatid cohesion establishment during DNA replication termination]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.09.15.508093v1?rss=1">
<title>
<![CDATA[
O-Linked Sialoglycans Modulate the Proteolysis of SARS-CoV-2 Spike and Contribute to the Mutational Trajectory in Variants of Concern 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.09.15.508093v1?rss=1"
</link>
<description><![CDATA[
The emergence of a polybasic cleavage motif for the protease furin in the SARS-CoV-2 spike protein has been established as a major factor for enhanced viral transmission in humans. The peptide region N-terminal to that motif is extensively mutated in major variants of concern including Alpha, Delta and Omicron. Besides furin, spike proteins from these variants appear to rely on other proteases for maturation, including TMPRSS2 that may share the same cleavage motif. Glycans found near the cleavage site have raised questions about proteolytic processing and the consequences of variant-borne mutations. Here, with a suite of chemical tools, we establish O-linked glycosylation as a major determinant of SARS-CoV-2 spike cleavage by the host proteases furin and TMPRSS2, and as a likely driving force for the emergence of common mutations in variants of concern. We provide direct evidence that the glycosyltransferase GalNAc-T1 primes glycosylation at Thr678 in the living cell, and this glycosylation event is suppressed by many, but not all variant mutations. A novel strategy for rapid bioorthogonal modification of Thr678-containing glycopeptides revealed that introduction of a negative charge completely abrogates furin activity. In a panel of synthetic glycopeptides containing elaborated O-glycans, we found that the sole incorporation of N-acetylgalactosamine did not substantially impact furin activity, but the presence of sialic acid in elaborated O-glycans reduced furin rate by up to 65%. Similarly, O-glycosylation with a sialylated trisaccharide had a negative impact on spike cleavage by TMPRSS2. With a chemistry-centered approach, we firmly establish O-glycosylation as a major determinant of spike maturation and propose that a disruption of O-GalNAc glycosylation is a substantial driving force for the evolution of variants of concern.



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]]></description>
<dc:creator>Gonzalez-Rodriguez, E.</dc:creator>
<dc:creator>Zol-Hanlon, M.</dc:creator>
<dc:creator>Bineva-Todd, G.</dc:creator>
<dc:creator>Marchesi, A.</dc:creator>
<dc:creator>Skehel, M.</dc:creator>
<dc:creator>Mahoney, K.</dc:creator>
<dc:creator>Roustan, C.</dc:creator>
<dc:creator>Borg, A.</dc:creator>
<dc:creator>Di Vagno, L.</dc:creator>
<dc:creator>Kjaer, S.</dc:creator>
<dc:creator>Wrobel, A. G.</dc:creator>
<dc:creator>Benton, D. J.</dc:creator>
<dc:creator>Nawrath, P.</dc:creator>
<dc:creator>Flitsch, S. L.</dc:creator>
<dc:creator>Joshi, D.</dc:creator>
<dc:creator>Gonzalez-Ramirez, A. M.</dc:creator>
<dc:creator>Wilkinson, K. A.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Hurtado-Guerrero, R.</dc:creator>
<dc:creator>Malaker, S. A.</dc:creator>
<dc:creator>Schumann, B.</dc:creator>
<dc:date>2022-09-15</dc:date>
<dc:identifier>doi:10.1101/2022.09.15.508093</dc:identifier>
<dc:title><![CDATA[O-Linked Sialoglycans Modulate the Proteolysis of SARS-CoV-2 Spike and Contribute to the Mutational Trajectory in Variants of Concern]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.09.16.508290v1?rss=1">
<title>
<![CDATA[
Towards In-Silico CLIP-seq: Predicting Protein-RNA Interaction via Sequence-to-Signal Learning 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.09.16.508290v1?rss=1"
</link>
<description><![CDATA[
AO_SCPLOWBSTRACTC_SCPLOWUnraveling sequence determinants which drive protein-RNA interaction is crucial for studying binding mechanisms and the impact of genomic variants. While CLIP-seq allows for transcriptome-wide profiling of in vivo protein-RNA interactions, it is limited to expressed transcripts, requiring computational imputation of missing binding information. Existing classification-based methods predict binding with low resolution and depend on prior labeling of transcriptome regions for training. We present RBPNet, a novel deep learning method, which predicts CLIP crosslink count distribution from RNA sequence at single-nucleotide resolution. By training on up to a million regions, RBPNet achieves high generalization on eCLIP, iCLIP and miCLIP assays, outperforming state-of-the-art classifiers. CLIP-seq suffers from various technical biases, complicating downstream interpretation. RBPNet performs bias correction by modeling the raw signal as a mixture of the protein-specific and background signal. Through model interrogation via Integrated Gradients, RBPNet identifies predictive sub-sequences corresponding to known binding motifs and enables variant-impact scoring via in silico mutagenesis. Together, RBPNet improves inference of protein-RNA interaction, as well as mechanistic interpretation of predictions.
]]></description>
<dc:creator>Horlacher, M.</dc:creator>
<dc:creator>Wagner, N.</dc:creator>
<dc:creator>Moyon, L.</dc:creator>
<dc:creator>Kuret, K.</dc:creator>
<dc:creator>Goedert, N.</dc:creator>
<dc:creator>Salvatore, M.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Gagneur, J.</dc:creator>
<dc:creator>Winther, O.</dc:creator>
<dc:creator>Marsico, A.</dc:creator>
<dc:date>2022-09-19</dc:date>
<dc:identifier>doi:10.1101/2022.09.16.508290</dc:identifier>
<dc:title><![CDATA[Towards In-Silico CLIP-seq: Predicting Protein-RNA Interaction via Sequence-to-Signal Learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.09.22.508986v1?rss=1">
<title>
<![CDATA[
USP7 inactivation suppresses APC-mutant intestinal hyperproliferation and tumor development 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.09.22.508986v1?rss=1"
</link>
<description><![CDATA[
Truncating mutation of the tumor suppressor gene adenomatous polyposis coli (APC) is the hallmark of colorectal cancer (CRC), resulting in constitutive WNT activation. Despite decades of research, targeting WNT signaling in cancer remains challenging due to its essential role in normal stem cell maintenance. We have previously shown that the deubiquitinating enzyme USP7 is a tumor-specific WNT activator in APC-truncated cells by deubiquitinating and stabilizing {beta}-catenin, but its role in gut tumorigenesis is unknown. Here we show in vivo that deletion of Usp7 in Apc-truncated mice inhibits crypt hyperproliferation and intestinal tumor development. Importantly, intestine-specific Usp7 mutation does not yield any phenotype in wildtype animals, indicating that its loss is well tolerated. Unexpectedly, prolonged deletion of Usp7 in Apc+/- intestine induces varying degrees of colitis. Treatment with a USP7 inhibitor suppresses growth of patient-derived cancer organoids in vitro and of xenografts carrying APC truncations. We propose that USP7 inhibition may be efficacious for tumor-specific therapy of sporadic APC-mutated CRC, while patients with germline APC mutations should not receive such treatment.

HighlightsO_LIUsp7 deletion in Apc-truncated mice reduces intestinal tumor development.
C_LIO_LIIntestine-specific Usp7 mutation mutation has no phenotype in wildtype animals.
C_LIO_LITreatment with Usp7 inhibitor suppresses growth of patient-derived cancer organoids carrying Apc truncations in vitro and of xenografts.
C_LI
]]></description>
<dc:creator>Novellasdemunt, L.</dc:creator>
<dc:creator>Kucharska, A.</dc:creator>
<dc:creator>Baulies, A.</dc:creator>
<dc:creator>Vlachogiannis, G.</dc:creator>
<dc:creator>Repana, D.</dc:creator>
<dc:creator>Rowan, A.</dc:creator>
<dc:creator>Suarez-Bonnet, A.</dc:creator>
<dc:creator>Ciccarelli, F.</dc:creator>
<dc:creator>Valeri, N.</dc:creator>
<dc:creator>Li, V. S. W.</dc:creator>
<dc:date>2022-09-23</dc:date>
<dc:identifier>doi:10.1101/2022.09.22.508986</dc:identifier>
<dc:title><![CDATA[USP7 inactivation suppresses APC-mutant intestinal hyperproliferation and tumor development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.09.23.509139v1?rss=1">
<title>
<![CDATA[
Plzf mediates a switch between Fgf signalling regimes in the developing hindbrain 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.09.23.509139v1?rss=1"
</link>
<description><![CDATA[
Developing tissues are sequentially patterned by extracellular signals that are turned on and off at specific times. In the zebrafish hindbrain, fibroblast growth factor (Fgf) signalling has different roles at different developmental stages: in the early hindbrain, transient Fgf3 and Fgf8 signalling from rhombomere 4 is required for correct segmentation, whereas later, neuronal Fgf20 expression confines neurogenesis to specific spatial domains within each rhombomere. How the switch between these two signalling regimes is coordinated is not known. We present evidence that the promyelocytic leukaemia zinc finger (Plzf) transcription factor is required for this transition to happen in an orderly fashion. Plzf expression is high in the early anterior hindbrain, then gradually upregulated posteriorly and confined to neural progenitors. In mutants lacking functional Plzf, fgf3 expression fails to be downregulated and persists until a late stage, resulting in excess and more widespread Fgf signalling during neurogenesis. Accordingly, the spatial pattern of neurogenesis is disrupted in plzf mutants. Our results reveal how the distinct stage-specific roles of Fgf signalling are coordinated in the zebrafish hindbrain.
]]></description>
<dc:creator>Leino, S.</dc:creator>
<dc:creator>Constable, S. C. J.</dc:creator>
<dc:creator>Streit, A.</dc:creator>
<dc:creator>Wilkinson, D. G.</dc:creator>
<dc:date>2022-09-23</dc:date>
<dc:identifier>doi:10.1101/2022.09.23.509139</dc:identifier>
<dc:title><![CDATA[Plzf mediates a switch between Fgf signalling regimes in the developing hindbrain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.09.23.509153v1?rss=1">
<title>
<![CDATA[
Resolving chaperone-assisted protein folding on the ribosome at the peptide level 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.09.23.509153v1?rss=1"
</link>
<description><![CDATA[
The cellular environment is critical for efficient protein maturation, but how proteins fold during biogenesis remains poorly understood. We used hydrogen/deuterium exchange (HDX) mass spectrometry (MS) to define, at peptide resolution, the cotranslational chaperone-assisted folding pathway of Escherichia coli dihydrofolate reductase. On the ribosome, the nascent polypeptide folds via structured intermediates not populated during refolding from denaturant. Association with the ribosome allows these intermediates to form, as otherwise destabilizing C-terminal sequences remain confined in the ribosome exit tunnel. We find that partially-folded nascent chains recruit the chaperone Trigger factor, which uses a large composite hydrophobic/hydrophilic interface to engage folding intermediates without disrupting their structure. In addition, we comprehensively mapped dynamic interactions between the nascent chain and ribosomal proteins, tracing the path of the emerging polypeptide during synthesis. Our work provides a high-resolution description of de novo protein folding dynamics, thereby revealing new mechanisms by which cellular factors shape the conformational search for the native state.
]]></description>
<dc:creator>Wales, T. E.</dc:creator>
<dc:creator>Pajak, A.</dc:creator>
<dc:creator>Roeselova, A.</dc:creator>
<dc:creator>Shivakumaraswamy, S.</dc:creator>
<dc:creator>Howell, S.</dc:creator>
<dc:creator>Hartl, F.-U.</dc:creator>
<dc:creator>Engen, J. R.</dc:creator>
<dc:creator>Balchin, D.</dc:creator>
<dc:date>2022-09-23</dc:date>
<dc:identifier>doi:10.1101/2022.09.23.509153</dc:identifier>
<dc:title><![CDATA[Resolving chaperone-assisted protein folding on the ribosome at the peptide level]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.09.27.509658v1?rss=1">
<title>
<![CDATA[
Small-molecule Polθ inhibitors provide safe and effective tumor radiosensitization in preclinical models 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.09.27.509658v1?rss=1"
</link>
<description><![CDATA[
DNA polymerase theta (Pol{theta}) is a DNA repair enzyme critical for microhomology mediated end joining (MMEJ). Pol{theta} has limited expression in normal tissues but is frequently overexpressed in cancer cells and, therefore, represents an ideal target for tumor-specific radiosensitization. Here, we show that ART558 and ART899, two novel and specific allosteric inhibitors of the Pol{theta} DNA polymerase domain, potently radiosensitize tumor cells, particularly when combined with fractionated radiation. Importantly, normal fibroblasts are not radiosensitized by Pol{theta} inhibition. Mechanistically, we show that the radiosensitization caused by Pol{theta} inhibition is most effective in replicating cells and is due to impaired DNA damage repair. We also show that radiosensitization is still effective under hypoxia, suggesting that these inhibitors may help overcome hypoxia-induced radioresistance. In addition, we describe for the first time ART899 and characterize it as a potent and specific Pol{theta} inhibitor with improved metabolic stability. In vivo, the combination of Pol{theta} inhibition using ART899 with fractionated radiation is well tolerated and results in a significant reduction in tumor growth compared to radiation alone. These results pave the way for future clinical trials of Pol{theta} inhibitors in combination with radiotherapy.
]]></description>
<dc:creator>Rodriguez-Berriguete, G.</dc:creator>
<dc:creator>Ranzani, M.</dc:creator>
<dc:creator>Prevo, R.</dc:creator>
<dc:creator>Puliyadi, R.</dc:creator>
<dc:creator>Machado, N.</dc:creator>
<dc:creator>Bolland, H. R.</dc:creator>
<dc:creator>Millar, V.</dc:creator>
<dc:creator>Ebner, D.</dc:creator>
<dc:creator>Boursier, M.</dc:creator>
<dc:creator>Cerutti, A.</dc:creator>
<dc:creator>Cicconi, A.</dc:creator>
<dc:creator>Galbiati, A.</dc:creator>
<dc:creator>Grande, D.</dc:creator>
<dc:creator>Grinkevich, V.</dc:creator>
<dc:creator>Majithiya, J.</dc:creator>
<dc:creator>Piscitello, D.</dc:creator>
<dc:creator>Rajendra, E.</dc:creator>
<dc:creator>Stockley, M.</dc:creator>
<dc:creator>Boulton, S. J.</dc:creator>
<dc:creator>Hammond, E. M.</dc:creator>
<dc:creator>Heald, R.</dc:creator>
<dc:creator>Smith, G. C. M.</dc:creator>
<dc:creator>Robinson, H.</dc:creator>
<dc:creator>Higgins, G. S.</dc:creator>
<dc:date>2022-09-28</dc:date>
<dc:identifier>doi:10.1101/2022.09.27.509658</dc:identifier>
<dc:title><![CDATA[Small-molecule Polθ inhibitors provide safe and effective tumor radiosensitization in preclinical models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.09.27.509679v1?rss=1">
<title>
<![CDATA[
m6A-ELISA, a simple method for quantifying N6-methyladenosine from mRNA populations 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.09.27.509679v1?rss=1"
</link>
<description><![CDATA[
N6-methyladenosine (m6A) is a widely studied and abundant RNA modification. The m6A mark regulates the fate of RNAs in various ways, which in turn, drives changes in cell physiology, development, and disease pathology. Over the last decade, numerous methods have been developed to map and quantify m6A sites genomewide through deep sequencing. Alternatively, m6A levels can be quantified from a population of RNAs using techniques such as liquid chromatography-mass spectrometry or thin layer chromatography. However, many methods for quantifying m6A levels involve extensive protocols and specialized data analysis, and often only a few samples can be handled in a single experiment. Here, we developed a simple method for determining m6A levels in mRNA populations from various sources based on enzyme-linked immunosorbent-based assay (m6A-ELISA). We have optimized various steps of m6A-ELISA such as sample preparation and the background signal resulting from the primary antibody. We validated the method using mRNA populations from budding yeast and mouse embryonic stem cells. The full protocol takes less than a day, requiring only 25 ng of mRNA. The m6A-ELISA protocol is therefore quick, cost-effective, and scalable, making it a valuable tool for determining relative m6A levels in samples from various sources that could be adapted to detect other mRNA modifications.
]]></description>
<dc:creator>Ensinck, I.</dc:creator>
<dc:creator>Sideri, T.</dc:creator>
<dc:creator>Modic, M.</dc:creator>
<dc:creator>Capitanchik, C.</dc:creator>
<dc:creator>Toolan-Kerr, P.</dc:creator>
<dc:creator>van Werven, F.</dc:creator>
<dc:date>2022-09-28</dc:date>
<dc:identifier>doi:10.1101/2022.09.27.509679</dc:identifier>
<dc:title><![CDATA[m6A-ELISA, a simple method for quantifying N6-methyladenosine from mRNA populations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.09.28.509985v1?rss=1">
<title>
<![CDATA[
Treenome Browser: co-visualization of enormous phylogenies and millions of genomes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.09.28.509985v1?rss=1"
</link>
<description><![CDATA[
Treenome Browser is a web browser tool to interactively visualize millions of genomes alongside huge phylogenetic trees.

Availability and ImplementationTreenome Browser for SARS-CoV-2 can be accessed at cov2tree.org, or at taxonium.org for user-provided trees. Source code and documentation are available at github.com/theosanderson/taxonium and docs.taxonium.org/en/latest/treenome.html.

Contactalex.kramer@ucsc.edu, rucorbet@ucsc.edu
]]></description>
<dc:creator>Kramer, A. M.</dc:creator>
<dc:creator>Sanderson, T.</dc:creator>
<dc:creator>Corbett-Detig, R.</dc:creator>
<dc:date>2022-09-30</dc:date>
<dc:identifier>doi:10.1101/2022.09.28.509985</dc:identifier>
<dc:title><![CDATA[Treenome Browser: co-visualization of enormous phylogenies and millions of genomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.09.29.509946v1?rss=1">
<title>
<![CDATA[
Lineage segregation in human pre-implantation embryos is specified by YAP1 and TEAD1 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.09.29.509946v1?rss=1"
</link>
<description><![CDATA[
We know that polarity and YAP1 play a key role in trophectoderm initiation in compacted human embryos, however we know little about the TEAD family of transcription factors that become activated by YAP1 and especially if they play a role during epiblast and primitive endoderm formation. Here we show that compaction occurs heterogeneously between the 8- and 16-cell stages. While 8-cell stage blastomeres are not yet polarized, polarized outer cells and non-polarized inner cells arise in compacted 16-cell stage embryos. While trophectoderm specifiers TEAD1, YAP1 and GATA3 mostly co-localise in the nuclei of polarized outer/trophectoderm cells, they are also found in some cells of compacting embryos before polarity is established indicating that differentiation into trophectoderm cells can be initiated independently of polarity. In the inner cell mass, TEAD1 and YAP1 also distinguish GATA4 positive cells in a salt-and-pepper distribution and in the sorted primitive endoderm cells. Our detailed roadmap on polarization, compaction, position and lineage segregation events during human preimplantation development paves the road for further functional studies. Fundamental knowledge of lineage segregation events will eventually explain how and why embryos fail to develop further before or during implantation.
]]></description>
<dc:creator>Regin, M.</dc:creator>
<dc:creator>Essahib, W.</dc:creator>
<dc:creator>Demtschenko, A.</dc:creator>
<dc:creator>Dewandre, D.</dc:creator>
<dc:creator>David, L.</dc:creator>
<dc:creator>Gerri, C.</dc:creator>
<dc:creator>Niakan, K.</dc:creator>
<dc:creator>Verheyen, G.</dc:creator>
<dc:creator>Tournaye, H.</dc:creator>
<dc:creator>Sterckx, J.</dc:creator>
<dc:creator>Sermon, K.</dc:creator>
<dc:creator>Van de Velde, H.</dc:creator>
<dc:date>2022-09-30</dc:date>
<dc:identifier>doi:10.1101/2022.09.29.509946</dc:identifier>
<dc:title><![CDATA[Lineage segregation in human pre-implantation embryos is specified by YAP1 and TEAD1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.10.09.510609v1?rss=1">
<title>
<![CDATA[
ASCL1 interacts with the mSWI/SNF at distal regulatory elements to regulate neural differentiation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.10.09.510609v1?rss=1"
</link>
<description><![CDATA[
AO_SCPLOWBSTRACTC_SCPLOWPioneer transcription factors are thought to play pivotal roles in developmental processes by binding nucleosomal DNA to activate gene expression. The role of neurogenic pioneer factor ASCL1 in shaping chromatin landscape in human neurogenesis remains unclear. Here we show that ASCL1 acts as a pioneer transcription factor in a transient population of progenitors. Using an in vitro ASCL1 knockout model we show it drives progenitor differentiation by cis-regulation both as a classical pioneer factor and as a non-pioneer remodeler, where ASCL1 binds permissive chromatin to induce chromatin conformation changes. We find ASCL1 directly interacts with mammalian BAF SWI/SNF chromatin remodeling complexes, essential for neurogenesis and involved in multiple neurodevelopmental disorders. ASCL1 acts as a non-pioneer chromatin remodeler to regulate gene expression at a subset of loci, requiring mBAF SWI/SNFs ATPase activity for cis-regulation of gene expression. Our findings demonstrate that ASCL1 is a key chromatin remodeler in human neurogenesis, uncovering an alternative mechanism of remodeling function dependent on partner ATPase activity.
]]></description>
<dc:creator>Paun, O.</dc:creator>
<dc:creator>Tan, Y. X.</dc:creator>
<dc:creator>Patel, H.</dc:creator>
<dc:creator>Strohbuecker, S.</dc:creator>
<dc:creator>Ghanate, A.</dc:creator>
<dc:creator>Cobolli Gigli, C.</dc:creator>
<dc:creator>Sopena, M. L.</dc:creator>
<dc:creator>Gerontogianni, L.</dc:creator>
<dc:creator>Goldstone, R.</dc:creator>
<dc:creator>Ang, S.-L.</dc:creator>
<dc:creator>Guillemot, F.</dc:creator>
<dc:creator>Dias, C.</dc:creator>
<dc:date>2022-10-09</dc:date>
<dc:identifier>doi:10.1101/2022.10.09.510609</dc:identifier>
<dc:title><![CDATA[ASCL1 interacts with the mSWI/SNF at distal regulatory elements to regulate neural differentiation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.10.10.511318v1?rss=1">
<title>
<![CDATA[
Genome-wide RNA binding analysis of C9orf72 poly(PR) dipeptides 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.10.10.511318v1?rss=1"
</link>
<description><![CDATA[
An intronic GGGGCC repeat expansion in C9orf72 is a common genetic cause of amyotrophic lateral sclerosis and frontotemporal dementia. The repeats are transcribed in both sense and antisense directions to generate distinct dipeptide repeat proteins, of which poly(GA), poly(GR) and poly(PR) have been implicated in contributing to neurodegeneration. Poly(PR) binding to RNA may contribute to toxicity, but analysis of poly(PR)-RNA binding on a genome-wide scale has not yet been carried out. We therefore performed crosslinking and immunoprecipitation (CLIP) analysis in human cells to identify the RNA binding sites of poly(PR). We found that poly(PR) binds to nearly 600 RNAs, with the sequence GAAGA enriched at the binding sites. In vitro experiments showed that polyGAAGA RNA binds poly(PR) with higher affinity than control RNA and induces phase-separation of poly(PR) into condensates. These data indicate that poly(PR) preferentially binds to polyGAAGA-containing RNAs, which may have physiological consequences.
]]></description>
<dc:creator>Balendra, R.</dc:creator>
<dc:creator>Ruiz de los Mozos, I.</dc:creator>
<dc:creator>Glaria, I.</dc:creator>
<dc:creator>Milioto, C.</dc:creator>
<dc:creator>Odeh, H. M.</dc:creator>
<dc:creator>Wilson, K. M.</dc:creator>
<dc:creator>Ule, A. M.</dc:creator>
<dc:creator>Hallegger, M.</dc:creator>
<dc:creator>Masino, L.</dc:creator>
<dc:creator>Martin, S.</dc:creator>
<dc:creator>Patani, R.</dc:creator>
<dc:creator>Shorter, J.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Isaacs, A.</dc:creator>
<dc:date>2022-10-11</dc:date>
<dc:identifier>doi:10.1101/2022.10.10.511318</dc:identifier>
<dc:title><![CDATA[Genome-wide RNA binding analysis of C9orf72 poly(PR) dipeptides]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.10.12.511600v1?rss=1">
<title>
<![CDATA[
Restoration of functional PAX6 in aniridia patient iPSC-derived ocular tissue models using repurposed nonsense suppression drugs 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.10.12.511600v1?rss=1"
</link>
<description><![CDATA[
Aniridia is a rare, pan-ocular disease causing severe sight loss, with only symptomatic intervention offered to patients. Approximately 40% of aniridia patients present with heterozygous nonsense variants in PAX6, resulting in haploinsufficiency. Translational readthrough inducing compounds (TRIDs) have the ability to weaken the recognition of in-frame premature stop codons (PTCs), permitting full-length protein to be translated. We have established induced pluripotent stem cell (iPSC)-derived 3D optic cups and 2D limbal epithelial stem cell (LESC) models from an aniridia patient with a prevalent PAX6 nonsense mutation. Both in vitro models show reduced PAX6 protein levels, mimicking the disease. Repurposed TRIDs amlexanox and 2,6-diaminopurine (DAP), and positive control compounds ataluren and G418 were tested for their efficiency. Amlexanox was identified as the most promising TRID, increasing full-length PAX6 levels in both models, and rescuing the disease phenotype through normalization of VSX2 and cell proliferation in the optic cups and reduction of ABCG2 protein and SOX10 expression in LESC. This study highlights the significance of patient iPSC-derived cells as a new model system for aniridia and proposes amlexanox as a new putative treatment for nonsense-mediated aniridia.
]]></description>
<dc:creator>Lima Cunha, D.</dc:creator>
<dc:creator>Eintracht, J.</dc:creator>
<dc:creator>Harding, P.</dc:creator>
<dc:creator>Zhou, J. H.</dc:creator>
<dc:creator>Moosajee, M.</dc:creator>
<dc:date>2022-10-12</dc:date>
<dc:identifier>doi:10.1101/2022.10.12.511600</dc:identifier>
<dc:title><![CDATA[Restoration of functional PAX6 in aniridia patient iPSC-derived ocular tissue models using repurposed nonsense suppression drugs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.10.17.512486v1?rss=1">
<title>
<![CDATA[
Rates of global cellular translation and transcription during cell growth and the cell cycle in fission yeast. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.10.17.512486v1?rss=1"
</link>
<description><![CDATA[
Proliferating eukaryotic cells grow and undergo cycles of cell division. Growth is continuous whilst the cell cycle consists of discrete events. How the production of biomass is controlled as cells increase in size and proceed through the cell cycle is important for understanding the regulation of global cellular growth. This has been studied for decades but has not yielded consistent results. Previous studies investigating how cell size, the amount of DNA, and cell cycle events affect the global cellular production of proteins and RNA molecules have led to highly conflicting results, probably due to perturbations induced by the synchronisation methods used. To avoid these perturbations, we have developed a system to assay unperturbed exponentially growing populations of fission yeast cells. We generated thousands of single-cell measurements of cell size, of cell cycle stage, and of the levels of global cellular translation and transcription. This has allowed us to determine how cellular changes arising from progression through the cell cycle and cells growing in size affect global cellular translation and transcription. We show that translation scales with size, and additionally increases at late S-phase/early G2, then increases early in mitosis and decreases later in mitosis, suggesting that cell cycle controls are operative over global cellular translation. Transcription increases with both size and the amount of DNA, suggesting that the level of transcription of a cell may be the result of a dynamic equilibrium between the number of RNA polymerases associating and disassociating from DNA.
]]></description>
<dc:creator>Basier, C.</dc:creator>
<dc:creator>Nurse, P.</dc:creator>
<dc:date>2022-10-17</dc:date>
<dc:identifier>doi:10.1101/2022.10.17.512486</dc:identifier>
<dc:title><![CDATA[Rates of global cellular translation and transcription during cell growth and the cell cycle in fission yeast.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.10.13.511885v1?rss=1">
<title>
<![CDATA[
Refphase: Multi-sample reference phasing reveals haplotype-specific copy number heterogeneity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.10.13.511885v1?rss=1"
</link>
<description><![CDATA[
Most computational methods that infer somatic copy number alterations (SCNAs) from bulk sequencing of DNA analyse tumour samples individually. However, the sequencing of multiple tumour samples from a patients disease is an increasingly common practice. We introduce Refphase, an algorithm that leverages this multi-sampling approach to infer haplotype-specific copy numbers through multi-sample reference phasing. We demonstrate Refphases ability to infer haplotype-specific SCNAs and characterise their intra-tumour heterogeneity, to uncover previously undetected allelic imbalance in low purity samples, and to identify parallel evolution in the context of whole genome doubling in a pan-cancer cohort of 336 samples from 99 tumours.
]]></description>
<dc:creator>Watkins, T. B.</dc:creator>
<dc:creator>Colliver, E. C.</dc:creator>
<dc:creator>Huska, M. R.</dc:creator>
<dc:creator>Kaufmann, T. L.</dc:creator>
<dc:creator>Lim, E. L.</dc:creator>
<dc:creator>Haase, K.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>McGranahan, N.</dc:creator>
<dc:creator>Schwarz, R. F.</dc:creator>
<dc:date>2022-10-17</dc:date>
<dc:identifier>doi:10.1101/2022.10.13.511885</dc:identifier>
<dc:title><![CDATA[Refphase: Multi-sample reference phasing reveals haplotype-specific copy number heterogeneity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.10.19.512927v1?rss=1">
<title>
<![CDATA[
Global landscape of the host response to SARS-CoV-2 variants reveals viral evolutionary trajectories 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.10.19.512927v1?rss=1"
</link>
<description><![CDATA[
A series of SARS-CoV-2 variants of concern (VOCs) have evolved in humans during the COVID-19 pandemic--Alpha, Beta, Gamma, Delta, and Omicron. Here, we used global proteomic and genomic analyses during infection to understand the molecular responses driving VOC evolution. We discovered VOC-specific differences in viral RNA and protein expression levels, including for N, Orf6, and Orf9b, and pinpointed several viral mutations responsible. An analysis of the host response to VOC infection and comprehensive interrogation of altered virus-host protein-protein interactions revealed conserved and divergent regulation of biological pathways. For example, regulation of host translation was highly conserved, consistent with suppression of VOC replication in mice using the translation inhibitor plitidepsin. Conversely, modulation of the host inflammatory response was most divergent, where we found Alpha and Beta, but not Omicron BA.1, antagonized interferon stimulated genes (ISGs), a phenotype that correlated with differing levels of Orf6. Additionally, Delta more strongly upregulated proinflammatory genes compared to other VOCs. Systematic comparison of Omicron subvariants revealed BA.5 to have evolved enhanced ISG and proinflammatory gene suppression that similarly correlated with Orf6 expression, effects not seen in BA.4 due to a mutation that disrupts the Orf6-nuclear pore interaction. Our findings describe how VOCs have evolved to fine-tune viral protein expression and protein-protein interactions to evade both innate and adaptive immune responses, offering a likely explanation for increased transmission in humans.

One sentence summarySystematic proteomic and genomic analyses of SARS-CoV-2 variants of concern reveal how variant-specific mutations alter viral gene expression, virus-host protein complexes, and the host response to infection with applications to therapy and future pandemic preparedness.
]]></description>
<dc:creator>Bouhaddou, M.</dc:creator>
<dc:creator>Reuschl, A.-K.</dc:creator>
<dc:creator>Polacco, B. J.</dc:creator>
<dc:creator>Thorne, L. G.</dc:creator>
<dc:creator>Ummadi, M. R.</dc:creator>
<dc:creator>Ye, C.</dc:creator>
<dc:creator>Ramirez, R. R.</dc:creator>
<dc:creator>Pelin, A.</dc:creator>
<dc:creator>Batra, J.</dc:creator>
<dc:creator>Jang, G. M.</dc:creator>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Moen, J. M.</dc:creator>
<dc:creator>Richards, A. L.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Harjai, B.</dc:creator>
<dc:creator>Stevenson, E.</dc:creator>
<dc:creator>Rojc, A.</dc:creator>
<dc:creator>Ragazzini, R.</dc:creator>
<dc:creator>Whelan, M. V. X.</dc:creator>
<dc:creator>Furnon, W.</dc:creator>
<dc:creator>De Lorenzo, G.</dc:creator>
<dc:creator>Cowton, V.</dc:creator>
<dc:creator>Syed, A. M.</dc:creator>
<dc:creator>Ciling, A.</dc:creator>
<dc:creator>Deutsch, N.</dc:creator>
<dc:creator>Pirak, D.</dc:creator>
<dc:creator>Dowgier, G.</dc:creator>
<dc:creator>Mesner, D.</dc:creator>
<dc:creator>Turner, J. L.</dc:creator>
<dc:creator>McGovern, B. L.</dc:creator>
<dc:creator>Rodriguez, M. L.</dc:creator>
<dc:creator>Leiva-Rebollo, R.</dc:creator>
<dc:creator>Dunham, A. S.</dc:creator>
<dc:creator>Zhong, X.</dc:creator>
<dc:creator>Eckhardt, M.</dc:creator>
<dc:creator>Fossati, A.</dc:creator>
<dc:creator>Liotta, N.</dc:creator>
<dc:creator>Kehrer, T.</dc:creator>
<dc:creator>Cupic, A.</dc:creator>
<dc:creator>Rutkowska, M.</dc:creator>
<dc:creator>Mena, N.</dc:creator>
<dc:creator>Aslam, S.</dc:creator>
<dc:creator>Hoffert, A.</dc:creator>
<dc:creator>Foussard, H.</dc:creator>
<dc:creator>Pham, J.</dc:creator>
<dc:creator>Ly</dc:creator>
<dc:date>2022-10-21</dc:date>
<dc:identifier>doi:10.1101/2022.10.19.512927</dc:identifier>
<dc:title><![CDATA[Global landscape of the host response to SARS-CoV-2 variants reveals viral evolutionary trajectories]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.10.21.513169v1?rss=1">
<title>
<![CDATA[
Pseudogenes limit the identification of novel common transcripts generated by their parent genes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.10.21.513169v1?rss=1"
</link>
<description><![CDATA[
The human genome contains numerous duplicated regions, such as parent-pseudogene pairs, causing sequencing reads to align equally well to either gene. The extent to which this ambiguity complicates transcriptomic analyses is currently unknown. This is concerning as many parent genes have been linked to disease, including GBA1, causally linked to both Parkinsons and Gaucher disease. We find that most of the short sequencing reads that map to GBA1, also map to its pseudogene, GBAP1. Using long-read RNA-sequencing in human brain, where all reads mapped uniquely, we demonstrate significant differences in expression compared to short-read data. We identify novel transcripts from both GBA1 and GBAP1, including protein-coding transcripts that are translated in vitro and detected in proteomic data, but that lack GCase activity. By combining long-read with single-nuclear RNA-sequencing to analyse brain-relevant cell types we demonstrate that transcript expression varies by brain region with cell-type-selectivity. Taken together, these results suggest a non-lysosomal function for both GBA1 and GBAP1 in brain. Finally, we demonstrate that inaccuracies in annotation are widespread among parent genes, with implications for many human diseases.
]]></description>
<dc:creator>Gustavsson, E. K.</dc:creator>
<dc:creator>Sethi, S.</dc:creator>
<dc:creator>Gao, Y.</dc:creator>
<dc:creator>Brenton, J.</dc:creator>
<dc:creator>Garcia Ruiz, S.</dc:creator>
<dc:creator>Zhang, D.</dc:creator>
<dc:creator>Garza, R.</dc:creator>
<dc:creator>Reynolds, R. H.</dc:creator>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Grant-Peters, M.</dc:creator>
<dc:creator>MacPherson, H.</dc:creator>
<dc:creator>Montgomery, K.</dc:creator>
<dc:creator>Dore, R.</dc:creator>
<dc:creator>Wernick, A. I.</dc:creator>
<dc:creator>Arber, C.</dc:creator>
<dc:creator>Wray, S.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Esselborn, J.</dc:creator>
<dc:creator>Blauwendraat, C.</dc:creator>
<dc:creator>Douse, C. H.</dc:creator>
<dc:creator>Adami, A.</dc:creator>
<dc:creator>Atacho, D. A. M.</dc:creator>
<dc:creator>Kouli, A.</dc:creator>
<dc:creator>Quaegebeur, A.</dc:creator>
<dc:creator>Barker, R. A.</dc:creator>
<dc:creator>Englund, E.</dc:creator>
<dc:creator>Platt, F. M.</dc:creator>
<dc:creator>Jakobsson, J.</dc:creator>
<dc:creator>Wood, N. W.</dc:creator>
<dc:creator>Houlden, H.</dc:creator>
<dc:creator>Saini, H.</dc:creator>
<dc:creator>Bento, C. F.</dc:creator>
<dc:creator>Hardy, J.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:date>2022-10-21</dc:date>
<dc:identifier>doi:10.1101/2022.10.21.513169</dc:identifier>
<dc:title><![CDATA[Pseudogenes limit the identification of novel common transcripts generated by their parent genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.10.21.513232v1?rss=1">
<title>
<![CDATA[
Blood and site of disease inflammatory profiles differ in HIV-1-infected pericardial tuberculosis patients 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.10.21.513232v1?rss=1"
</link>
<description><![CDATA[
ObjectivesTo better understand the pathogenesis of pericardial tuberculosis (PCTB), we sought to characterize the systemic inflammatory profile in HIV-1-infected participants with latent TB infection (LTBI), pulmonary TB (PTB) and PCTB.

MethodsUsing Luminex, we measured 39 analytes in pericardial fluid (PCF) and paired plasma from 18 PCTB participants, and plasma from 16 LTBI and 20 PTB. Follow-up plasma samples were also obtained from PTB and PCTB participants. HLA-DR expression on Mtb-specific CD4 T cells was measured in baseline samples using flow cytometry.

ResultsAssessment of the overall systemic inflammatory profile by principal component analysis showed that the inflammatory profile of active TB participants was distinct from the LTBI group, while PTB patients could not be distinguished from those with PCTB. In the LTBI group, 12 analytes showed a positive association with plasma HIV-1 viral load, and most of these associations were lost in the diseased groups. When comparing the inflammatory profile between PCF and paired blood, we found that the concentrations of most analytes (24/39) were elevated at site of disease. However, the inflammatory profile in PCF partially mirrored inflammatory events in the blood. After TB treatment completion, the overall plasma inflammatory profile reverted to those observed in the LTBI group. Lastly, HLA-DR expression showed the best performance for TB diagnosis compared to previously described biosignatures built from soluble markers.

ConclusionOur results describe the inflammatory profile associated with PTB and PCTB and emphasize the potential role of HLA-DR as a promising biomarker for TB diagnosis.
]]></description>
<dc:creator>Mutavhatsindi, H.</dc:creator>
<dc:creator>Du Bruyn, E.</dc:creator>
<dc:creator>Ruzive, S.</dc:creator>
<dc:creator>Howlett, P.</dc:creator>
<dc:creator>Sher, A.</dc:creator>
<dc:creator>Mayer-Barber, K. D.</dc:creator>
<dc:creator>Barber, D. L.</dc:creator>
<dc:creator>Ntsekhe, M.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Riou, C.</dc:creator>
<dc:date>2022-10-24</dc:date>
<dc:identifier>doi:10.1101/2022.10.21.513232</dc:identifier>
<dc:title><![CDATA[Blood and site of disease inflammatory profiles differ in HIV-1-infected pericardial tuberculosis patients]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.10.24.513556v1?rss=1">
<title>
<![CDATA[
A palmitate-rich metastatic niche enables metastasis growth via p65 acetylation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.10.24.513556v1?rss=1"
</link>
<description><![CDATA[
Cancer cells outgrowing in distant organs of metastasis rewire their metabolism to fuel on the available nutrients. While this is often considered an adaptive pressure limiting metastasis formation, some nutrients available at the metastatic site naturally or through changes in organ physiology may inherently promote metastatic growth. We find that the lung, a frequent site of metastasis, is a lipid-rich environment. Moreover, we observe that pathological conditions such as pre-metastatic niche formation and obesity further increase the availability of the fatty acid palmitate in the lung. We find that targeting palmitate processing inhibits spheroid growth in vitro and metastasis formation in lean and obese mice. Mechanistically, we discover that breast cancer cells use palmitate to synthesize acetyl-CoA in a carnitine palmitoyltransferase 1a (CPT1a)-dependent manner. Lysine acetyltransferase 2a (KAT2a), whose expression is promoted by palmitate availability, relies on the available acetyl-CoA to acetylate the NF-{kappa}B subunit p65. This favors nuclear location of p65 and activates a pro-metastatic transcriptional program. Accordingly, deletion of KAT2a phenocopies CPT1a silencing in vitro as well as in vivo and patients with breast cancer show co-expression of both proteins in metastases across palmitate-rich metastatic sites. In conclusion, we find that palmitate-rich environments foster metastasis growth by increasing p65 acetylation resulting in elevated NF-{kappa}B signaling.
]]></description>
<dc:creator>Altea-Manzano, P.</dc:creator>
<dc:creator>Doglioni, G.</dc:creator>
<dc:creator>Cuadros, A. M.</dc:creator>
<dc:creator>Nolan, E.</dc:creator>
<dc:creator>Fernandez-Garcia, J.</dc:creator>
<dc:creator>Wu, Q.</dc:creator>
<dc:creator>Cidre-Aranaz, F.</dc:creator>
<dc:creator>Montagne, A.</dc:creator>
<dc:creator>Planque, M.</dc:creator>
<dc:creator>Marin-Bejar, O.</dc:creator>
<dc:creator>Van Elsen, J.</dc:creator>
<dc:creator>Vermeire, I.</dc:creator>
<dc:creator>Broekaert, D.</dc:creator>
<dc:creator>Riera-Domingo, C.</dc:creator>
<dc:creator>Richard, F.</dc:creator>
<dc:creator>Geukens, T.</dc:creator>
<dc:creator>De Schepper, M.</dc:creator>
<dc:creator>Leduc, S.</dc:creator>
<dc:creator>Hatse, S.</dc:creator>
<dc:creator>Lambrechts, Y.</dc:creator>
<dc:creator>Kay, E.</dc:creator>
<dc:creator>Lilla, S.</dc:creator>
<dc:creator>Demeyer, S.</dc:creator>
<dc:creator>Geldhof, V.</dc:creator>
<dc:creator>Boeckx, B.</dc:creator>
<dc:creator>Alekseenko, A.</dc:creator>
<dc:creator>de la Calle Arregui, C.</dc:creator>
<dc:creator>Floris, G.</dc:creator>
<dc:creator>Marine, J.-C.</dc:creator>
<dc:creator>Lambrechts, D.</dc:creator>
<dc:creator>Pelechano, V.</dc:creator>
<dc:creator>Mazzone, M.</dc:creator>
<dc:creator>Zanivan, S.</dc:creator>
<dc:creator>Cools, J.</dc:creator>
<dc:creator>Wildiers, H.</dc:creator>
<dc:creator>Baud, V.</dc:creator>
<dc:creator>Gruenewald, T. G. P.</dc:creator>
<dc:creator>Desmedt, C.</dc:creator>
<dc:creator>Malanchi, I.</dc:creator>
<dc:creator>Fendt, S.-M.</dc:creator>
<dc:date>2022-10-24</dc:date>
<dc:identifier>doi:10.1101/2022.10.24.513556</dc:identifier>
<dc:title><![CDATA[A palmitate-rich metastatic niche enables metastasis growth via p65 acetylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.10.24.513438v1?rss=1">
<title>
<![CDATA[
The identification of a gene expression signature of primordial follicle activation in mouse pregranulosa cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.10.24.513438v1?rss=1"
</link>
<description><![CDATA[
Primordial follicles are quiescent ovarian structures comprised of a single oocyte surrounded by a layer of somatic supporting pregranulosa cells. Primordial follicle activation is the first step towards oocyte maturation and, ultimately, ovulation. As the number of quiescent primordial follicles is finite, their rate of activation is a critical parameter of the duration of the female reproductive lifespan. Activation status is established by the presence of cuboidal and proliferative granulosa cells in primary follicles, rather than squamous and quiescent pregranulosa cells in primordial follicles. Here, using a continuous Entropy Sort Feature Weighting approach on single-cell RNA sequencing data, we identify a distinct transcriptomic signature of activating pregranulosa cells in neonatal wildtype mice. This signature contains several genes previously linked with mature granulosa cells as well several novel candidates: Slc18a2, Tnni3, Fam13a and Myo1e. We confirm expression of Slc18a2 and TNNI3 in the granulosa cells of activating follicles in embryonic, neonatal and adult mouse ovaries. Perturbation of cell cycle inhibitor p27kip1 in Cdkn1b-/- mice results in complete activation of all primordial follicles during this neonatal period. Contrary to previous reports on this established mouse model, we find substantial transcriptomic changes in embryonic Cdkn1b-/- ovaries. Upon loss of cell-cycle inhibition, we find increased expression of our signature of pregranulosa cell activation, particularly that of cardiac troponin I (Tnni3). We conclude that pregranulosa cells engage a distinct transcriptional programme prior to cell-cycle dependent primordial follicle activation.
]]></description>
<dc:creator>Frost, E. R.</dc:creator>
<dc:creator>Taylor, G.</dc:creator>
<dc:creator>Boeing, S.</dc:creator>
<dc:creator>Galichet, C.</dc:creator>
<dc:creator>Baker, M. A.</dc:creator>
<dc:creator>Sutherland, J. M.</dc:creator>
<dc:creator>Lovell-Badge, R.</dc:creator>
<dc:date>2022-10-24</dc:date>
<dc:identifier>doi:10.1101/2022.10.24.513438</dc:identifier>
<dc:title><![CDATA[The identification of a gene expression signature of primordial follicle activation in mouse pregranulosa cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.11.02.514842v1?rss=1">
<title>
<![CDATA[
Intracellular calcium elevations drive the nucleation of FIP200- and ATG13-containing pre-autophagosomal structures that become omegasomes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.02.514842v1?rss=1"
</link>
<description><![CDATA[
Ca2+ modulates autophagy at multiple steps including the induction and maturation of autophagosomes, but the magnitude and spatiotemporal properties of this calcium signal and its ultimate effect on the autophagic machinery are unclear. Focusing on the induction step leading to omegasome formation, we report that low but sustained elevations in cytosolic calcium levels induce omegasome formation but treatments that only transiently elevate calcium do not. The calcium-induced structures are early intermediates that mature into omegasomes but do not constitute full autophagosomes because they are partially devoid of late autophagy proteins ATG16 and LC3. In addition to omegasomes, all four components of the ULK complex (ULK1, FIP200, ATG13, ATG101) respond to calcium modulation: they translocate to early autophagy puncta in complete medium upon calcium elevation, and are inhibited from translocation during starvation by calcium chelation with BAPTA-2 AM. The principal early step affected by calcium lies downstream of mTORC1 inactivation and upstream of VPS34 activation, coinciding biochemically with phosphorylation of ATG13 at serine 318, which is known to require ULK1 activity. However, although the calcium-mediated step requires ATG9, FIP200 and ATG13, it does not require ULK1/2, suggesting that calcium does not directly regulate ULK1 activity but rather it regulates the mechanism by which the ULK complex components ATG13 and FIP200, together with ATG9, nucleate pre-autophagosomal precursors. This calcium-induced nucleation is sufficient to drive autophagy induction up to the omegasome step, but not beyond it.
]]></description>
<dc:creator>Smith, M.</dc:creator>
<dc:creator>Schoenfelder, P.</dc:creator>
<dc:creator>Manifava, M.</dc:creator>
<dc:creator>Polson, H.</dc:creator>
<dc:creator>Tooze, S.</dc:creator>
<dc:creator>Roderick, L.</dc:creator>
<dc:creator>Ktistakis, N.</dc:creator>
<dc:date>2022-11-02</dc:date>
<dc:identifier>doi:10.1101/2022.11.02.514842</dc:identifier>
<dc:title><![CDATA[Intracellular calcium elevations drive the nucleation of FIP200- and ATG13-containing pre-autophagosomal structures that become omegasomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.10.27.514062v1?rss=1">
<title>
<![CDATA[
HNRNPH1 regulates the neuroprotective cold-shock protein RBM3 expression through poison exon exclusion 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.10.27.514062v1?rss=1"
</link>
<description><![CDATA[
Enhanced expression of the cold-shock protein RNA binding motif 3 (RBM3) is highly neuroprotective both in vitro and in vivo. Whilst upstream signalling pathways leading to RBM3 expression have been described, the precise molecular mechanism of RBM3 induction during cooling remains elusive. To identify temperature-dependent modulators of RBM3, we performed a genome-wide CRISPR-Cas9 knockout screen using RBM3-reporter human iPSC-derived neurons. We found that RBM3 mRNA and protein levels are robustly regulated by several splicing factors, with heterogeneous nuclear ribonucleoprotein H1 (HNRNPH1) being the strongest positive regulator. Splicing analysis revealed that moderate hypothermia significantly represses the inclusion of a poison exon, which, when retained, targets the mRNA for nonsense-mediated decay. Importantly, we show that HNRNPH1 mediates this cold-dependent exon skipping via its interaction with a G-rich motif within the poison exon. Our study provides novel mechanistic insights into the regulation of RBM3 and provides further targets for neuroprotective therapeutic strategies.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/514062v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Lin, J. Q.</dc:creator>
<dc:creator>Khuperkar, D.</dc:creator>
<dc:creator>Pavlou, S.</dc:creator>
<dc:creator>Makarchuk, S.</dc:creator>
<dc:creator>Patikas, N.</dc:creator>
<dc:creator>Lee, F. C. Y.</dc:creator>
<dc:creator>Kang, J.</dc:creator>
<dc:creator>Field, S. F.</dc:creator>
<dc:creator>Zbiegly, J. M.</dc:creator>
<dc:creator>Freeman, J. L.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Metzakopian, E.</dc:creator>
<dc:creator>Ruepp, M.-D.</dc:creator>
<dc:creator>Mallucci, G. R.</dc:creator>
<dc:date>2022-10-27</dc:date>
<dc:identifier>doi:10.1101/2022.10.27.514062</dc:identifier>
<dc:title><![CDATA[HNRNPH1 regulates the neuroprotective cold-shock protein RBM3 expression through poison exon exclusion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.10.28.514238v1?rss=1">
<title>
<![CDATA[
Protein aggregation and calcium dysregulation are the earliest hallmarks of synucleinopathy in human midbrain dopaminergic neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.10.28.514238v1?rss=1"
</link>
<description><![CDATA[
Mutations in the SNCA gene cause autosomal dominant Parkinsons disease (PD), with progressive loss of dopaminergic neurons in the substantia nigra, and accumulation of aggregates of -synuclein. However, the sequence of molecular events that proceed from the SNCA mutation during development, to its end stage pathology is unknown. Utilising human induced pluripotent stem cells (hiPSCs) with SNCA mutations, we resolved the temporal sequence of pathophysiological events that occur during neuronal differentiation in order to discover the early, and likely causative, events in synucleinopathies. We adapted a small molecule-based protocol that generates highly enriched midbrain dopaminergic (mDA) neurons (>80%). We characterised their molecular identity using single-cell RNA sequencing and their functional identity through the synthesis and secretion of dopamine, the ability to generate action potentials, and form functional synapses and networks. RNA velocity analyses confirmed the developmental transcriptomic trajectory of midbrain neural precursors into mDA neurons using our approach, and identified key driver genes in mDA neuronal development. To characterise the synucleinopathy, we adopted super-resolution methods to determine the number, size and structure of aggregates in SNCA-mutant mDA neurons. At one week of differentiation, prior to maturation to mDA neurons of molecular and functional identity, we demonstrate the formation of small aggregates; specifically, {beta}-sheet rich oligomeric aggregates, in SNCA-mutant midbrain immature neurons. The aggregation progresses over time to accumulate phosphorylated aggregates, and later fibrillar aggregates. When the midbrain neurons were functional, we observed evidence of impaired physiological calcium signalling, with raised basal calcium, and impairments in cytosolic and mitochondrial calcium efflux. Once midbrain identity fully developed, SNCA-mutant neurons exhibited bioenergetic impairments, mitochondrial dysfunction and oxidative stress. During the maturation of mDA neurons, upregulation of mitophagy and autophagy occured, and ultimately these multiple cellular stresses lead to an increase in cell death by six weeks post-differentiation. Our differentiation paradigm generates an efficient model for studying disease mechanisms in PD, and highlights that protein misfolding to generate intraneuronal oligomers is one of the earliest critical events driving disease in human neurons, rather than a late-stage hallmark of the disease.
]]></description>
<dc:creator>Virdi, G. S.</dc:creator>
<dc:creator>Choi, M. L.</dc:creator>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>Yao, Z.</dc:creator>
<dc:creator>Athauda, D.</dc:creator>
<dc:creator>Strohbuecker, S.</dc:creator>
<dc:creator>Wernick, A. I.</dc:creator>
<dc:creator>Alrashidi, H.</dc:creator>
<dc:creator>Melandri, D.</dc:creator>
<dc:creator>Perez-lloret, J.</dc:creator>
<dc:creator>Stroh, P. R.</dc:creator>
<dc:creator>Sylantyev, S.</dc:creator>
<dc:creator>Eaton, S.</dc:creator>
<dc:creator>Heales, S.</dc:creator>
<dc:creator>Kunath, T.</dc:creator>
<dc:creator>Horrocks, M. H.</dc:creator>
<dc:creator>Abramov, A. Y.</dc:creator>
<dc:creator>Patani, R.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:date>2022-10-28</dc:date>
<dc:identifier>doi:10.1101/2022.10.28.514238</dc:identifier>
<dc:title><![CDATA[Protein aggregation and calcium dysregulation are the earliest hallmarks of synucleinopathy in human midbrain dopaminergic neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.11.03.515001v1?rss=1">
<title>
<![CDATA[
An improved Erk biosensor reveals oscillatory Erk dynamics driven by mitotic erasure during early development 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.03.515001v1?rss=1"
</link>
<description><![CDATA[
Erk signaling dynamics elicit distinct cellular responses in a variety of contexts. The early zebrafish embryo is an ideal model to explore the role of Erk signaling dynamics in vivo, as a gradient of activated diphosphorylated Erk (P-Erk) is induced by Fgf signaling at the blastula embryonic margin. Here we describe an improved Erk-specific biosensor which we term modified Erk Kinase Translocation Reporter (modErk-KTR). We demonstrate the utility of this biosensor in vitro and in developing zebrafish and Drosophila embryos. Moreover, we show that Fgf/Erk signaling is dynamic and coupled to tissue growth during both early zebrafish and Drosophila development. Signaling is rapidly extinguished just prior to mitosis, which we refer to as mitotic erasure, inducing periods of inactivity, thus providing a source of heterogeneity in an asynchronously dividing tissue. Our modified reporter and transgenic lines represent an important resource for interrogating the role of Erk signaling dynamics in vivo.
]]></description>
<dc:creator>Wilcockson, S. G.</dc:creator>
<dc:creator>Guglielmi, L.</dc:creator>
<dc:creator>Araguas Rodriguez, P.</dc:creator>
<dc:creator>Amoyel, M.</dc:creator>
<dc:creator>Hill, C. S.</dc:creator>
<dc:date>2022-11-04</dc:date>
<dc:identifier>doi:10.1101/2022.11.03.515001</dc:identifier>
<dc:title><![CDATA[An improved Erk biosensor reveals oscillatory Erk dynamics driven by mitotic erasure during early development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.11.04.515216v1?rss=1">
<title>
<![CDATA[
Subcytoplasmic location of translation controls protein output 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.04.515216v1?rss=1"
</link>
<description><![CDATA[
The cytoplasm is highly compartmentalized, but the extent and consequences of subcytopIasmic mRNA localization in non-polarized cells are largely unknown. We determined mRNA enrichment in TIS granules (TGs) and the rough endopIasmic reticuIum (ER) through particle sorting and isolated cytosolic mRNAs by digitonin extraction. When focusing on non-membrane protein-encoding mRNAs, we observed that 52% have a biased transcript distribution across these compartments. Compartment enrichment is determined by a combinatorial code based on mRNA length, exon length, and 3'UTR-bound RNA-binding proteins. Compartment-biased mRNAs differ in the functional classes of their encoded proteins: TG-enriched mRNAs encode low-abundance proteins with strong enrichment of transcription factors, whereas ER-enriched mRNAs encode large and highly expressed proteins. Compartment localization is an important determinant of mRNA and protein abundance, which is supported by reporter experiments showing that redirecting cytosolic mRNAs to the ER increases their protein expression. In summary, the cytoplasm is functionally compartmentaIized by local translation environments.
]]></description>
<dc:creator>Horste, E. L.</dc:creator>
<dc:creator>Zhen, G.</dc:creator>
<dc:creator>Fansler, M. M.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Lee, F. C. Y.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Mayr, C.</dc:creator>
<dc:date>2022-11-04</dc:date>
<dc:identifier>doi:10.1101/2022.11.04.515216</dc:identifier>
<dc:title><![CDATA[Subcytoplasmic location of translation controls protein output]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.11.07.515440v1?rss=1">
<title>
<![CDATA[
Pituitary stem cells differ according to their position in the gland and maintenance of their progeny depends on physiological context 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.07.515440v1?rss=1"
</link>
<description><![CDATA[
Stem cell (SC) differentiation and maintenance of resultant progeny underlie cell-turnover in many organs, but it is difficult to pinpoint the contribution of either process. In the pituitary, a central regulator of endocrine axes, adult SCs undergo activation following target organ ablation, providing a well-characterized paradigm to study an adaptative response in a multi-organ system. Here we used single cell technologies to characterize SC heterogeneity and mobilization together with lineage tracing. We show that SC differentiation occurs more frequently than thought previously. In adaptative conditions, differentiation increases and is more diverse than demonstrated by the lineage tracing experiments. Detailed examination of SC progeny suggests that maintenance of selected nascent cells underlies SC output, highlighting a trophic role for the microenvironment. Analyses of cell trajectories further predict pathways and potential new regulators. Our model provides a valuable system to study the influence of evolving states on the mechanisms of SC mobilization.

Teaser: Pituitary stem cells are diverse and differentiate more than thought but only selected progeny persist according to need.
]]></description>
<dc:creator>Rizzoti, K.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Sheridan, D.</dc:creator>
<dc:creator>Lovell-Badge, R.</dc:creator>
<dc:date>2022-11-07</dc:date>
<dc:identifier>doi:10.1101/2022.11.07.515440</dc:identifier>
<dc:title><![CDATA[Pituitary stem cells differ according to their position in the gland and maintenance of their progeny depends on physiological context]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.11.03.515052v1?rss=1">
<title>
<![CDATA[
Aryl hydrocarbon receptor utilises cellular zinc signals to maintain the gut epithelial barrier 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.03.515052v1?rss=1"
</link>
<description><![CDATA[
Both zinc and plant-derived ligands of the aryl hydrocarbon receptor (AHR) are dietary components which regulate intestinal epithelial barrier function and protect against Inflammatory Bowel Disease (IBD)1,2. Here, we explore whether zinc and AHR pathway are linked using a mouse IBD model with follow-on studies on human and mouse ileum organoids. Our data demonstrate that AHR regulates cellular zinc uptake, and that zinc is an integral part of AHR signalling processes. We show that dietary supplementation in mice with the plant-derived AHR ligand precursor, indole-3-carbinol (I3C), offers a high level of protection against dextran sulfate sodium induced IBD while protection fails in mice with AHR deleted in the intestinal epithelium. AHR agonist treatment is also ineffective in mice with a nutritional zinc deficiency. Experiments in the human Caco-2 cell line and ileum organoids showed that AHR activation increases total cellular zinc and cytosolic free Zn2+ concentrations through transcriptional upregulation of several SLC39 zinc importers. As a consequence, genes for tight junction (TJ) proteins were upregulated in a zinc-dependent manner involving zinc inhibition of signalling to NF-{kappa}B and attenuated degradation of TJ proteins through zinc inhibition of calpain activity. Thus, our data indicate that AHR activation by plant-derived dietary ligands improves gut barrier function via zinc-dependent cellular pathways, suggesting that combined dietary supplementation with AHR ligands and zinc might be effective in preventing and treating inflammatory gut disorders.
]]></description>
<dc:creator>Hu, X.</dc:creator>
<dc:creator>Xiao, W.</dc:creator>
<dc:creator>Lei, Y.</dc:creator>
<dc:creator>Green, A. J.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Maradana, M. R.</dc:creator>
<dc:creator>Gao, Y.</dc:creator>
<dc:creator>Xie, X.</dc:creator>
<dc:creator>Wang, R.</dc:creator>
<dc:creator>Chennell, G.</dc:creator>
<dc:creator>Basson, M. A.</dc:creator>
<dc:creator>Kille, P.</dc:creator>
<dc:creator>Maret, W.</dc:creator>
<dc:creator>Bewick, G.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Hogstrand, C.</dc:creator>
<dc:date>2022-11-04</dc:date>
<dc:identifier>doi:10.1101/2022.11.03.515052</dc:identifier>
<dc:title><![CDATA[Aryl hydrocarbon receptor utilises cellular zinc signals to maintain the gut epithelial barrier]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.11.13.516300v1?rss=1">
<title>
<![CDATA[
Structural basis for ubiquitylation by HOIL-1 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.13.516300v1?rss=1"
</link>
<description><![CDATA[
The linear ubiquitin chain assembly complex (LUBAC) synthesises linear Ub chains which constitute a binding and activation platform for components of the TNF signalling pathway. One of the components of LUBAC is the ubiquitin ligase HOIL-1 which has been shown to generate oxyester linkages on several proteins and on linear polysaccharides. Here we describe the crystal structure of a C-terminal tandem domain construct of HOIL-1 comprising the IBR and RING2 domains. The structure adopts an auto-inhibited conformation in which the catalytic cysteine of the RING2 domain is shielded by the adjacent IBR domain. Activation of HOIL-1 is triggered by linear tetra-Ub binding which enables HOIL-1 to mono-ubiquitylate linear Ub chains and polysaccharides. Interestingly, the structure reveals a unique bi-nuclear Zn-cluster which substitutes the second zinc finger of the canonical RING2 fold. We identify the C-terminal histidine of this bi-nuclear Zn-cluster as the catalytic base required for the ubiquitylation activity of HOIL-1. Our study suggests that the unique zinc-coordinating architecture of RING2 provides a binding platform for ubiquitylation targets.
]]></description>
<dc:creator>Wu, Q.</dc:creator>
<dc:creator>Koliopoulos, M. G.</dc:creator>
<dc:creator>Rittinger, K.</dc:creator>
<dc:creator>Stieglitz, B.</dc:creator>
<dc:date>2022-11-13</dc:date>
<dc:identifier>doi:10.1101/2022.11.13.516300</dc:identifier>
<dc:title><![CDATA[Structural basis for ubiquitylation by HOIL-1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.11.16.516826v1?rss=1">
<title>
<![CDATA[
Cell response to extracellular matrix energy dissipation outweighs rigidity sensing 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.16.516826v1?rss=1"
</link>
<description><![CDATA[
The mechanical properties of the extracellular matrix (ECM) determine cell differentiation, proliferation and migration through mechanoresponsive proteins including YAP. However, how different mechanical signals cooperate, synergize or compete to steer cell behavior remains poorly understood. Here, we have examined competition between the two major ECM mechanical cues, i.e. rigidity, which activates cell mechanosensing, and viscous energy dissipation, which reduces stiffness blunting cell mechanotransduction. To trigger competition, we have engineered protein hydrogels allowing concomitant modulation of stiffness and viscosity by mechanisms characteristic of native ECM. Culturing cells on these hydrogels, we have found that substrate energy dissipation attenuates YAP mechanosensing prevailing over stiffness cues. Hampered YAP activation on more dissipative substrates correlates with faster actin flow and smaller focal adhesions. Mechanistically, inhibition of actomyosin contractility reverses the outcome of the competition between rigidity and energy dissipation. Our results highlight the dominating contribution of substrate viscosity to the biology of the cell.
]]></description>
<dc:creator>Huerta-Lopez, C.</dc:creator>
<dc:creator>Clemente-Manteca, A.</dc:creator>
<dc:creator>Velazquez-Carreras, D.</dc:creator>
<dc:creator>Espinosa, F. M.</dc:creator>
<dc:creator>Sanchez, J. G.</dc:creator>
<dc:creator>Saez, P.</dc:creator>
<dc:creator>Martinez-del-Pozo, A.</dc:creator>
<dc:creator>Garcia-Garcia, M.</dc:creator>
<dc:creator>Martin-Colomo, S.</dc:creator>
<dc:creator>Rodriguez-Blanco, A.</dc:creator>
<dc:creator>Esteban-Gonzalez, R.</dc:creator>
<dc:creator>Martin-Zamora, F. M.</dc:creator>
<dc:creator>Gutierrez-Rus, L.</dc:creator>
<dc:creator>Garcia, R.</dc:creator>
<dc:creator>Roca-Cusachs, P.</dc:creator>
<dc:creator>Elosegui-Artola, A.</dc:creator>
<dc:creator>del Pozo, M. A.</dc:creator>
<dc:creator>Herrero-Galan, E.</dc:creator>
<dc:creator>Plaza, G. R.</dc:creator>
<dc:creator>Alegre-Cebollada, J.</dc:creator>
<dc:date>2022-11-17</dc:date>
<dc:identifier>doi:10.1101/2022.11.16.516826</dc:identifier>
<dc:title><![CDATA[Cell response to extracellular matrix energy dissipation outweighs rigidity sensing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.11.17.516904v1?rss=1">
<title>
<![CDATA[
Whole genome error-corrected sequencing for sensitive circulating tumor DNA cancer monitoring 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.17.516904v1?rss=1"
</link>
<description><![CDATA[
Circulating cell-free DNA (ccfDNA) sequencing for low-burden cancer monitoring is limited by sparsity of circulating tumor DNA (ctDNA), the abundance of genomic material within a plasma sample, and pre-analytical error rates due to library preparation, and sequencing errors. Sequencing costs have historically favored the development of deep targeted sequencing approaches for overcoming sparsity in ctDNA detection, but these techniques are limited by the abundance of ccfDNA in samples, which imposes a ceiling on the maximal depth of coverage in targeted panels. Whole genome sequencing (WGS) is an orthogonal approach to ctDNA detection that can overcome the low abundance of ccfDNA by supplanting sequencing depth with breadth, integrating signal across the entire tumor mutation landscape. However, the higher cost of WGS limits the practical depth of coverage and hinders broad adoption. Lower sequencing costs may thus allow for enhanced ctDNA cancer monitoring via WGS. We therefore applied emerging lower-cost WGS (Ultima Genomics, 1USD/Gb) to plasma samples at [~]120x coverage. Copy number and single nucleotide variation profiles were comparable between matched Ultima and Illumina datasets, and the deeper WGS coverage enabled ctDNA detection at the parts per million range. We further harnessed these lower sequencing costs to implement duplex error-corrected sequencing at the scale of the entire genome, demonstrating a [~]1,500x decrease in errors in the plasma of patient-derived xenograft mouse models, and error rates of [~]10-7 in patient plasma samples. We leveraged this highly de-noised plasma WGS to undertake cancer monitoring in the more challenging context of resectable melanoma without matched tumor sequencing. In this context, duplex-corrected WGS allowed us to harness known mutational signature patterns for disease monitoring without matched tumors, paving the way for de novo cancer monitoring.
]]></description>
<dc:creator>Cheng, A. P.</dc:creator>
<dc:creator>Widman, A. J.</dc:creator>
<dc:creator>Arora, A.</dc:creator>
<dc:creator>Rusinek, I.</dc:creator>
<dc:creator>Hooper, W. F.</dc:creator>
<dc:creator>Murray, R.</dc:creator>
<dc:creator>Halmos, D.</dc:creator>
<dc:creator>Langanay, T.</dc:creator>
<dc:creator>Inghirami, G.</dc:creator>
<dc:creator>Germer, S.</dc:creator>
<dc:creator>Marton, M.</dc:creator>
<dc:creator>Manaa, D.</dc:creator>
<dc:creator>Helland, A.</dc:creator>
<dc:creator>Furatero, R.</dc:creator>
<dc:creator>McClintock, J.</dc:creator>
<dc:creator>Winterkorn, L.</dc:creator>
<dc:creator>Steinsnyder, Z.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Rajagopalan, S.</dc:creator>
<dc:creator>Alimohamed, A. I.</dc:creator>
<dc:creator>Malbari, M. S.</dc:creator>
<dc:creator>Saxena, A.</dc:creator>
<dc:creator>Callahan, M. K.</dc:creator>
<dc:creator>Frederick, D. T.</dc:creator>
<dc:creator>Spain, L.</dc:creator>
<dc:creator>Jaimovich, A.</dc:creator>
<dc:creator>Lipson, D.</dc:creator>
<dc:creator>Turajlic, S.</dc:creator>
<dc:creator>Zody, M. C.</dc:creator>
<dc:creator>Altorki, N. K.</dc:creator>
<dc:creator>Wolchok, J. D.</dc:creator>
<dc:creator>Postow, M. A.</dc:creator>
<dc:creator>Robine, N.</dc:creator>
<dc:creator>Boland, G.</dc:creator>
<dc:creator>Landau, D. A.</dc:creator>
<dc:date>2022-11-17</dc:date>
<dc:identifier>doi:10.1101/2022.11.17.516904</dc:identifier>
<dc:title><![CDATA[Whole genome error-corrected sequencing for sensitive circulating tumor DNA cancer monitoring]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.11.18.517032v1?rss=1">
<title>
<![CDATA[
Structural basis for Fc receptor recognition of immunoglobulin M 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.18.517032v1?rss=1"
</link>
<description><![CDATA[
FcR is the IgM-specific Fc receptor involved in the survival and activation of B cells. Using cryo-EM, we reveal eight binding sites for the human FcR Ig domain on the IgM pentamer, one of which overlaps with the receptor binding site for the transcytosis receptor pIgR, but a different mode of binding explains Ig isotype specificity. The complex explains engagement with polymeric serum IgM and the monomeric IgM B cell receptor.
]]></description>
<dc:creator>Chen, Q.</dc:creator>
<dc:creator>Menon, R.</dc:creator>
<dc:creator>Tolar, P.</dc:creator>
<dc:creator>Rosenthal, P. B.</dc:creator>
<dc:date>2022-11-18</dc:date>
<dc:identifier>doi:10.1101/2022.11.18.517032</dc:identifier>
<dc:title><![CDATA[Structural basis for Fc receptor recognition of immunoglobulin M]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.11.18.516878v1?rss=1">
<title>
<![CDATA[
Multi-site assessment of reproducibility in high-content live cell imaging data 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.18.516878v1?rss=1"
</link>
<description><![CDATA[
High-content image-based cell phenotyping provides fundamental insights in a broad variety of life science areas. Striving for accurate conclusions and meaningful impact demands high reproducibility standards, even more importantly with the advent of data sharing initiatives. However, the sources and degree of biological and technical variability, and thus the reproducibility and usefulness of meta-analysis of results from live-cell microscopy have not been systematically investigated. Here, using high content data describing features of cell migration and morphology, we determine the sources of variability across different scales, including between laboratories, persons, experiments, technical repeats, cells and time points. Significant technical variability occurred between laboratories, providing low value to direct meta-analysis on the data from different laboratories. However, batch effect removal markedly improved the possibility to combine image-based datasets of perturbation experiments. Thus, reproducible quantitative high-content cell image data and meta-analysis depend on standardized procedures and batch correction applied to studies of perturbation effects.
]]></description>
<dc:creator>Hu, J.</dc:creator>
<dc:creator>Serra-Picamal, X.</dc:creator>
<dc:creator>Bakker, G.-J.</dc:creator>
<dc:creator>Troys, M. V.</dc:creator>
<dc:creator>Winograd-katz, S.</dc:creator>
<dc:creator>Ege, N.</dc:creator>
<dc:creator>Gong, X.</dc:creator>
<dc:creator>Didan, Y.</dc:creator>
<dc:creator>Grosheva, I.</dc:creator>
<dc:creator>Polansky, O.</dc:creator>
<dc:creator>Bakkali, K.</dc:creator>
<dc:creator>Hamme, E. V.</dc:creator>
<dc:creator>Erp, M. v.</dc:creator>
<dc:creator>Vullings, M.</dc:creator>
<dc:creator>Weiss, F.</dc:creator>
<dc:creator>Clucas, J.</dc:creator>
<dc:creator>Dowbaj, A. M.</dc:creator>
<dc:creator>Sahai, E.</dc:creator>
<dc:creator>Ampe, C.</dc:creator>
<dc:creator>Geiger, B.</dc:creator>
<dc:creator>Friedl, P.</dc:creator>
<dc:creator>Bottai, M.</dc:creator>
<dc:creator>Stromblad, S.</dc:creator>
<dc:date>2022-11-20</dc:date>
<dc:identifier>doi:10.1101/2022.11.18.516878</dc:identifier>
<dc:title><![CDATA[Multi-site assessment of reproducibility in high-content live cell imaging data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.11.18.517070v1?rss=1">
<title>
<![CDATA[
The Response of the Eukaryotic Replisome to G-quadruplex- and i-motif-forming Sequences 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.18.517070v1?rss=1"
</link>
<description><![CDATA[
Sequences that can form DNA secondary structures, such as G-quadruplexes (G4s) and intercalated-Motifs (iMs), are abundant in the human genome and play a range of physiological roles. However, they can also pose a challenge to the replication machinery and in turn threaten genome stability. Multiple lines of evidence suggest G4s interfere with replication, but the underlying mechanism remains unclear. Moreover, there is a lack of evidence of how iMs affect the replisome. Here, we reconstitute replication of physiologically derived structure-forming sequences to find that a single G4 or iM is sufficient to arrest DNA replication. Direct single molecule structure detection within solid-state nanopores reveals structures form as a consequence of replication. A combination of genetic and biophysical characterisation establishes that structure forming capacity is a key determinant of replisome arrest. Mechanistically, replication fork arrest is caused by impaired synthesis, resulting in helicase-polymerase uncoupling. Significantly, iMs also induce breakage of nascent DNA. Finally, stalled forks are only rescued by a specialised helicase, Pif1, but not Sgs1 or Chl1. Altogether, this study provides a potential mechanism for quadruplex structure formation and resolution during replication and highlights G4s and iMs as endogenous sources of replication stress, which may explain their genomic instability and mutation frequencies in cancer.
]]></description>
<dc:creator>Williams, S. L.</dc:creator>
<dc:creator>Casas-Delucchi, C. S.</dc:creator>
<dc:creator>Raguseo, F.</dc:creator>
<dc:creator>Guneri, D.</dc:creator>
<dc:creator>Minamino, M.</dc:creator>
<dc:creator>Fletcher, E. E.</dc:creator>
<dc:creator>Yeeles, J. T.</dc:creator>
<dc:creator>Waller, Z. A.</dc:creator>
<dc:creator>Di Antonio, M.</dc:creator>
<dc:creator>Coster, G.</dc:creator>
<dc:date>2022-11-18</dc:date>
<dc:identifier>doi:10.1101/2022.11.18.517070</dc:identifier>
<dc:title><![CDATA[The Response of the Eukaryotic Replisome to G-quadruplex- and i-motif-forming Sequences]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.11.22.517348v1?rss=1">
<title>
<![CDATA[
Structure and function of Plasmodium actin II in the parasite mosquito stages 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.22.517348v1?rss=1"
</link>
<description><![CDATA[
Actins are filament-forming, highly-conserved proteins in eukaryotes. They are involved in essential processes in the cytoplasm and have also nuclear functions. Malaria parasites (Plasmodium spp.) have two actin isoforms that differ from each other and from canonical actins in structure and filament-forming properties. Actin I has an essential role in motility and is fairly well characterized. The structure and function of actin II are not as well understood, but mutational analyses have revealed two essential functions in male gametogenesis and in the zygote. Here, we present expression analysis, high-resolution filament structures, and biochemical characterization of Plasmodium actin II. We show that it is expressed in male gametocytes and zygotes and associated with the nucleus in both stages in filament-like structures. Unlike actin I, actin II readily forms long filaments in vitro, and near-atomic structures in the presence or absence of jasplakinolide reveal very similar structures. Small but significant differences compared to other actins in the openness and twist, the active site, the D-loop, and the plug region contribute to filament stability. Mutational analyses suggest that long and stable filaments are necessary for male gametogenesis, while the second function in the zygote stage also requires finetuned regulation by methylation of histidine 73. Actin II polymerizes via the classical nucleation-elongation mechanism and has a critical concentration of ~0.1 M at the steady-state, like actin I and canonical actins. Similarly to actin I, dimers are a stable form of actin II at equilibrium.

Significance statementMalaria is a parasitic infection caused by Plasmodium spp., which belong to the phylum Apicomplexa. In 2020, approximately 627000 people died from nearly 241 million malaria cases registered worldwide. In contrast to other apicomplexan parasites, Plasmodium spp. encode two actin isoforms. While actin I is part of the glideosome complex, essential for locomotion, actin II is present in the mosquito stages, where it has functions in gametogenesis and in the zygote. The exact function of actin II is still unclear, and information at the molecular level is limited. We show here that actin II is associated with the nucleus in gametocytes and zygotes and performs specific functions requiring both long filaments and fine-tuning by methylation of histidine 73. We determined the structures of the filamentous form of actin II at near-atomic resolution and characterized its polymerization properties in vitro. Our study provides a molecular basis for the differences between actins of the malaria parasite and humans, as well as between the two parasite actin isoforms. Furthermore, the in vivo studies provide insights into the function of actin II in the parasite.
]]></description>
<dc:creator>Lopez, A. J.</dc:creator>
<dc:creator>Andreadaki, M.</dc:creator>
<dc:creator>Vahokoski, J.</dc:creator>
<dc:creator>Deligianni, E.</dc:creator>
<dc:creator>Calder, L. J.</dc:creator>
<dc:creator>Camerini, S.</dc:creator>
<dc:creator>Freitag, A.</dc:creator>
<dc:creator>Bergmann, U.</dc:creator>
<dc:creator>Rosenthal, P. B.</dc:creator>
<dc:creator>Siden-Kiamos, I.</dc:creator>
<dc:creator>Kursula, I.</dc:creator>
<dc:date>2022-11-24</dc:date>
<dc:identifier>doi:10.1101/2022.11.22.517348</dc:identifier>
<dc:title><![CDATA[Structure and function of Plasmodium actin II in the parasite mosquito stages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.11.24.517538v1?rss=1">
<title>
<![CDATA[
Epilepsy-linked kinase CDKL5 phosphorylates voltage-gated calcium channel Cav2.3, altering inactivation kinetics and neuronal excitability 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.24.517538v1?rss=1"
</link>
<description><![CDATA[
Developmental and epileptic encephalopathies (DEEs) are a group of rare childhood disorders characterized by severe epilepsy and cognitive deficits. Numerous DEE genes have been discovered thanks to advances in genomic diagnosis, yet putative molecular links between these disorders are unknown. CDKL5 deficiency disorder (CDD, DEE2), one of the most common genetic epilepsies, is caused by loss-of-function mutations in the brain-enriched kinase CDKL5. To elucidate CDKL5 function, we looked for CDKL5 substrates using a SILAC-based phosphoproteomic screen. We identified the voltage-gated Ca2+ channel Cav2.3 (encoded by CACNA1E) as a novel physiological target of CDKL5 in mice and humans. Recombinant channel electrophysiology and interdisciplinary characterization of Cav2.3 phosphomutant mice revealed that loss of Cav2.3 phosphorylation leads to channel gain-of-function via slower inactivation and enhanced cholinergic stimulation, resulting in increased neuronal excitability. Our results thus show that CDD is partly a channelopathy. The properties of unphosphorylated Cav2.3 closely resemble those described for CACNA1E gain-of-function mutations causing DEE69, a disorder sharing clinical features with CDD. We show that these two single-gene diseases are mechanistically related and could be ameliorated with Cav2.3 inhibitors.
]]></description>
<dc:creator>Sampedro-Castaneda, M.</dc:creator>
<dc:creator>Baltussen, L. L.</dc:creator>
<dc:creator>Lopes, A. T.</dc:creator>
<dc:creator>Qiu, Y.</dc:creator>
<dc:creator>Sirvio, L.</dc:creator>
<dc:creator>Mihaylov, S. R.</dc:creator>
<dc:creator>Claxton, S.</dc:creator>
<dc:creator>Richardson, J. C.</dc:creator>
<dc:creator>Lignani, G.</dc:creator>
<dc:creator>Ultanir, S. K.</dc:creator>
<dc:date>2022-11-24</dc:date>
<dc:identifier>doi:10.1101/2022.11.24.517538</dc:identifier>
<dc:title><![CDATA[Epilepsy-linked kinase CDKL5 phosphorylates voltage-gated calcium channel Cav2.3, altering inactivation kinetics and neuronal excitability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.07.15.500256v1?rss=1">
<title>
<![CDATA[
Rarity: Discovering rare cell populations from single-cell imaging data 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.07.15.500256v1?rss=1"
</link>
<description><![CDATA[
BackgroundCell type identification plays an important role in the analysis and interpretation of single-cell data and can be carried out via supervised or unsupervised clustering approaches. Supervised methods are best suited where we can list all cell types and their respective marker genes a priori. While unsupervised clustering algorithms look for groups of cells with similar expression properties. This property permits the identification of both known and unknown cell populations, making unsupervised methods suitable for discovery.. Success is dependent on the relative strength of the expression signature of each group as well as the number of cells. Rare cell types therefore present a particular challenge that are magnified when they are defined by differentially expressing a small number of genes. Typical unsupervised approaches fail to identify such rare subpopulations, and these cells tend to be absorbed into more prevalent cell types.

ResultsIn order to balance these competing demands, we have developed a novel statistical framework for unsupervised clustering, named Rarity, that enables the discovery process for rare cell types to be more robust, consistent and interpretable. We achieve this by devising a novel clustering method based on a Bayesian latent variable model in which we assign cells to inferred latent binary on/off expression profiles. This lets us achieve increased sensitivity to rare cell populations while also allowing us to control and interpret potential false positive discoveries.

ConclusionsWe systematically study the challenges associated with rare cell type identification and demonstrate the utility of Rarity on various IMC data sets.
]]></description>
<dc:creator>Marten, K.</dc:creator>
<dc:creator>Bortolomeazzi, M.</dc:creator>
<dc:creator>Montorsi, L.</dc:creator>
<dc:creator>Spencer, J.</dc:creator>
<dc:creator>Ciccarelli, F. D.</dc:creator>
<dc:creator>Yau, C.</dc:creator>
<dc:date>2022-07-18</dc:date>
<dc:identifier>doi:10.1101/2022.07.15.500256</dc:identifier>
<dc:title><![CDATA[Rarity: Discovering rare cell populations from single-cell imaging data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.11.30.518495v1?rss=1">
<title>
<![CDATA[
Tsetse flies choose birthing sites guided by environmental but not pheromonal cues 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.30.518495v1?rss=1"
</link>
<description><![CDATA[
Tsetse flies significantly impact public health and economic development in sub-Saharan African countries by transmitting the fatal disease African trypanosomiasis. Unusually, instead of laying eggs, tsetse birth a single larva that immediately burrows into the soil to pupate. Where the female chooses to larviposit is therefore crucial for offspring survival. Previous studies showed that a putative larval pheromone, n-pentadecane, attracts gravid female Glossina morsitans morsitans to appropriate larviposition sites. However, this attraction could not be reproduced in field experiments. Here, we resolve this disparity by designing naturalistic laboratory experiments that closely mimic the characteristics found in the wild. We show that gravid tsetse were neither attracted to the putative pheromone nor, interestingly, to pupae placed in the soil. In contrast, females appear to choose larviposition sites based on environmental cues. We conclude that it is the substrate, rather than larval pheromones, which drives larviposition site selection under naturalistic conditions.
]]></description>
<dc:creator>Adden, A. K.</dc:creator>
<dc:creator>Haines, L. R.</dc:creator>
<dc:creator>Acosta-Serrano, A.</dc:creator>
<dc:creator>Prieto-Godino, L. L.</dc:creator>
<dc:date>2022-12-01</dc:date>
<dc:identifier>doi:10.1101/2022.11.30.518495</dc:identifier>
<dc:title><![CDATA[Tsetse flies choose birthing sites guided by environmental but not pheromonal cues]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.12.01.518339v1?rss=1">
<title>
<![CDATA[
A role for MCH neuron firing in hippocampal plasticity and learning 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.01.518339v1?rss=1"
</link>
<description><![CDATA[
It has been revealed that melanin-concentrating hormone (MCH) neurons in the hypothalamus can influence learning (Liu et al., 2022) and memory formation (Kosse & Burdakov, 2019), but the cellular mechanisms by which they perform this function are not understood. Here, we examine the role of MCH neural input to the hippocampus, and show in vitro that optogenetically increasing MCH axon activity facilitates hippocampal plasticity by lowering the threshold for synaptic potentiation. In vivo, we find that MCH neurons are naturally active in response to reinforcing cues during a spatial learning task, and that this activity is correlated with the speed of learning. Together, our results align with increasing evidence that MCH neurons play an  on-line regulatory role in learning, and reveal that this could be achieved through modulation of synaptic plasticity in the hippocampus.
]]></description>
<dc:creator>Harris, J. J.</dc:creator>
<dc:creator>Concetti, C.</dc:creator>
<dc:creator>Peleg-Raibstein, D.</dc:creator>
<dc:creator>Burdakov, D.</dc:creator>
<dc:date>2022-12-01</dc:date>
<dc:identifier>doi:10.1101/2022.12.01.518339</dc:identifier>
<dc:title><![CDATA[A role for MCH neuron firing in hippocampal plasticity and learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.12.03.518823v1?rss=1">
<title>
<![CDATA[
Investigating the genetic diversity of H5 avian influenza in the UK 2020-2022 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.03.518823v1?rss=1"
</link>
<description><![CDATA[
Since 2020, the UK and Europe, have experienced annual epizootics of high pathogenicity avian influenza virus (HPAIV). The first during autumn/winter 2020/21 involved the detected with six H5Nx subtypes although H5N8 HPAIV dominated in the UK. Whilst genetic assessment of the H5N8 HPAIVs within the UK demonstrated relative homogeneity, there was a background of other genotypes circulating at a lower degree with different neuraminidase and internal genes. Following a small number of summer detections of H5N1 in wild birds over the summer of 2021, autumn/winter 2021/22 saw another European H5 HPAIV epizootic, that has dwarfed the prior epizootic. This second epizootic was dominated almost exclusively by H5N1 HPAIV, although six distinct genotypes were defined. We have used genetic analysis to evaluate the emergence of different genotypes and proposed reassortment events that have been observed. The existing data suggests that the H5N1 circulating in Europe during late 2020, continued to circulate in wild birds throughout 2021, with minimal adaptation, but has then gone on to reassort with AIVs in the wild bird population. We have undertaken an in-depth genetic assessment of H5 HPAIVs detected in the UK, over the last two winter seasons and demonstrate the utility of in-depth genetic analyses in defining the diversity of H5 HPAIVs circulating in avian species, the potential for zoonotic risk and whether incidents of lateral spread can be defined over independent incursion of infection from wild birds. Key supporting data for mitigation activities.

ImportanceHigh pathogenicity avian influenza virus (HPAIV) outbreaks devastate avian species across all sectors having both economic and ecological impacts through mortalities in poultry and wild birds, respectively. These viruses can also represent a significant zoonotic risk. Since 2020, the UK has experienced two successive outbreaks of H5 HPAIV. Whilst H5N8 HPAIV was predominant during the 2020/21 outbreak, other H5 subtypes were also detected. The following year there was a shift in subtype dominance to H5N1 HPAIV, but multiple H5N1 genotypes were detected. Through thorough utilisation of whole-genome sequencing, it was possible to track and characterise the genetic evolution of these H5 HPAIVs in UK poultry and wild birds. This has enabled us to assess the risk posed by these viruses at the poultry:wild bird and the avian:human interface and to investigate potential lateral spread between infected premises, a key factor in understanding threat to the commercial sector.
]]></description>
<dc:creator>Byrne, A. M.</dc:creator>
<dc:creator>James, J.</dc:creator>
<dc:creator>Mollett, B. C.</dc:creator>
<dc:creator>Meyer, S. M.</dc:creator>
<dc:creator>Lewis, T.</dc:creator>
<dc:creator>Czepiel, M.</dc:creator>
<dc:creator>Seekings, A. H.</dc:creator>
<dc:creator>Mahmood, S.</dc:creator>
<dc:creator>Thomas, S. S.</dc:creator>
<dc:creator>Ross, C. S.</dc:creator>
<dc:creator>Byrne, D. J.</dc:creator>
<dc:creator>McMenamy, M. J.</dc:creator>
<dc:creator>Bailie, V.</dc:creator>
<dc:creator>Lemon, K.</dc:creator>
<dc:creator>Hansen, R. D.</dc:creator>
<dc:creator>Falchieri, M.</dc:creator>
<dc:creator>Lewis, N. S.</dc:creator>
<dc:creator>Reid, S. M.</dc:creator>
<dc:creator>Brown, I. H.</dc:creator>
<dc:creator>Banyard, A. C.</dc:creator>
<dc:date>2022-12-03</dc:date>
<dc:identifier>doi:10.1101/2022.12.03.518823</dc:identifier>
<dc:title><![CDATA[Investigating the genetic diversity of H5 avian influenza in the UK 2020-2022]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.12.08.519568v1?rss=1">
<title>
<![CDATA[
A heterotrimeric complex of Toxoplasma proteins promotes parasite survival in interferon gamma stimulated human cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.08.519568v1?rss=1"
</link>
<description><![CDATA[
Toxoplasma gondii secretes protein effectors to subvert the human immune system sufficiently to establish a chronic infection. Relative to murine infections, little is known about which parasite effectors disarm human immune responses. Here we used targeted CRISPR screening to identify secreted protein effectors required for parasite survival in IFN{gamma}-activated human cells. Independent screens were carried out using two Toxoplasma strains which differ in virulence in mice, leading to the identification of effectors required for survival in IFN{gamma}-activated human cells. We identify the secreted protein GRA57 and two other proteins, GRA70 and GRA71, that together form a complex which enhances the ability of parasites to persist in IFN{gamma}-activated human foreskin fibroblasts (HFFs). Components of the protein machinery required for export of Toxoplasma proteins into the host cell were also found to be important for parasite resistance to IFN{gamma} in human cells, but these export components function independently of the identified protein complex. Host-mediated ubiquitination of the parasite vacuole has previously been associated with increased parasite clearance from human cells, but we find that vacuoles from GRA57, GRA70 and GRA71 knockout strains are surprisingly less ubiquitinated by the host cell. We hypothesise that deletion of this trimeric complex renders parasites hypersensitive to remaining ubiquitination, resulting in increased parasite clearance.
]]></description>
<dc:creator>Lockyer, E.</dc:creator>
<dc:creator>Torelli, F.</dc:creator>
<dc:creator>Butterworth, S.</dc:creator>
<dc:creator>Song, O.-R.</dc:creator>
<dc:creator>Howell, S.</dc:creator>
<dc:creator>Weston, A.</dc:creator>
<dc:creator>East, P.</dc:creator>
<dc:creator>Treeck, M.</dc:creator>
<dc:date>2022-12-08</dc:date>
<dc:identifier>doi:10.1101/2022.12.08.519568</dc:identifier>
<dc:title><![CDATA[A heterotrimeric complex of Toxoplasma proteins promotes parasite survival in interferon gamma stimulated human cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.12.09.519777v1?rss=1">
<title>
<![CDATA[
p38γ and p38δ modulate innate immune response by regulating MEF2D activation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.09.519777v1?rss=1"
</link>
<description><![CDATA[
Evidence implicating p38{gamma} and p38{delta} (p38{gamma}/p38{delta}) in inflammation are mainly based on experiments using p38{gamma}/p38{delta} deficient (p38{gamma}/{delta}-/-) mice, which show low levels of TPL2, the kinase upstream of MKK1-ERK1/2 in myeloid cells. This could obscure p38{gamma}/p38{delta} roles, since TPL2 is essential for regulating inflammation. Here we generated a p38{gamma}D171A/D171A/p38{delta}-/- (p38{gamma}/{delta}KIKO) mouse, expressing kinase-inactive p38{gamma} and lacking p38{delta}. This mouse exhibited normal TPL2 levels, making it an excellent tool to elucidate specific p38{gamma}/p38{delta} functions. p38{gamma}/{delta}KIKO mice showed a reduced inflammatory response and less susceptibility to LPS-induced septic shock and Candida albicans infection than wild-type mice. Gene expression analyses in LPS-activated WT and p38{gamma}/{delta}KIKO macrophages revealed that p38{gamma}/p38{delta} regulated numerous genes implicated in innate immune response. Additionally, phospho-proteomic analyses and in vitro kinase assays showed that the transcription factor myocyte enhancer factor-2D (MEF2D) was phosphorylated at Ser444 via p38{gamma}/p38{delta}. Mutation of MEF2D Ser444 to the non-phosphorylatable residue Ala increased its transcriptional activity and the expression of iNOS and IL-1{beta} mRNA. These results suggest that p38{gamma}/p38{delta} govern innate immune responses by regulating MEF2D phosphorylation and transcriptional activity.
]]></description>
<dc:creator>Cuenda, A.</dc:creator>
<dc:creator>Escos, A.</dc:creator>
<dc:creator>Diaz-Mora, E.</dc:creator>
<dc:creator>Fajardo, P.</dc:creator>
<dc:creator>Gonzalez-Romero, D.</dc:creator>
<dc:creator>Risco, A.</dc:creator>
<dc:creator>Martin-Gomez, J.</dc:creator>
<dc:creator>Bonneil, E.</dc:creator>
<dc:creator>Sonenberg, N.</dc:creator>
<dc:creator>Jafarnejad, S. M.</dc:creator>
<dc:creator>Pattison, M.</dc:creator>
<dc:creator>Sanz-Ezquerro, J. J.</dc:creator>
<dc:creator>Ley, S. C.</dc:creator>
<dc:date>2022-12-09</dc:date>
<dc:identifier>doi:10.1101/2022.12.09.519777</dc:identifier>
<dc:title><![CDATA[p38γ and p38δ modulate innate immune response by regulating MEF2D activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.12.09.519741v1?rss=1">
<title>
<![CDATA[
Structure of Toxoplasma gondii glideosome-associated connector suggests a role as an elastic element in actomyosin force generation for gliding motility 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.09.519741v1?rss=1"
</link>
<description><![CDATA[
Toxoplasma gondii glideosome-associated connector (GAC) is a giant armadillo-repeat protein, essential for parasite motility and conserved across Apicomplexa. It connects actin filaments to the plasma membrane via interactions with phosphatidic acid and membrane-spanning adhesins. It is unclear how GAC contributes to gliding motility and invasion and why such a large connector is needed. We determined the crystal structure of full-length T. gondii GAC at 2.3 [A] resolution and explored its conformational space in solution using small-angle X-ray scattering and cryogenic electron microscopy. The crystal structure reveals a compact conformation but, in solution, GAC adopts both compact and extended forms. The PH domain stabilizes the compact form and may act as a switch triggered by membrane sensing. Based on its spring-like architecture, we suggest a role for GAC as an elastic element in actomyosin force generation during gliding motility and invasion.
]]></description>
<dc:creator>Hung, Y.-F.</dc:creator>
<dc:creator>Chen, Q.</dc:creator>
<dc:creator>Pires, I.</dc:creator>
<dc:creator>Rosenthal, P. B.</dc:creator>
<dc:creator>Kursula, I.</dc:creator>
<dc:date>2022-12-10</dc:date>
<dc:identifier>doi:10.1101/2022.12.09.519741</dc:identifier>
<dc:title><![CDATA[Structure of Toxoplasma gondii glideosome-associated connector suggests a role as an elastic element in actomyosin force generation for gliding motility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.12.20.521107v1?rss=1">
<title>
<![CDATA[
The paralogues MAGOH and MAGOHB are oncogenic factors in high-grade gliomas and safeguard the splicing of cell division and cell cycle genes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.20.521107v1?rss=1"
</link>
<description><![CDATA[
The exon junction complex (EJC) plays key roles throughout the lifespan of RNA and is particularly relevant in the nervous system. We investigated the roles of two EJC members, the paralogs MAGOH and MAGOHB, with respect to brain tumor development. High MAGOH/MAGOHB expression was observed in 14 tumor types; glioblastoma (GBM) showed the greatest difference compared to normal tissue. Increased MAGOH/MAGOHB expression was associated with poor prognosis in glioma patients, while knockdown of MAGOH/MAGOHB affected different cancer phenotypes. Reduced MAGOH/MAGOHB expression in GBM cells caused alterations in the splicing profile, including re-splicing and skipping of multiple exons. The binding profiles of EJC proteins indicated that exons affected by MAGOH/MAGOHB knockdown accumulated fewer complexes on average, providing a possible explanation for their sensitivity to MAGOH/MAGOHB knockdown. Transcripts (genes) showing alterations in the splicing profile are mainly implicated in cell division, cell cycle, splicing, and translation. We propose that high MAGOH/MAGOHB levels are required to safeguard the splicing of genes in high demand in scenarios requiring increased cell proliferation (brain development and GBM growth), ensuring efficient cell division, cell cycle regulation, and gene expression (splicing and translation). Since differentiated neuronal cells do not require increased MAGOH/MAGOHB expression, targeting these paralogs is a potential option for treating GBM.
]]></description>
<dc:creator>Barreiro, R. A. S.</dc:creator>
<dc:creator>Guardia, G. D. A.</dc:creator>
<dc:creator>Meliso, F. M.</dc:creator>
<dc:creator>Lei, X.</dc:creator>
<dc:creator>Savio, A.</dc:creator>
<dc:creator>Fellermeyer, M.</dc:creator>
<dc:creator>Conceicao, H. B.</dc:creator>
<dc:creator>Mercuri, R. L. V.</dc:creator>
<dc:creator>Landry, T.</dc:creator>
<dc:creator>Qiao, M.</dc:creator>
<dc:creator>Blazquez, L.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Penalva, L. O.</dc:creator>
<dc:creator>Galante, P. A. F.</dc:creator>
<dc:date>2022-12-20</dc:date>
<dc:identifier>doi:10.1101/2022.12.20.521107</dc:identifier>
<dc:title><![CDATA[The paralogues MAGOH and MAGOHB are oncogenic factors in high-grade gliomas and safeguard the splicing of cell division and cell cycle genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.12.21.521417v1?rss=1">
<title>
<![CDATA[
Back to the Future: Unleashing your cytometers spectral potential 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.21.521417v1?rss=1"
</link>
<description><![CDATA[
With the recent growth in spectral flow cytometry many laboratories are investing in new spectral flow cytometers in order to maximise the information gathered about every cell. This study hypothesised that traditional cytometers already within many laboratories may be used as spectral cytometers and have shown using a range of different cytometers that data acquired may be unmixed after acquisition.
]]></description>
<dc:creator>Walker, R. V.</dc:creator>
<dc:creator>Hall, C.</dc:creator>
<dc:creator>Ibrahim, H.</dc:creator>
<dc:creator>Thompson, S.</dc:creator>
<dc:creator>Hobson, P.</dc:creator>
<dc:creator>Crofts, J.-A.</dc:creator>
<dc:creator>Nobes, P.</dc:creator>
<dc:creator>Lim, S.</dc:creator>
<dc:creator>Burpee, T.</dc:creator>
<dc:date>2022-12-22</dc:date>
<dc:identifier>doi:10.1101/2022.12.21.521417</dc:identifier>
<dc:title><![CDATA[Back to the Future: Unleashing your cytometers spectral potential]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.12.22.521600v1?rss=1">
<title>
<![CDATA[
Identification of novel coloboma candidate genes through conserved gene expression analyses across four vertebrate species 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.22.521600v1?rss=1"
</link>
<description><![CDATA[
Ocular coloboma (OC) is a failure of complete optic fissure closure during embryonic development and presents as a tissue defect along the proximal distal axis of the ventral eye. It is classed as part of the clinical spectrum of structural eye malformations with microphthalmia and anophthalmia, collectively abbreviated to MAC. Despite deliberate attempts to identify causative variants in MAC, many patients remain without a genetic diagnosis. To reveal potential candidate genes, we utilised transcriptomes experimentally generated from embryonic eye tissues derived from human, mouse, zebrafish, and chicken at stages coincident with optic fissure closure. Our in-silico analyses found 10 genes with optic fissure specific enriched expression: ALDH1A3, BMPR1B, EMX2, EPHB3, NID1, NTN1, PAX2, SMOC1, TENM3, and VAX1. In situ hybridization revealed that all 10 genes were broadly expressed ventrally in the developing eye, but that only PAX2 and NTN1 were expressed in cells at the edges of the optic fissure margin. Of these conserved optic fissure genes, EMX2, NID1, and EPHB3 have not previously been associated with human MAC cases. Targeted genetic manipulation in zebrafish embryos using CRISPR/Cas9 caused the developmental MAC phenotype for emx2 and ephb3. We scrutined available whole genome sequencing datasets from MAC patients and identified a range of variants with plausible causality. In combination our data suggest that expression of genes involved in ventral eye development are conserved across a range of vertebrate species, and that EMX2, NID1, and EPHB3 are candidate loci that should be adopted into clinical diagnostic screens for patients with structural eye malformations.
]]></description>
<dc:creator>Trejo-Reveles, V.</dc:creator>
<dc:creator>Owen, N.</dc:creator>
<dc:creator>Chan, B. H. C.</dc:creator>
<dc:creator>Toms, M.</dc:creator>
<dc:creator>Schoenebeck, J. J.</dc:creator>
<dc:creator>Moosajee, M.</dc:creator>
<dc:creator>Rainger, J.</dc:creator>
<dc:date>2022-12-22</dc:date>
<dc:identifier>doi:10.1101/2022.12.22.521600</dc:identifier>
<dc:title><![CDATA[Identification of novel coloboma candidate genes through conserved gene expression analyses across four vertebrate species]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.12.22.521633v1?rss=1">
<title>
<![CDATA[
Cleavage furrow-directed cortical flows bias mechanochemical pathways for PAR polarization in the C. elegans germ lineage 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.22.521633v1?rss=1"
</link>
<description><![CDATA[
During development, the conserved PAR polarity network is continuously redeployed, requiring that it adapts to changing cellular contexts and environmental cues. How it does so and the degree to which these adaptations reflect changes in its fundamental design principles remain unclear. Here, we investigate the process of PAR polarization within the highly tractable C. elegans germline P lineage, which undergoes a series of iterative asymmetric stem cell-like divisions. Compared to the zygote, we observe significant differences in the pattern of polarity emergence, including an inversion of the initial unpolarized state, changes in symmetry breaking cues, and the timings with which anterior and posterior PARs segregate. Beneath these differences, however, polarity establishment remains reliant on the same core pathways identified in the zygote, including conserved roles for cortical actin flows and PAR-dependent self-organization. Intriguingly, we find that cleavage furrow-directed cortical actin flows play a similar symmetry-breaking role for the germline cell P1 as centrosome-induced cortical flows in the zygote. Through their ability to induce asymmetric accumulation of PAR-3 clusters, these furrow-directed flows directly couple the geometry of polarization to cell division, which could be a general strategy for cells to ensure proper organization within dynamically growing systems, such as embryos. In summary, our data suggest that coupling of novel symmetry-breaking cues with highly adaptable core mechanochemical circuits enable robust PAR polarity in response to changing developmental contexts.
]]></description>
<dc:creator>Ng, K.</dc:creator>
<dc:creator>Hirani, N.</dc:creator>
<dc:creator>Bland, T.</dc:creator>
<dc:creator>Pinto, J. B.</dc:creator>
<dc:creator>Goehring, N. W.</dc:creator>
<dc:date>2022-12-23</dc:date>
<dc:identifier>doi:10.1101/2022.12.22.521633</dc:identifier>
<dc:title><![CDATA[Cleavage furrow-directed cortical flows bias mechanochemical pathways for PAR polarization in the C. elegans germ lineage]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.12.23.521684v1?rss=1">
<title>
<![CDATA[
Nucleotide binding halts diffusion of the eukaryotic replicative helicase during activation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.23.521684v1?rss=1"
</link>
<description><![CDATA[
The eukaryotic replicative helicase CMG centrally orchestrates the replisome and leads the way at the front of replication forks1. Understanding the motion of CMG on the DNA is therefore key to our understanding of DNA replication. In vivo, CMG is assembled and activated through a cell-cycle-regulated mechanism involving 36 polypeptides that has been reconstituted from purified proteins in ensemble biochemical studies2,3. Conversely, single-molecule studies of CMG motion have thus far4-6 relied on pre-formed CMG assembled through an unknown mechanism upon overexpression of individual constituents7,8. Here, we report the first activation at the single-molecule level of CMG fully reconstituted from purified yeast proteins and the quantification of its motion. We observe that CMG can move on DNA in two ways: by unidirectional translocation and by diffusion. We demonstrate that CMG preferentially exhibits unidirectional translocation in the presence of ATP, whereas it preferentially exhibits diffusive motion in the absence of ATP. We also demonstrate that nucleotide binding halts diffusive CMG. Taken together, our findings support a mechanism by which nucleotide binding allows newly assembled CMG to engage with the DNA within its central channel without melting it, halting its diffusion and facilitating the initial DNA melting required to initiate DNA replication.
]]></description>
<dc:creator>Ramirez Montero, D.</dc:creator>
<dc:creator>Sanchez, H.</dc:creator>
<dc:creator>van Veen, E.</dc:creator>
<dc:creator>van Laar, T.</dc:creator>
<dc:creator>Solano, B.</dc:creator>
<dc:creator>Diffley, J. F. X.</dc:creator>
<dc:creator>Dekker, N. H.</dc:creator>
<dc:date>2022-12-23</dc:date>
<dc:identifier>doi:10.1101/2022.12.23.521684</dc:identifier>
<dc:title><![CDATA[Nucleotide binding halts diffusion of the eukaryotic replicative helicase during activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.12.25.521885v1?rss=1">
<title>
<![CDATA[
Discovery and characterization of cyclic peptides selective for the C-terminal bromodomains of BET family proteins 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.25.521885v1?rss=1"
</link>
<description><![CDATA[
SUMMARYDNA encoded cyclic peptide libraries offer unique opportunities to discover high-potency, high-specificity ligands directed against a target protein. We set out to explore the potential for such libraries to provide ligands that can distinguish between bromodomains from the closely related paralogues of the Bromodomain and ExtraTerminal domain (BET) family of epigenetic regulators. Analysis of peptides isolated from a screen against the C-terminal bromodomain of family member BRD2, together with new peptides discovered in previous screens against the corresponding domain from BRD3 and BRD4, reveals peptides with nanomolar and subnanomolar affinities. X-ray crystal structures of several of these bromodomain-peptide complexes reveal diverse structures and binding modes, which nevertheless display several conserved binding features. A subset of the peptides demonstrates significant paralogue-level specificity, though structural analysis does not reveal clear physicochemical explanations for this specificity. Our data demonstrate the power of cyclic peptides to discriminate between highly similar proteins with high potency and hint that differences in conformational dynamics between BET-family bromodomains might modulate binding affinities amongst family members for particular ligands.
]]></description>
<dc:creator>Franck, C.</dc:creator>
<dc:creator>Patel, K.</dc:creator>
<dc:creator>Walport, L. J.</dc:creator>
<dc:creator>Christie, M. T.</dc:creator>
<dc:creator>Norman, A.</dc:creator>
<dc:creator>Passioura, T.</dc:creator>
<dc:creator>Payne, R. J.</dc:creator>
<dc:creator>Suga, H.</dc:creator>
<dc:creator>Mackay, J. P.</dc:creator>
<dc:date>2022-12-26</dc:date>
<dc:identifier>doi:10.1101/2022.12.25.521885</dc:identifier>
<dc:title><![CDATA[Discovery and characterization of cyclic peptides selective for the C-terminal bromodomains of BET family proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.12.29.521787v1?rss=1">
<title>
<![CDATA[
A practical extraction and spatial statistical pipeline for large 3D bioimages 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.29.521787v1?rss=1"
</link>
<description><![CDATA[
ABSTRACT/SUMMARYDespite much being known about the molecular regulation of hematopoiesis, little is understood about how hematopoietic cells are organized within bone marrow (BM) tissue. Recent advances in microscopy have led to the creation of increasingly detailed images of murine hematopoietic tissue. Accurate, efficient, and informative methodologies to extract and analyze the large amount of data generated are, however, still lacking. Indeed, cells are very densely packed in the bone marrow and therefore difficult to efficiently and accurately segment. In addition, currently employed statical analyses of cellular localization are generally unsuitable for the investigation of interactions between more than two cell types. To overcome these limitations, we developed PACESS, a readily applicable method based on neural network classification of hundreds of thousands of cells in thick 3D bone marrow samples, and a combination of statistical techniques to assess the spatial interactions between multiple cell types. To validate this approach, we used it to investigate the spatial organization of T cells, megakaryocytes and leukemic cells. We demonstrate that the presence of large clusters of leukemic cells affects the distribution of both T cells and megakaryocytes, albeit differently, resulting in the generation of previously unrecognized, unique microenvironments adjacent to each other within the same bone marrow cavity. We believe that this approach can contribute to unravel the BM cellular organization.

MOTIVATIONThe organization of diverse hematopoietic cells within bone marrow tissue remains unclear. Recently developed tissue clearing methods enable the generation of large, 3D, single cell resolution microscopy images datasets, but the bottleneck in their analysis lies in both the identification and classification of cells, and in statistical analyses to probe their spatial relationships. We present a workflow (PACESS) that takes advantage of a convoluted neural network to identify and classify cells in 2D coupled with an automated method that extrapolates to 3D, followed by a combination of spatial statistics to classify tissue regions based on each cell types density, and logistic regression to test whether the relative abundance of cell types may be explained by reciprocal dependencies. Finally, we provide a combined measurement of the abundance of all cell types in a 3D map, highlighting regional variations found within the tissue.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=195 SRC="FIGDIR/small/521787v3_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Adams, G.</dc:creator>
<dc:creator>Tissot, F.</dc:creator>
<dc:creator>Liu, C.</dc:creator>
<dc:creator>Brunsdon, C.</dc:creator>
<dc:creator>Duffy, K.</dc:creator>
<dc:creator>Lo Celso, C.</dc:creator>
<dc:date>2022-12-29</dc:date>
<dc:identifier>doi:10.1101/2022.12.29.521787</dc:identifier>
<dc:title><![CDATA[A practical extraction and spatial statistical pipeline for large 3D bioimages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.12.29.522219v1?rss=1">
<title>
<![CDATA[
Optimization of energy production and central carbon metabolism in a non-respiring eukaryote 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.29.522219v1?rss=1"
</link>
<description><![CDATA[
Most eukaryotes respire oxygen, using it to generate biomass and energy. Yet, a few organisms lost the capacity to respire. Understanding how they manage biomass and energy production may illuminate the critical points at which respiration feeds into central carbon metabolism and explain possible routes to its optimization. Here we use two related fission yeasts, Schizosaccharomyces pombe and Schizosaccharomyces japonicus, as a comparative model system. We show that although S. japonicus does not respire oxygen, unlike S. pombe, it is capable of efficient NADH oxidation, amino acid synthesis and ATP generation. We probe possible optimization strategies using stable isotope tracing metabolomics, mass isotopologue distribution analysis, genetics, and physiological experiments. S. japonicus appears to have optimized cytosolic NADH oxidation via glycerol-3-phosphate synthesis. It runs a fully bifurcated TCA  cycle, supporting higher amino acid production. Finally, it uses the pentose phosphate pathway both to support faster biomass generation and as a shunt to optimize glycolytic flux, thus producing more ATP than the respiro-fermenting S. pombe. By comparing two related organisms with vastly different metabolic strategies, our work highlights the versatility and plasticity of central carbon metabolism in eukaryotes, illuminating critical adaptations supporting the preferential use of glycolysis over oxidative phosphorylation.
]]></description>
<dc:creator>Alam, S.</dc:creator>
<dc:creator>Gu, Y.</dc:creator>
<dc:creator>Reichert, P.</dc:creator>
<dc:creator>Bahler, J.</dc:creator>
<dc:creator>Oliferenko, S.</dc:creator>
<dc:date>2022-12-29</dc:date>
<dc:identifier>doi:10.1101/2022.12.29.522219</dc:identifier>
<dc:title><![CDATA[Optimization of energy production and central carbon metabolism in a non-respiring eukaryote]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.03.522491v1?rss=1">
<title>
<![CDATA[
Control of atypical PKC{iota} membrane dissociation by tyrosine phosphorylation within a PB1-C1 interdomain interface 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.03.522491v1?rss=1"
</link>
<description><![CDATA[
Atypical PKCs are cell polarity kinases that operate at the plasma membrane where they function within multiple molecular complexes to contribute to the establishment and maintenance of polarity. In contrast to the classical and novel PKCs, atypical PKCs do not respond to diacylglycerol cues to bind the membrane compartment. Until recently it was not clear how aPKCs are recruited; whether aPKCs can directly interact with membranes or whether they are dependent on other protein interactors to do so. Two recent studies identified the pseudo-substrate region and the C1 domain as direct membrane interaction modules, however their relative importance and coupling are unknown. We combined molecular modelling and functional assays to show that the regulatory module of aPKC{iota}, comprising the PB1 pseudo-substrate and C1 domains, forms a cooperative and spatially continuous invariant membrane interaction platform. Furthermore, we show the coordinated orientation of membrane-binding elements within the regulatory module requires a key PB1-C1 interfacial {beta}-strand (BSL). We show this element contains a highly conserved Tyr residue that can be phosphorylated and that negatively regulates the integrity of the regulatory module, leading to membrane release. We thus expose a novel regulatory mechanism of aPKC{iota} membrane binding and release during cell polarization.
]]></description>
<dc:creator>Cobbaut, M.</dc:creator>
<dc:creator>McDonald, N. Q.</dc:creator>
<dc:creator>Parker, P. J.</dc:creator>
<dc:date>2023-01-03</dc:date>
<dc:identifier>doi:10.1101/2023.01.03.522491</dc:identifier>
<dc:title><![CDATA[Control of atypical PKC{iota} membrane dissociation by tyrosine phosphorylation within a PB1-C1 interdomain interface]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.06.521078v1?rss=1">
<title>
<![CDATA[
Genomic evolution of non-small cell lung cancer patient-derived xenograft models 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.06.521078v1?rss=1"
</link>
<description><![CDATA[
Patient-derived xenograft (PDX) models of cancer, developed through injection of patient tumour cells into immunocompromised mice, have been widely adopted in preclinical studies, as well as in precision oncology approaches. However, the extent to which PDX models represent the underlying genetic diversity of a patients tumour and the extent of on-going genomic evolution in PDX models are incompletely understood, particularly in the context of heterogeneous cancers such as non-small cell lung cancer (NSCLC). To investigate the depiction of intratumour heterogeneity by PDX models, we derived 47 new subcutaneous multi-region PDX models from 22 patients with primary NSCLC enrolled in the clinical longitudinal cohort study TRACERx. By analysing whole exome sequencing data from primary tumours and PDX models, we find that PDX establishment creates a genomic bottleneck, with 76% of PDX models being derived from a single primary tumour subclone. Despite this, multiple primary tumour subclones were capable of PDX establishment in regional PDX models, indicating that PDX libraries derived from multiple tumour regions can capture intratumour heterogeneity. Acquisition of somatic mutations continued during PDX model expansion, and was associated with APOBEC- or mismatch repair deficiency-induced mutational signatures in a subset of models. Overall, while NSCLC PDX models retain truncal genomic alterations, the absence of subclonal heterogeneity representative of the primary tumour is a major limitation. Our results emphasise the importance of characterising and monitoring intratumour heterogeneity in the context of pre-clinical cancer studies.
]]></description>
<dc:creator>Hynds, R. E.</dc:creator>
<dc:creator>Huebner, A.</dc:creator>
<dc:creator>Pearce, D. R.</dc:creator>
<dc:creator>Akarca, A. U.</dc:creator>
<dc:creator>Moore, D. A.</dc:creator>
<dc:creator>Ward, S.</dc:creator>
<dc:creator>Gowers, K. H. C.</dc:creator>
<dc:creator>Karasaki, T.</dc:creator>
<dc:creator>Hill, M. S.</dc:creator>
<dc:creator>Al Bakir, M.</dc:creator>
<dc:creator>Wilson, G. A.</dc:creator>
<dc:creator>Pich, O.</dc:creator>
<dc:creator>Sivakumar, M.</dc:creator>
<dc:creator>Ben Aissa, A.</dc:creator>
<dc:creator>Gronroos, E.</dc:creator>
<dc:creator>Chandrasekharan, D.</dc:creator>
<dc:creator>Kolluri, K. K.</dc:creator>
<dc:creator>Towns, R.</dc:creator>
<dc:creator>Wang, K.</dc:creator>
<dc:creator>Cook, D. E.</dc:creator>
<dc:creator>Bosshard-Carter, L.</dc:creator>
<dc:creator>Naceur-Lombardelli, C.</dc:creator>
<dc:creator>Rowan, A. J.</dc:creator>
<dc:creator>Veeriah, S.</dc:creator>
<dc:creator>Litchfield, K.</dc:creator>
<dc:creator>Quezada, S. A.</dc:creator>
<dc:creator>Janes, S. M.</dc:creator>
<dc:creator>Jamal-Hanjani, M.</dc:creator>
<dc:creator>Marafioti, T.</dc:creator>
<dc:creator>TRACERx Consortium,</dc:creator>
<dc:creator>McGranahan, N.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:date>2023-01-06</dc:date>
<dc:identifier>doi:10.1101/2023.01.06.521078</dc:identifier>
<dc:title><![CDATA[Genomic evolution of non-small cell lung cancer patient-derived xenograft models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.11.523553v1?rss=1">
<title>
<![CDATA[
Analysis of CDPK1 targets identifies a trafficking adaptor complex that regulates microneme exocytosis in Toxoplasma 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.11.523553v1?rss=1"
</link>
<description><![CDATA[
Apicomplexan parasites use Ca2+-regulated exocytosis to secrete essential virulence factors from specialized organelles called micronemes. Ca2+-dependent protein kinases (CDPKs) are required for microneme exocytosis; however, the molecular events that regulate trafficking and fusion of micronemes with the plasma membrane remain unresolved. Here, we combine sub-minute resolution phosphoproteomics and bio-orthogonal labeling of kinase substrates in Toxoplasma gondii to identify 163 proteins phosphorylated in a CDPK1-dependent manner. In addition to known regulators of secretion, we identify uncharacterized targets with predicted functions across signaling, gene expression, trafficking, metabolism, and ion homeostasis. One of the CDPK1 targets is a putative HOOK activating adaptor. In other eukaryotes, HOOK homologs form the FHF complex with FTS and FHIP to activate dynein-mediated trafficking of endosomes along microtubules. We show the FHF complex is partially conserved in T. gondii, consisting of HOOK, an FTS homolog, and two parasite-specific proteins (TGGT1_306920 and TGGT1_316650). CDPK1 kinase activity and HOOK are required for the rapid apical trafficking of micronemes as parasites initiate motility. Moreover, parasites lacking HOOK or FTS display impaired microneme protein secretion, leading to a block in the invasion of host cells. Taken together, our work provides a comprehensive catalog of CDPK1 targets and reveals how vesicular trafficking has been tuned to support a parasitic lifestyle.
]]></description>
<dc:creator>Chan, A. W.</dc:creator>
<dc:creator>Broncel, M.</dc:creator>
<dc:creator>Haseley, N. R.</dc:creator>
<dc:creator>Chakladar, S.</dc:creator>
<dc:creator>Andree, E.</dc:creator>
<dc:creator>Herneisen, A. L.</dc:creator>
<dc:creator>Shortt, E.</dc:creator>
<dc:creator>Treeck, M.</dc:creator>
<dc:creator>Lourido, S.</dc:creator>
<dc:date>2023-01-11</dc:date>
<dc:identifier>doi:10.1101/2023.01.11.523553</dc:identifier>
<dc:title><![CDATA[Analysis of CDPK1 targets identifies a trafficking adaptor complex that regulates microneme exocytosis in Toxoplasma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.10.523473v1?rss=1">
<title>
<![CDATA[
Femtosecond laser preparation of resin embedded samples for correlative microscopy workflows in life sciences 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.10.523473v1?rss=1"
</link>
<description><![CDATA[
Correlative multimodal imaging is a useful approach to investigate complex structural relations in life sciences across multiple scales. For these experiments, sample preparation workflows that are compatible with multiple imaging techniques must be established. In one such implementation, a fluorescently-labelled region of interest in a biological soft tissue sample can be imaged with light microscopy before staining the specimen with heavy metals, enabling follow-up higher resolution structural imaging at the targeted location, bringing context where it is required. Alternatively, or in addition to fluorescence imaging, other microscopy methods such as synchrotron X-ray computed tomography with propagation-based phase contrast (SXRT) or serial blockface scanning electron microscopy (SBF-SEM) might also be applied. When combining imaging techniques across scales, it is common that a volumetric region of interest (ROI) needs to be carved from the total sample volume before high resolution imaging with a subsequent technique can be performed. In these situations, the overall success of the correlative workflow depends on the precise targeting of the ROI and the trimming of the sample down to a suitable dimension and geometry for downstream imaging.

Here we showcase the utility of a novel femtosecond laser device to prepare microscopic samples (1) of an optimised geometry for synchrotron X-ray microscopy as well as (2) for subsequent volume electron microscopy applications, embedded in a wider correlative multimodal imaging workflow (Fig. 1).

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=172 SRC="FIGDIR/small/523473v1_fig1.gif" ALT="Figure 1">
View larger version (42K):
org.highwire.dtl.DTLVardef@1c72db6org.highwire.dtl.DTLVardef@18a234forg.highwire.dtl.DTLVardef@142bd0org.highwire.dtl.DTLVardef@4894f0_HPS_FORMAT_FIGEXP  M_FIG O_FLOATNOFigure 1.C_FLOATNO Correlative microscopy workflows incorporating samples processed using a femtosecond laser.

C_FIG
]]></description>
<dc:creator>Bosch, C.</dc:creator>
<dc:creator>Lindenau, J.</dc:creator>
<dc:creator>Pacureanu, A.</dc:creator>
<dc:creator>Peddie, C. J.</dc:creator>
<dc:creator>Majkut, M.</dc:creator>
<dc:creator>Douglas, A. C.</dc:creator>
<dc:creator>Carzaniga, R.</dc:creator>
<dc:creator>Rack, A.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:creator>Schaefer, A.</dc:creator>
<dc:creator>Stegmann, H.</dc:creator>
<dc:date>2023-01-12</dc:date>
<dc:identifier>doi:10.1101/2023.01.10.523473</dc:identifier>
<dc:title><![CDATA[Femtosecond laser preparation of resin embedded samples for correlative microscopy workflows in life sciences]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.12.523500v1?rss=1">
<title>
<![CDATA[
spinDrop: a droplet microfluidic platform to maximise single-cell sequencing information content 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.12.523500v1?rss=1"
</link>
<description><![CDATA[
Droplet microfluidic methods have massively increased the throughput of single-cell sequencing campaigns. The benefit of scale-up is, however, accompanied by increased background noise when processing challenging samples and the overall RNA capture efficiency is lower. These drawbacks stem from the lack of strategies to enrich for high-quality material or specific cell types at the moment of cell encapsulation and the absence of implementable multi-step enzymatic processes that increase capture. Here we alleviate both bottlenecks using fluorescence-activated droplet sorting to enrich for droplets that contain single viable cells, intact nuclei, fixed cells or target cell types and use reagent addition to droplets by picoinjection to perform multi-step lysis and reverse transcription. Our methodology increases gene detection rates fivefold, while reducing background noise by up to half. We harness these unique properties to deliver a high-quality molecular atlas of mouse brain development, despite starting with highly damaged input material, and provide an atlas of nascent RNA transcription during mouse organogenesis. Our method is broadly applicable to other droplet-based workflows to deliver sensitive and accurate single-cell profiling at a reduced cost.
]]></description>
<dc:creator>de Jonghe, J.</dc:creator>
<dc:creator>Kaminski, T. S.</dc:creator>
<dc:creator>Morse, D.</dc:creator>
<dc:creator>Tabaka, M.</dc:creator>
<dc:creator>Ellermann, A. L.</dc:creator>
<dc:creator>Kohler, T. N.</dc:creator>
<dc:creator>Amadei, G.</dc:creator>
<dc:creator>Handford, C.</dc:creator>
<dc:creator>Findlay, G.</dc:creator>
<dc:creator>Zernicka-Goetz, M.</dc:creator>
<dc:creator>Teichmann, S.</dc:creator>
<dc:creator>Hollfelder, F.</dc:creator>
<dc:date>2023-01-13</dc:date>
<dc:identifier>doi:10.1101/2023.01.12.523500</dc:identifier>
<dc:title><![CDATA[spinDrop: a droplet microfluidic platform to maximise single-cell sequencing information content]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.13.523922v1?rss=1">
<title>
<![CDATA[
Automated staging of zebrafish embryos with KimmelNet 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.13.523922v1?rss=1"
</link>
<description><![CDATA[
The zebrafish (Danio rerio), is an important biomedical model organism used in many disciplines. The phenomenon of developmental delay in zebrafish embryos has been widely reported as part of a mutant or treatment-induced phenotype. However, the detection and quantification of these delays is often achieved through manual observation with reference to staging guides, which is both time-consuming and subjective. We recently reported a machine learning-based classifier, capable of quantifying the developmental delay between two populations of zebrafish embryos. Here, we build on that work by introducing a deep learning-based model (KimmelNet) that has been trained to predict the age (hours post fertilisation) of populations of zebrafish embryos. We show that when KimmelNet is tested on 2D brightfield images of zebrafish embryos, the predictions generated agree closely with those expected from established approaches to staging. Random sampling of the test data demonstrate that KimmelNet can be used to detect developmental delay between two populations with high confidence based on as few as 100 images of each population. Finally, we show that KimmelNet generalises to previously unseen data, with limited transfer learning improving this performance significantly. With the ability to analyse tens of thousands of standard brightfield microscopy images on a timescale of minutes, we envisage that KimmelNet will be a valuable resource for the developmental biology community. Furthermore, the approach we have used could easily be adapted to generate models for other organisms.
]]></description>
<dc:creator>Barry, D. J.</dc:creator>
<dc:creator>Jones, R. A.</dc:creator>
<dc:creator>Renshaw, M. J.</dc:creator>
<dc:date>2023-01-14</dc:date>
<dc:identifier>doi:10.1101/2023.01.13.523922</dc:identifier>
<dc:title><![CDATA[Automated staging of zebrafish embryos with KimmelNet]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.15.524117v1?rss=1">
<title>
<![CDATA[
Biomimetic actin cortices shape cell-sized lipid vesicles 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.15.524117v1?rss=1"
</link>
<description><![CDATA[
Animal cells are shaped by a thin layer of actin filaments underneath the plasma membrane known as the actin cortex. This cortex stiffens the cell surface and thus opposes cellular deformation, yet also actively generates membrane protrusions by exerting polymerization forces. It is unclear how the interplay between these two opposing mechanical functions plays out to shape the cell surface. To answer this question, we reconstitute biomimetic actin cortices nucleated by the Arp2/3 complex inside cell-sized lipid vesicles. We show that thin Arp2/3-nucleated actin cortices strongly deform and rigidify the shapes of giant unilamellar vesicles and impart a shape memory on time scales that exceeds the time of actin turnover. In addition, actin cortices can produce finger-like membrane protrusions, showing that Arp2/3-mediated actin polymerization forces alone are sufficient to initiate protrusions in the absence of actin bundling or membrane curving proteins. Combining mathematical modeling and our experimental results reveals that the concentration of actin nucleating proteins, rather than actin polymerization speed, is crucial for protrusion formation. This is because locally concentrated actin polymerization forces can drive a positive feedback loop between recruitment of actin and its nucleators to drive membrane deformation. Our work paints a picture where the actin cortex can either drive or inhibit deformations depending on the local distribution of nucleators.

Significance StatementThe cells in our body must actively change shape in order to migrate, grow and divide, but they also need to maintain their shape to withstand external forces during tissue development. Cellular shape control results from an interplay between the plasma membrane and its underlying cortex, a shell composed of crosslinked actin filaments. Using cell-free reconstitution and mathematical modelling, we show that minimal biomimetic actin cortices can mechanically rigidify lipid vesicles while at the same time driving membrane protrusion formation. Our observations suggest that the spatial distribution of actin nucleation determines whether the actin cortex drives or inhibits membrane deformations.
]]></description>
<dc:creator>Baldauf, L.</dc:creator>
<dc:creator>Frey, F.</dc:creator>
<dc:creator>Perez, M. A.</dc:creator>
<dc:creator>Mladenov, M.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:creator>Idema, T.</dc:creator>
<dc:creator>Koenderink, G.</dc:creator>
<dc:date>2023-01-18</dc:date>
<dc:identifier>doi:10.1101/2023.01.15.524117</dc:identifier>
<dc:title><![CDATA[Biomimetic actin cortices shape cell-sized lipid vesicles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.19.524688v1?rss=1">
<title>
<![CDATA[
ARPC5 deficiency leads to severe early onset systemic inflammation and early mortality 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.19.524688v1?rss=1"
</link>
<description><![CDATA[
The seven subunit Arp2/3 complex drives the formation of branched actin networks that are essential for many cellular processes including cell migration. In humans, the ARPC5 subunit of the Arp2/3 complex is encoded by two paralogous genes (ARPC5 and ARPC5L), resulting in proteins with 67% identity. Through whole-exome sequencing, we identified a biallelic ARPC5 frameshift variant in a female child who presented with recurrent infections, multiple congenital anomalies, diarrhea, and thrombocytopenia, and suffered early demise from sepsis. Her consanguineous parents also had a previous child who died with similar clinical features. Using CRISPR/Cas9-mediated approaches, we demonstrate that loss of ARPC5 affects actin cytoskeleton organization and function, as well as chemokine-dependent cell migration in vitro. Homozygous Arpc5-/- mice do not survive past embryonic day 9 due to severe developmental defects, including loss of the second pharyngeal arch which contributes to craniofacial and heart development. Our results indicate that ARPC5 is important for both prenatal development and postnatal immune signaling, in a non-redundant manner with ARPC5L. Moreover, our observations add the ARPC5 locus to the list of genes that should be considered when patients present with syndromic early-onset immunodeficiency, particularly if recessive inheritance is suspected.
]]></description>
<dc:creator>Sindram, E.</dc:creator>
<dc:creator>Caballero-Oteyza, A.</dc:creator>
<dc:creator>Cogata, N.</dc:creator>
<dc:creator>Huang, S. C. M.</dc:creator>
<dc:creator>Alizadeh, Z.</dc:creator>
<dc:creator>Gamez-Diaz, L.</dc:creator>
<dc:creator>Fazlollahi, M. R.</dc:creator>
<dc:creator>Peng, X.</dc:creator>
<dc:creator>Grimbacher, B.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:creator>Proietti, M.</dc:creator>
<dc:date>2023-01-21</dc:date>
<dc:identifier>doi:10.1101/2023.01.19.524688</dc:identifier>
<dc:title><![CDATA[ARPC5 deficiency leads to severe early onset systemic inflammation and early mortality]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.23.525149v1?rss=1">
<title>
<![CDATA[
Mis-spliced transcripts generate de novo proteins in TDP-43-related ALS/FTD 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.23.525149v1?rss=1"
</link>
<description><![CDATA[
Functional loss of TDP-43, an RNA-binding protein genetically and pathologically linked to ALS and FTD, leads to inclusion of cryptic exons in hundreds of transcripts during disease. Cryptic exons can promote degradation of affected transcripts, deleteriously altering cellular function through loss-of-function mechanisms. However, the possibility of de novo protein synthesis from cryptic exon transcripts has not been explored. Here, we show that mRNA transcripts harboring cryptic exons generate de novo proteins both in TDP-43 deficient cellular models and in disease. Using coordinated transcriptomic and proteomic studies of TDP-43 depleted iPSC-derived neurons, we identified numerous peptides that mapped to cryptic exons. Cryptic exons identified in iPSC models were highly predictive of cryptic exons expressed in brains of patients with TDP-43 proteinopathy, including cryptic transcripts that generated de novo proteins. We discovered that inclusion of cryptic peptide sequences in proteins altered their interactions with other proteins, thereby likely altering their function. Finally, we showed that these de novo peptides were present in CSF from patients with ALS. The demonstration of cryptic exon translation suggests new mechanisms for ALS pathophysiology downstream of TDP-43 dysfunction and may provide a strategy for novel biomarker development.

One Sentence SummaryLoss of TDP-43 function results in the expression of de novo proteins from mis-spliced mRNA transcripts.
]]></description>
<dc:creator>Seddighi, S.</dc:creator>
<dc:creator>Qi, Y. A.</dc:creator>
<dc:creator>Brown, A.-L.</dc:creator>
<dc:creator>Wilkins, O. G.</dc:creator>
<dc:creator>Bereda, C.</dc:creator>
<dc:creator>Belair, C.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Prudencio, M.</dc:creator>
<dc:creator>Keuss, M.</dc:creator>
<dc:creator>Khandeshi, A.</dc:creator>
<dc:creator>Pickles, S.</dc:creator>
<dc:creator>Hill, S.</dc:creator>
<dc:creator>Hawrot, J.</dc:creator>
<dc:creator>Ramos, D.</dc:creator>
<dc:creator>Yuan, H.</dc:creator>
<dc:creator>Roberts, J.</dc:creator>
<dc:creator>Kelmer Sacramento, E.</dc:creator>
<dc:creator>Shah, S.</dc:creator>
<dc:creator>Nalls, M. A.</dc:creator>
<dc:creator>Colon-Mercado, J.</dc:creator>
<dc:creator>Reyes, J.</dc:creator>
<dc:creator>Ryan, V.</dc:creator>
<dc:creator>Nelson, M.</dc:creator>
<dc:creator>Cook, C.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Screven, L.</dc:creator>
<dc:creator>Kwan, J.</dc:creator>
<dc:creator>Shantaraman, A.</dc:creator>
<dc:creator>Ping, L.</dc:creator>
<dc:creator>Koike, Y.</dc:creator>
<dc:creator>Oskarsson, B.</dc:creator>
<dc:creator>Staff, N.</dc:creator>
<dc:creator>Duong, D.</dc:creator>
<dc:creator>Ahmed, A.</dc:creator>
<dc:creator>Secrier, M.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Jacobson, S.</dc:creator>
<dc:creator>Rohrer, J.</dc:creator>
<dc:creator>Malaspina, A.</dc:creator>
<dc:creator>Glass, J.</dc:creator>
<dc:creator>Ori, A.</dc:creator>
<dc:creator>Seyfried, N.</dc:creator>
<dc:creator>Maragkakis, M.</dc:creator>
<dc:creator>Petrucelli, L.</dc:creator>
<dc:creator>Fratta, P.</dc:creator>
<dc:creator>Ward, M. E.</dc:creator>
<dc:date>2023-01-23</dc:date>
<dc:identifier>doi:10.1101/2023.01.23.525149</dc:identifier>
<dc:title><![CDATA[Mis-spliced transcripts generate de novo proteins in TDP-43-related ALS/FTD]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.24.525325v1?rss=1">
<title>
<![CDATA[
Mechanistic insights into the interactions between cancer drivers and the tumour immune microenvironment 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.24.525325v1?rss=1"
</link>
<description><![CDATA[
The crosstalk between cancer and the tumour immune microenvironment (TIME) has attracted significant interest because of its impact on cancer evolution and response to treatment. Despite this, cancer-specific tumour-TIME interactions and their mechanisms of action are still poorly understood. Here we identified the interactions between cancer-specific genetic drivers and anti- or pro-tumour TIME features in individual samples of 32 cancer types. The resulting 477 TIME drivers are multifunctional genes whose alterations are selected early in cancer evolution and recur across and within cancer types. Moreover, the anti-tumour TIME driver burden is predictive of overall response to immunotherapy. Focusing on head and neck squamous cancer (HNSC), we rebuilt the functional networks linking specific TIME driver alterations to the TIME state. We showed that TIME driver alterations predict the immune profiles of HNSC molecular subtypes, and that deregulation of keratinization, apoptosis and interferon signalling underpin specific driver-TIME interactions. Overall, our study provides a comprehensive resource of TIME drivers giving mechanistic insights into their immune-regulatory role.
]]></description>
<dc:creator>Misetic, H.</dc:creator>
<dc:creator>Keddar, M. R.</dc:creator>
<dc:creator>Jeannon, J.-P.</dc:creator>
<dc:creator>Ciccarelli, F. D.</dc:creator>
<dc:date>2023-01-24</dc:date>
<dc:identifier>doi:10.1101/2023.01.24.525325</dc:identifier>
<dc:title><![CDATA[Mechanistic insights into the interactions between cancer drivers and the tumour immune microenvironment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.24.520089v1?rss=1">
<title>
<![CDATA[
Phenotyping of lymphoproliferative tumours generated in xenografts of non-small cell lung cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.24.520089v1?rss=1"
</link>
<description><![CDATA[
Patient-derived xenograft (PDX) models involve the engraftment of tumour tissue in immunocompromised mice and represent an important pre-clinixtcal oncology research. A limitation of non-small cell lung cancer (NSCLC) PDX model derivation in NOD-scid IL2Rgammanull (NSG) mice is that a subset of initial engraftments are of lymphocytic, rather than tumour origin. In the lung TRACERx PDX pipeline, lymphoproliferations occurred in 17.8% of lung adenocarcinoma and 10% of lung squamous cell carcinoma transplantations, despite none of these patients having a prior or subsequent clinical history of lymphoproliferative disease. Lymphoproliferations were predominantly human CD20+ B cells and had the immunophenotype expected for post-transplantation diffuse large B cell lymphoma. All lymphoproliferations expressed Epstein-Barr-encoded RNAs (EBER). Analysis of immunoglobulin light chain gene rearrangements in three tumours where multiple tumour regions had resulted in lymphoproliferations suggested that each had independent clonal origins. Overall, these data suggest the presence of B cell clones with lymphoproliferative potential within primary NSCLC tumours that are under continuous immune surveillance. Since these cells can be expanded following transplantation into NSG mice, our data highlight the value of quality control measures to identify lymphoproliferations within xenograft pipelines and support the incorporation of strategies to minimise lymphoproliferations during the early stages of xenograft establishment pipelines. To present the histology data herein, we developed a Python-based tool for generating patient-level pathology overview figures from whole-slide image files; PATHOverview is available on GitHub (https://github.com/EpiCENTR-Lab/PATHOverview).
]]></description>
<dc:creator>Pearce, D. R.</dc:creator>
<dc:creator>Akarca, A. U.</dc:creator>
<dc:creator>De Maeyer, R. P. H.</dc:creator>
<dc:creator>Kostina, E.</dc:creator>
<dc:creator>Huebner, A.</dc:creator>
<dc:creator>Sivakumar, M.</dc:creator>
<dc:creator>Karasaki, T.</dc:creator>
<dc:creator>Shah, K.</dc:creator>
<dc:creator>Janes, S. M.</dc:creator>
<dc:creator>McGranahan, N. E.</dc:creator>
<dc:creator>Reddy, V.</dc:creator>
<dc:creator>Akbar, A.</dc:creator>
<dc:creator>Moore, D. A.</dc:creator>
<dc:creator>Marafioti, T.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Hynds, R. E.</dc:creator>
<dc:date>2023-01-24</dc:date>
<dc:identifier>doi:10.1101/2023.01.24.520089</dc:identifier>
<dc:title><![CDATA[Phenotyping of lymphoproliferative tumours generated in xenografts of non-small cell lung cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.27.525863v1?rss=1">
<title>
<![CDATA[
Extracellular vesicles and co-isolated endogenous retroviruses differently affect dendritic cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.27.525863v1?rss=1"
</link>
<description><![CDATA[
Cells secrete membrane-enclosed extracellular vesicles (EVs) and non-vesicular nanoparticles (ENPs) that may play a role in intercellular communication. Tumor-derived EVs have been proposed either to induce immune priming of antigen presenting cells, or, to be immuno-suppressive agents promoting tumor immune escape. We suspect that such disparate functions are due to variable composition in EV subtypes and ENPs of the analyzed EV preparations. We aimed to exhaustively characterize the array of secreted EVs and ENPs of murine tumor cell lines. Unexpectedly, we identified virus-like particles (VLPs) from endogenous murine leukemia virus in preparations of EVs produced by tumor cells. We established a robust protocol to separate small (s)EVs from VLPs and ENPs. We compared their protein composition and analyzed their functional interaction with target dendritic cells (DCs). ENPs were poorly captured and did not affect DCs. sEVs specifically induced DC death. A mixed EV/VLP preparation was the most efficient to induce DC maturation and antigen presentation. Our results call for systematic re-evaluation of the respective proportions and functions of non-viral EVs and VLPs produced by tumors and their contribution to anti-tumor immune responses and to tumor progression.
]]></description>
<dc:creator>Cocozza, F.</dc:creator>
<dc:creator>martin jaular, l.</dc:creator>
<dc:creator>lippens, l.</dc:creator>
<dc:creator>di cicco, a.</dc:creator>
<dc:creator>arribas, y. a.</dc:creator>
<dc:creator>dingli, f.</dc:creator>
<dc:creator>richard, m.</dc:creator>
<dc:creator>merle, l.</dc:creator>
<dc:creator>Poullet, P.</dc:creator>
<dc:creator>loew, d.</dc:creator>
<dc:creator>Hendrix, A.</dc:creator>
<dc:creator>Kassiotis, G.</dc:creator>
<dc:creator>joliot, a.</dc:creator>
<dc:creator>tkach, m.</dc:creator>
<dc:creator>Thery, C.</dc:creator>
<dc:date>2023-01-28</dc:date>
<dc:identifier>doi:10.1101/2023.01.27.525863</dc:identifier>
<dc:title><![CDATA[Extracellular vesicles and co-isolated endogenous retroviruses differently affect dendritic cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.29.526106v1?rss=1">
<title>
<![CDATA[
Plasmodium ARK2-EB1 axis drives the unconventional spindle dynamics, scaffold formation, and chromosome segregation of sexual transmission stages 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.29.526106v1?rss=1"
</link>
<description><![CDATA[
Mechanisms of cell division are remarkably diverse, suggesting the underlying molecular networks among eukaryotes differ extensively. The Aurora family of kinases orchestrates the process of chromosome segregation and cytokinesis during cell division through precise spatiotemporal regulation of their catalytic activities by distinct scaffolds. Plasmodium spp., the causative agents of malaria, are unicellular eukaryotes that have three divergent aurora-related kinases (ARKs) and lack most canonical scaffolds/activators. The parasite uses unconventional modes of chromosome segregation during endomitosis and meiosis in sexual transmission stages within mosquito host. This includes a rapid threefold genome replication from 1N to 8N with successive cycles of closed mitosis, spindle formation and chromosome segregation within eight minutes (termed male gametogony). Kinome studies had previously suggested likely essential functions for all three Plasmodium ARKs during asexual mitotic cycles; however, little is known about their location, function, or their scaffolding molecules during unconventional sexual proliferative stages. Using a combination of super-resolution microscopy, mass spectrometry, and live-cell fluorescence imaging, we set out to investigate the role of the atypical Aurora paralog ARK2 to proliferative sexual stages using rodent malaria model Plasmodium berghei. We find that ARK2 primarily localises to the spindle apparatus in the vicinity of kinetochores during both mitosis and meiosis. Interactomics and co-localisation studies reveal a unique ARK2 scaffold at the spindle including the microtubule plus end-binding protein EB1, lacking conserved Aurora scaffold proteins. Gene function studies indicate complementary functions of ARK2 and EB1 in driving endomitotic divisions and thereby parasite transmission. Our discovery of a novel Aurora kinase spindle scaffold underlines the emerging flexibility of molecular networks to rewire and drive unconventional mechanisms of chromosome segregation in the malaria parasite Plasmodium.
]]></description>
<dc:creator>Zeeshan, M.</dc:creator>
<dc:creator>Rea, E.</dc:creator>
<dc:creator>Abel, S.</dc:creator>
<dc:creator>Vukusic, K.</dc:creator>
<dc:creator>Markus, R.</dc:creator>
<dc:creator>Brady, D.</dc:creator>
<dc:creator>Eze, A. A.</dc:creator>
<dc:creator>Rashpa, R.</dc:creator>
<dc:creator>Balestra, A.</dc:creator>
<dc:creator>Bottrill, A.</dc:creator>
<dc:creator>Brochet, M.</dc:creator>
<dc:creator>Guttery, D.</dc:creator>
<dc:creator>Tolic, I. M.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:creator>Le Roch, K. G.</dc:creator>
<dc:creator>Tromer, E.</dc:creator>
<dc:creator>Tewari, R.</dc:creator>
<dc:date>2023-01-31</dc:date>
<dc:identifier>doi:10.1101/2023.01.29.526106</dc:identifier>
<dc:title><![CDATA[Plasmodium ARK2-EB1 axis drives the unconventional spindle dynamics, scaffold formation, and chromosome segregation of sexual transmission stages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.31.526399v1?rss=1">
<title>
<![CDATA[
The C9Orf72 hexanucleotide repeat expansion transcript forms insoluble aggregates facilitated by multimolecular G-quadruplex structures 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.31.526399v1?rss=1"
</link>
<description><![CDATA[
Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are neurodegenerative diseases that exist on a clinico-pathogenetic spectrum, designated ALS/FTD. The most common genetic cause of ALS/FTD is the expansion of the intronic hexanucleotide repeat (GGGGCC)nin C9orf72. Here, we investigated the formation of nucleic-acid secondary structures in these expansion repeats, and their role in generating condensates characteristic of the diseases. We observed significant aggregation of the hexanucleotide sequence (GGGGCC)n, which we associated to the formation of multimolecular G-quadruplexes (mG4s), using a range of biophysical techniques. Exposing the condensates to G4-unfolding conditions led to prompt disassembly, highlighting the key role of mG4-formation in the condensation process. We further validated the biological relevance of our findings by demonstrating the ability of a G4-selective fluorescent probe to penetrate C9orf72 mutant human motor neurons derived from ALS patients, which revealed clear fluorescent signal in putative condensates. Our findings strongly suggest that RNA G- rich repetitive sequences can form protein-free condensates sustained by multimolecular G- quadruplexes, highlighting their potential relevance as therapeutic targets for C9orf72 mutation related ALS and FTD.



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]]></description>
<dc:creator>Raguseo, F.</dc:creator>
<dc:creator>Tanase, D.</dc:creator>
<dc:creator>Malouf, L.</dc:creator>
<dc:creator>Rubio Sanchez, R.</dc:creator>
<dc:creator>Elani, Y.</dc:creator>
<dc:creator>Di Michele, L.</dc:creator>
<dc:creator>Di Antonio, M.</dc:creator>
<dc:date>2023-01-31</dc:date>
<dc:identifier>doi:10.1101/2023.01.31.526399</dc:identifier>
<dc:title><![CDATA[The C9Orf72 hexanucleotide repeat expansion transcript forms insoluble aggregates facilitated by multimolecular G-quadruplex structures]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.01.526741v1?rss=1">
<title>
<![CDATA[
Imprinted anti-hemagglutinin and anti-neuraminidase antibody responses after childhood infections of A(H1N1) and A(H1N1)pdm09 influenza viruses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.01.526741v1?rss=1"
</link>
<description><![CDATA[
Immune imprinting is a driver known to shape the anti-hemagglutinin (HA) antibody landscape of individuals born within the same birth cohort. With the HA and neuraminidase (NA) proteins evolving at different rates under immune selection pressures, anti-HA and anti-NA antibody responses since childhood influenza infections have not been evaluated in parallel at the individual level. This is partly due to the limited knowledge of changes in NA antigenicity, as seasonal influenza vaccines have focused on generating neutralising anti-HA antibodies against HA antigenic variants. Here we systematically characterised the NA antigenic variants of seasonal A(H1N1) viruses from 1977 to 1991 and completed the antigenic profile of N1 NAs from 1977 to 2015. We identified that NA proteins of A/USSR/90/77, A/Singapore/06/86, and A/Texas/36/91 were antigenically distinct and mapped N386K as a key determinant of the NA antigenic change from A/USSR/90/77 to A/Singapore/06/86. With comprehensive panels of HA and NA antigenic variants of A(H1N1) and A(H1N1)pdm09 viruses, we determined hemagglutinin inhibition (HI) and neuraminidase inhibition (NI) antibodies from 130 subjects born between 1950-2015. Age-dependent imprinting was observed for both anti-HA and anti-NA antibodies, with the peak HI and NI titers predominantly detected from subjects at 4-12 years old during the year of initial virus isolation, except the age-independent anti-HA antibody response against A(H1N1)pdm09 viruses. More participants possessed antibodies that reacted to multiple antigenically distinct NA proteins than those with antibodies that reacted to multiple antigenically distinct HA proteins. Our results support the need to include NA proteins in seasonal influenza vaccine preparations.

IMPORTANCESeasonal influenza vaccines have aimed to generate neutralizing anti-HA antibodies for protection since licensure. More recently, anti-NA antibodies have been established as an additional correlate of protection. While HA and NA antigenic changes occurred discordantly, the anti-HA and anti-NA antibody profiles have rarely been analysed in parallel at the individual level, due to the limited knowledge on NA antigenic changes. By characterizing NA antigenic changes of A(H1N1) viruses, we determined the anti-HA and anti-NA antibody landscape against antigenically distinct A(H1N1) and A(H1N1)pdm09 viruses using sera of 130 subjects born between 1950-2015. We observed age-dependent imprinting of both anti-HA and anti-NA antibodies against strains circulated during the first decade of life. 67.7% (88/130) and 90% (117/130) of participants developed cross-reactive antibodies to multiple HA and NA antigens at titers [&ge;]1:40. With slower NA antigenic changes and cross-reactive anti-NA antibody responses, including NA protein in influenza vaccine preparation may enhance vaccine efficacy. (150 words)
]]></description>
<dc:creator>Daulagala, P.</dc:creator>
<dc:creator>Mann, B.</dc:creator>
<dc:creator>Leung, K.</dc:creator>
<dc:creator>Lau, E.</dc:creator>
<dc:creator>Yung, L.</dc:creator>
<dc:creator>Lei, R.</dc:creator>
<dc:creator>Nizami, S. I. N.</dc:creator>
<dc:creator>Wu, J. T.</dc:creator>
<dc:creator>Chiu, S. S.</dc:creator>
<dc:creator>Daniels, R. S.</dc:creator>
<dc:creator>Wu, N. C.</dc:creator>
<dc:creator>Wentworth, D. E.</dc:creator>
<dc:creator>Peiris, J. S. M.</dc:creator>
<dc:creator>Yen, H.-L.</dc:creator>
<dc:date>2023-02-02</dc:date>
<dc:identifier>doi:10.1101/2023.02.01.526741</dc:identifier>
<dc:title><![CDATA[Imprinted anti-hemagglutinin and anti-neuraminidase antibody responses after childhood infections of A(H1N1) and A(H1N1)pdm09 influenza viruses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.02.526785v1?rss=1">
<title>
<![CDATA[
PerTurboID: A targeted in situ method to measure changes in a local protein environment reveals the impact of kinase deletion on cytoadhesion in malaria causing parasites 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.02.526785v1?rss=1"
</link>
<description><![CDATA[
Reverse genetics is key to understanding protein function, but the mechanistic connection between a gene of interest and the observed phenotype is not always clear. Here we describe the use of proximity labeling using TurboID and site-specific quantification of biotinylated peptides to measure changes to the local protein environment of selected targets upon perturbation. We apply this technique, which we call PerTurboID, to understand how the P. falciparum exported kinase, FIKK4.1, regulates the function of the major virulence factor of the malaria causing parasite, PfEMP1. We generated independent TurboID fusions of 2 proteins that are predicted substrates of FIKK4.1 in a FIKK4.1 conditional KO parasite line. Comparing the abundance of site-specific biotinylated peptides between wildtype and kinase deletion lines reveals the differential accessibility of proteins to biotinylation, indicating changes to localization, protein-protein interactions, or protein structure which are mediated by FIKK4.1 activity. We further show that FIKK4.1 is likely the only FIKK kinase that controls surface levels of PfEMP1, but not other surface antigens, on the infected red blood cell under standard culture conditions. We believe PerTurboID is broadly applicable to study the impact of genetic or environmental perturbation on a selected cellular niche.
]]></description>
<dc:creator>Davies, H.</dc:creator>
<dc:creator>Belda, H.</dc:creator>
<dc:creator>Broncel, M.</dc:creator>
<dc:creator>Dalimot, J.</dc:creator>
<dc:creator>Treeck, M.</dc:creator>
<dc:date>2023-02-03</dc:date>
<dc:identifier>doi:10.1101/2023.02.02.526785</dc:identifier>
<dc:title><![CDATA[PerTurboID: A targeted in situ method to measure changes in a local protein environment reveals the impact of kinase deletion on cytoadhesion in malaria causing parasites]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.01.526632v1?rss=1">
<title>
<![CDATA[
A chemoproteomic platform for reactive fragment profiling against the deubiquitinases 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.01.526632v1?rss=1"
</link>
<description><![CDATA[
Chemoproteomics is a powerful method capable of detecting interactions between small molecules and the proteome, however its use as a high-throughput screening method for chemical libraries has so far been limited. To address this need, we have further developed a chemoproteomics workflow to screen cysteine reactive covalent fragments in cell lysates against the deubiquitinating (DUB) enzymes using activity-based protein profiling. By using targeted ubiquitin probes, we have addressed sensitivity and affinity limitations, enabling target identification and covalent fragment library profiling in a 96-well plate format. The use of data independent acquisition (DIA) methods for MS analysis combined with automated Evosep liquid chromatography (LC) reduced instrument runtimes to 21 minutes per sample and simplified the workflow. In this proof-of-concept study, we have profiled 138 covalent fragments against 57 DUB proteins and validated four hit fragments against OTUD7B and UCHL3 through site identification experiments and orthogonal biochemical activity assays.

Graphical abstract

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]]></description>
<dc:creator>Cookson, R.</dc:creator>
<dc:creator>Vuorinen, A.</dc:creator>
<dc:creator>Pettinger, J.</dc:creator>
<dc:creator>Kennedy, C. R.</dc:creator>
<dc:creator>Kirkpatrick, J. M.</dc:creator>
<dc:creator>Peltier-Heap, R. E.</dc:creator>
<dc:creator>Powell, A.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Skehel, M.</dc:creator>
<dc:creator>House, D.</dc:creator>
<dc:creator>Rittinger, K.</dc:creator>
<dc:creator>Bush, J. T.</dc:creator>
<dc:date>2023-02-03</dc:date>
<dc:identifier>doi:10.1101/2023.02.01.526632</dc:identifier>
<dc:title><![CDATA[A chemoproteomic platform for reactive fragment profiling against the deubiquitinases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.03.527081v1?rss=1">
<title>
<![CDATA[
MICA: A multi-omics method to predict gene regulatory networks in early human embryos 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.03.527081v1?rss=1"
</link>
<description><![CDATA[
Recent advances in single-cell-omics have been transformative to characterise cell types in challenging to study biological contexts. It is technically difficult to infer transcription factor-gene regulatory interactions from these datasets, especially in contexts with limited single-cell sample number such as the early human embryo. Here we systematically assessed the application of four different linear or non-linear gene regulatory network (GRN) prediction strategies to single-cell simulated and human embryo transcriptome datasets. We also compared how gene expression normalisation methods impact on regulatory network predictions, finding that TPM (transcripts per million reads) outperformed alternative methods. We identified more reproducible GRN inferences using a non-linear method based on mutual information (MI) applied to single-cell transcriptome datasets refined with chromatin accessibility (CA) (called MICA), compared to alternative network prediction methods tested. MICA captures complex non-monotonic dependencies and feedback loops. Using MICA, we generated the first GRN inferences in early human development. MICA predicted co-localisation of the AP-1 transcription factor subunit proto-oncogene JUND and the TFAP2C transcription factor AP-2{psi} in human preimplantation embryos. These interactions were supported by previous findings in other developmental and stem cell contexts. Overall, our comparative analysis of gene regulatory network prediction methods defines a pipeline that can be applied to single-cell multi-omics datasets in especially challenging contexts to infer interactions between transcription factor expression and target gene regulation.
]]></description>
<dc:creator>Alanis-Lobato, G.</dc:creator>
<dc:creator>Bartlett, T. E.</dc:creator>
<dc:creator>Huang, Q.</dc:creator>
<dc:creator>Simon, C.</dc:creator>
<dc:creator>McCarthy, A.</dc:creator>
<dc:creator>Elder, K.</dc:creator>
<dc:creator>Snell, P.</dc:creator>
<dc:creator>Christie, L.</dc:creator>
<dc:creator>Niakan, K.</dc:creator>
<dc:date>2023-02-03</dc:date>
<dc:identifier>doi:10.1101/2023.02.03.527081</dc:identifier>
<dc:title><![CDATA[MICA: A multi-omics method to predict gene regulatory networks in early human embryos]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.09.527935v1?rss=1">
<title>
<![CDATA[
A Potent and Selective CDKL5/GSK3 Chemical Probe is Neuroprotective 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.09.527935v1?rss=1"
</link>
<description><![CDATA[
Despite mediating several essential processes in the brain, including during development, cyclin-dependent kinase-like 5 (CDKL5) remains a poorly characterized human protein kinase. Accordingly, its substrates, functions, and regulatory mechanisms have not been fully described. We realized that availability of a potent and selective small molecule probe targeting CDKL5 could enable illumination of its roles in normal development as well as in diseases where it has become aberrant due to mutation. We prepared analogs of AT-7519, a known inhibitor of several cyclin dependent and cyclin-dependent kinase-like kinases that has been advanced into Phase II clinical trials. We identified analog 2 as a highly potent and cell-active chemical probe for CDKL5/GSK3 (glycogen synthase kinase 3). Evaluation of its kinome-wide selectivity confirmed that analog 2 demonstrates excellent selectivity and only retains GSK3/{beta} affinity. As confirmation that our chemical probe is a high-quality tool to use in directed biological studies, we demonstrated inhibition of downstream CDKL5 and GSK3/{beta} signaling and solved a co-crystal structure of analog 2 bound to CDKL5. A structurally similar analog (4) proved to lack CDKL5 affinity and maintain potent and selective inhibition of GSK3/{beta}. Finally, we used our chemical probe pair (2 and 4) to demonstrate that inhibition of CDKL5 and/or GSK3/{beta} promotes the survival of human motor neurons exposed to endoplasmic reticulum (ER) stress. We have demonstrated a neuroprotective phenotype elicited by our chemical probe pair and exemplified the utility of our compounds to characterize the role of CDKL5/GSK3 in neurons and beyond.
]]></description>
<dc:creator>Ong, H. W.</dc:creator>
<dc:creator>Liang, Y.</dc:creator>
<dc:creator>Richardson, W.</dc:creator>
<dc:creator>Lowry, E. R.</dc:creator>
<dc:creator>Wells, C. I.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Silvestre, M.</dc:creator>
<dc:creator>Dempster, K.</dc:creator>
<dc:creator>Silvaroli, J. A.</dc:creator>
<dc:creator>Smith, J. L.</dc:creator>
<dc:creator>Wichterle, H.</dc:creator>
<dc:creator>Pabla, N. S.</dc:creator>
<dc:creator>Ultanir, S. K.</dc:creator>
<dc:creator>Bullock, A. N.</dc:creator>
<dc:creator>Drewry, D.</dc:creator>
<dc:creator>Axtman, A. D.</dc:creator>
<dc:date>2023-02-10</dc:date>
<dc:identifier>doi:10.1101/2023.02.09.527935</dc:identifier>
<dc:title><![CDATA[A Potent and Selective CDKL5/GSK3 Chemical Probe is Neuroprotective]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.08.527637v1?rss=1">
<title>
<![CDATA[
Palmitoyl transferase ZDHHC20 promotes pancreatic cancer metastasis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.08.527637v1?rss=1"
</link>
<description><![CDATA[
Metastasis is one of the defining features of pancreatic ductal adenocarcinoma (PDAC) that contributes to poor prognosis. In this study, the palmitoyl transferase ZDHHC20 was identified in an in vivo shRNA screen as critical for metastatic outgrowth, with no effect on proliferation and migration in vitro, or primary PDAC growth in mice. This phenotype is abrogated in immunocompromised animals, and in animals with depleted natural killer (NK) cells, indicating that ZDHHC20 affects the interaction of tumour cells and the innate immune system. Using a chemical genetics platform for ZDHHC20-specific substrate profiling, a number of novel substrates of this enzyme were identified. These results describe a role for palmitoylation in enabling distant metastasis that could not have been detected using in vitro screening approaches and identify potential effectors through which ZDHHC20 promotes metastasis of PDAC.
]]></description>
<dc:creator>Tomic, G.</dc:creator>
<dc:creator>Sheridan, C.</dc:creator>
<dc:creator>Refermat, A.</dc:creator>
<dc:creator>Baggelaar, M.</dc:creator>
<dc:creator>Sipthorp, J.</dc:creator>
<dc:creator>Sudarshan, B.</dc:creator>
<dc:creator>Ocasio, C. A.</dc:creator>
<dc:creator>Suarez-Bonnet, A.</dc:creator>
<dc:creator>Priestnall, S.</dc:creator>
<dc:creator>Herbert, E.</dc:creator>
<dc:creator>Tate, E.</dc:creator>
<dc:creator>Downward, J.</dc:creator>
<dc:date>2023-02-08</dc:date>
<dc:identifier>doi:10.1101/2023.02.08.527637</dc:identifier>
<dc:title><![CDATA[Palmitoyl transferase ZDHHC20 promotes pancreatic cancer metastasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.09.527483v1?rss=1">
<title>
<![CDATA[
Enzyme-less nanopore detection of post-translational modifications within long polypeptides 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.09.527483v1?rss=1"
</link>
<description><![CDATA[
Means to sequence DNA and RNA quickly and cheaply have revolutionized biology and medicine. The ability to analyse cellular proteins and their millions of variants would be an advance of comparable importance, but requires a fresh technical approach. We use electroosmosis for the non-enzymatic capture, unfolding and translocation of individual polypeptides of more than 1200 residues by a protein nanopore. By monitoring the ionic current carried by the nanopore, we locate post-translational modifications deep within the polypeptide chains, and thereby lay the groundwork for obtaining inventories of the proteoforms in cells and tissues.
]]></description>
<dc:creator>Martin-Baniandres, P.</dc:creator>
<dc:creator>Lan, W.-H.</dc:creator>
<dc:creator>Board, S.</dc:creator>
<dc:creator>Romero-ruiz, M.</dc:creator>
<dc:creator>Garcia-Manyes, S.</dc:creator>
<dc:creator>Qing, Y.</dc:creator>
<dc:creator>Bayley, H.</dc:creator>
<dc:date>2023-02-09</dc:date>
<dc:identifier>doi:10.1101/2023.02.09.527483</dc:identifier>
<dc:title><![CDATA[Enzyme-less nanopore detection of post-translational modifications within long polypeptides]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.10.528004v1?rss=1">
<title>
<![CDATA[
The yeast RNA methylation complex consists of conserved yet reconfigured components with m6A-dependent and independent roles 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.10.528004v1?rss=1"
</link>
<description><![CDATA[
N6-methyladenosine (m6A), the most abundant mRNA modification, is deposited in mammals/insects/plants by m6A methyltransferase complexes (MTC) comprising a catalytic subunit and at least five additional proteins. The yeast MTC is critical for meiosis and was known to comprise three proteins, of which two were conserved. We uncover three novel MTC components (Kar4/Ygl036w-Vir1/Dyn2). All MTC subunits, except for Dyn2, are essential for m6A deposition and have corresponding mammalian MTC orthologs. Unlike the mammalian bipartite MTC, the yeast MTC is unipartite, yet multifunctional. The mRNA interacting module, comprising Ime4, Mum2, Vir1, and Kar4, exerts the MTCs m6A-independent function, while Slz1 enables the MTC catalytic function in m6A deposition. Both functions are critical for meiotic progression. Kar4 also has a mechanistically separate role from the MTC during mating. The yeast MTC constituents play distinguishable m6A-dependent, MTC-dependent and MTC-independent functions, highlighting their complexity and paving the path towards dissecting multi-layered MTC functions in mammals.
]]></description>
<dc:creator>Ensinck, I.</dc:creator>
<dc:creator>Maman, A.</dc:creator>
<dc:creator>Albihlal, W.</dc:creator>
<dc:creator>Lassandro, M.</dc:creator>
<dc:creator>Salzano, G.</dc:creator>
<dc:creator>Sideri, T.</dc:creator>
<dc:creator>Howell, S.</dc:creator>
<dc:creator>Calvani, E.</dc:creator>
<dc:creator>Patel, H.</dc:creator>
<dc:creator>Bushkin, G. G.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:creator>Skehel, M.</dc:creator>
<dc:creator>Casanal, A.</dc:creator>
<dc:creator>Schwartz, S.</dc:creator>
<dc:creator>van Werven, F.</dc:creator>
<dc:date>2023-02-10</dc:date>
<dc:identifier>doi:10.1101/2023.02.10.528004</dc:identifier>
<dc:title><![CDATA[The yeast RNA methylation complex consists of conserved yet reconfigured components with m6A-dependent and independent roles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.10.527985v1?rss=1">
<title>
<![CDATA[
The ecdysone receptor promotes or suppresses proliferation according toligand level 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.10.527985v1?rss=1"
</link>
<description><![CDATA[
Steroid hormones control various cellular activities in a context-dependent manner. For example, ecdysone, which acts through a type II nuclear receptor, has seemingly opposite effects in Drosophila wing precursors, promoting proliferation during larval stages, and triggering proliferation arrest at pupariation. We find that wing precursors proliferate normally in the complete absence of the ecdysone receptor (EcR), whether ecdysone is present or not, suggesting that ecdysone overrides a default antiproliferative activity of the receptor. By contrast, termination of proliferation by high concentration of 20E at the end of larval life involves conventional gene regulation by the ligand-receptor complex. The switch from one mode of regulation to the other is determined by ligand level, as measured with a calibrated EcR transcriptional reporter and ex vivo proliferation assays. Accordingly, RNA Seq analysis uncovers distinct transcriptional responses to different doses of ecdysone. Some genes are only activated at high doses (high threshold targets) and likely to comprise genes that stop proliferation at pupariation, when ecdysone titres are high. We find that other target genes respond to all physiological concentrations of ecdysone. Some of these genes are known to promote proliferation and could therefore contribute to the pro-proliferation activity of low-level ecdysone. Finally, we show mathematically and with synthetic reporters that relatively simple combinations of regulatory elements can recapitulate the behaviour of both types of target genes.
]]></description>
<dc:creator>Perez-Mockus, G.</dc:creator>
<dc:creator>Cocconi, L.</dc:creator>
<dc:creator>Alexandre, C.</dc:creator>
<dc:creator>Aerne, B. L.</dc:creator>
<dc:creator>Salbreux, G.</dc:creator>
<dc:creator>Vincent, J.-P.</dc:creator>
<dc:date>2023-02-11</dc:date>
<dc:identifier>doi:10.1101/2023.02.10.527985</dc:identifier>
<dc:title><![CDATA[The ecdysone receptor promotes or suppresses proliferation according toligand level]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.11.528101v1?rss=1">
<title>
<![CDATA[
Unique Amphipathic α-helix Drives Membrane Insertion and Enzymatic Activity of ATG3 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.11.528101v1?rss=1"
</link>
<description><![CDATA[
AbstractAutophagosome biogenesis requires a localized perturbation of lipid membrane dynamics and a unique protein-lipid conjugate. Autophagy-related (ATG) proteins catalyze this biogenesis on cellular membranes, but the underlying molecular mechanism remains unclear. Focusing on the final step of the protein-lipid conjugation reaction, ATG8/LC3 lipidation, we show how membrane association of the conjugation machinery is organized and fine-tuned at the atomistic level. Amphipathic -helices in ATG3 proteins (AHATG3) are found to have low hydrophobicity and to be less bulky. Molecular dynamics simulations reveal that AHATG3 regulates the dynamics and accessibility of the thioester bond of the ATG3[~]LC3 conjugate to lipids, allowing covalent lipidation of LC3. Live cell imaging shows that the transient membrane association of ATG3 with autophagic membranes is governed by the less bulky- hydrophobic feature of AHATG3. Collectively, the unique properties of AHATG3 facilitate protein- lipid bilayer association leading to the remodeling of the lipid bilayer required for the formation of autophagosomes.

TeaserWe uncover the unique biophysical property of amphipathic -helix essential for autophagy
]]></description>
<dc:creator>Nishimura, T.</dc:creator>
<dc:creator>Lazzeri, G.</dc:creator>
<dc:creator>Mizushima, N.</dc:creator>
<dc:creator>Covino, R.</dc:creator>
<dc:creator>Tooze, S.</dc:creator>
<dc:date>2023-02-12</dc:date>
<dc:identifier>doi:10.1101/2023.02.11.528101</dc:identifier>
<dc:title><![CDATA[Unique Amphipathic α-helix Drives Membrane Insertion and Enzymatic Activity of ATG3]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.10.527983v1?rss=1">
<title>
<![CDATA[
Stabilisation of β-Catenin-WNT signalling by USP10 in APC-truncated colorectal cancer drives cancer stemness and enables super-competitor signalling 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.10.527983v1?rss=1"
</link>
<description><![CDATA[
The contribution of deubiquitylating enzymes to {beta}-Catenin stabilisation in intestinal stem cells and colorectal cancer (CRC) is poorly understood. Here, we report the deubiquitylase USP10 as an APC-truncation- specific enhancer of {beta}-Catenin stability, potentiating WNT signalling in CRC and cancer stem cells. Mechanistically, interaction studies in various CRC cell lines and in vitro binding studies, together with computational modelling, revealed that USP10 binding to {beta}-Catenin is mediated via the unstructured N-terminus of USP10 and requires the absence of full-length APC. Notably, loss of USP10 in CRISPR engineered intestinal organoids reduces tumorigenic properties of CRC and blocks the super competitor-signalling of APC-mutated CRC. Furthermore, reduction of USP10 induces the expression of differentiation genes, and opposes the APC-truncated phenotype in an intestinal hyperplasia model of D.melanogaster.

Taken together, our findings reveal USP10s role in intestinal tumourigenesis by stabilising {beta}-Catenin, leading to aberrant WNT signalling, enhancing cancer cell stemness and implicate the DUB USP10 as a cancer specific therapeutic vulnerability in Apc truncated CRC.
]]></description>
<dc:creator>Reissland, M.</dc:creator>
<dc:creator>Hartmann, O.</dc:creator>
<dc:creator>Tauch, S.</dc:creator>
<dc:creator>Prieto-Garcia, C.</dc:creator>
<dc:creator>Schulte, C.</dc:creator>
<dc:creator>Solvei, D.</dc:creator>
<dc:creator>Loebbert, S.</dc:creator>
<dc:creator>Jacomin, A. C.</dc:creator>
<dc:creator>Pesic, M.</dc:creator>
<dc:creator>Bugter, J. M.</dc:creator>
<dc:creator>Schuelein-Voelk, C.</dc:creator>
<dc:creator>Fuss, C.</dc:creator>
<dc:creator>Pahor, N.</dc:creator>
<dc:creator>Ade, C.</dc:creator>
<dc:creator>Buck, V.</dc:creator>
<dc:creator>Potente, M.</dc:creator>
<dc:creator>Li, V.</dc:creator>
<dc:creator>Beliu, G.</dc:creator>
<dc:creator>Wiegering, A.</dc:creator>
<dc:creator>Bitman-Lotan, E.</dc:creator>
<dc:creator>Grossmann, T.</dc:creator>
<dc:creator>Rosenfeldt, M.</dc:creator>
<dc:creator>Eilers, M.</dc:creator>
<dc:creator>Maric, H.</dc:creator>
<dc:creator>Maurice, M.</dc:creator>
<dc:creator>Greten, F.</dc:creator>
<dc:creator>Dikic, I.</dc:creator>
<dc:creator>Orian, A.</dc:creator>
<dc:creator>Gallant, P.</dc:creator>
<dc:creator>Diefenbacher, M. E.</dc:creator>
<dc:date>2023-02-11</dc:date>
<dc:identifier>doi:10.1101/2023.02.10.527983</dc:identifier>
<dc:title><![CDATA[Stabilisation of β-Catenin-WNT signalling by USP10 in APC-truncated colorectal cancer drives cancer stemness and enables super-competitor signalling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.17.528960v1?rss=1">
<title>
<![CDATA[
Human Integrator provides a quality checkpoint during elongation to facilitate RNA polymerase II processivity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.17.528960v1?rss=1"
</link>
<description><![CDATA[
Integrator is a multi-subunit complex that directly interacts with the C-terminal domain (CTD) of RNA polymerase II (RNAPII). Through its RNA endonuclease activity, Integrator is required for 3'-end processing of both non-coding and coding transcripts. Here we demonstrate that depleting Integrator subunit 11 (INTS11), the main catalytic subunit of the Integrator complex, leads to a global elongation defect as a result of decreased polymerase processivity. We observe this defect in the region approximately 12 to 35 kb downstream of the transcription start site (TSS), where RNAPII normally transitions to its maximum processivity. We also identify an important role for INTS11, possibly in association with RNAPII CTD phospho-Tyr1, in repressing antisense transcription upstream of active promoters, as well as repressing transcription of genic regions near AsiSI-induced double-strand breaks.

Altogether, this study points toward a novel function of Integrator in promoting termination of incompetent RNAPII molecules while facilitating the transition to fully processive polymerase in order to enable efficient elongation.
]]></description>
<dc:creator>Rohban, S.</dc:creator>
<dc:creator>Rafiee, M.-R.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Luscombe, N.</dc:creator>
<dc:date>2023-02-17</dc:date>
<dc:identifier>doi:10.1101/2023.02.17.528960</dc:identifier>
<dc:title><![CDATA[Human Integrator provides a quality checkpoint during elongation to facilitate RNA polymerase II processivity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.17.528922v1?rss=1">
<title>
<![CDATA[
Discovery of multi-state gene cluster switches determining the adaptive mitochondrial and metabolic landscape of breast cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.17.528922v1?rss=1"
</link>
<description><![CDATA[
Adaptive metabolic switches are proposed to underlie conversions between cellular states during normal development as well as in cancer evolution, where they represent important therapeutic targets. However, the full spectrum, characteristics and regulation of existing metabolic switches are unknown. We propose that metabolic switches can be recognised by locating large alternating gene expression patterns and associate them with specific metabolic states. We developed a method to identify interspersed genesets by massive correlated biclustering (MCbiclust) and to predict their metabolic wiring. Testing the method on major breast cancer transcriptome datasets we discovered a series of gene sets with switch-like behaviour, predicting mitochondrial content, activity and central carbon fluxes in tumours associated with different switch positions. The predictions were experimentally validated by bioenergetic profiling and metabolic flux analysis of 13C-labelled substrates and were ultimately extended by geneset analysis to link metabolic alterations to cellular states, thus predicting tumour pathology, prognosis and chemosensitivity. The method is applicable to any large and heterogeneous transcriptome dataset to discover metabolic and associated pathophysiological states.

Statement of significanceWe present a novel method to identify the transcriptomic signatures of metabolic switches underlying divergent routes of cellular transformation. We illustrate the power of the method by stratifying breast cancer into metabolic subtypes, predicting their biology, architecture and clinical outcome.
]]></description>
<dc:creator>Menegollo, M.</dc:creator>
<dc:creator>Bentham, R. B.</dc:creator>
<dc:creator>Henriques, T.</dc:creator>
<dc:creator>Ng, S. Q.</dc:creator>
<dc:creator>Ren, Z.</dc:creator>
<dc:creator>Esculier, C.</dc:creator>
<dc:creator>Agarwal, S.</dc:creator>
<dc:creator>Tong, E.</dc:creator>
<dc:creator>Ilangovan, S.</dc:creator>
<dc:creator>Szabadkai, Z.</dc:creator>
<dc:creator>Suman, M.</dc:creator>
<dc:creator>Patani, N.</dc:creator>
<dc:creator>Ghanate, A.</dc:creator>
<dc:creator>Bryson, K.</dc:creator>
<dc:creator>Stein, R. C.</dc:creator>
<dc:creator>Yuneva, M.</dc:creator>
<dc:creator>Szabadkai, G.</dc:creator>
<dc:date>2023-02-18</dc:date>
<dc:identifier>doi:10.1101/2023.02.17.528922</dc:identifier>
<dc:title><![CDATA[Discovery of multi-state gene cluster switches determining the adaptive mitochondrial and metabolic landscape of breast cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.17.528925v1?rss=1">
<title>
<![CDATA[
Active mesh and neural network pipeline for cell aggregate segmentation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.17.528925v1?rss=1"
</link>
<description><![CDATA[
Segmenting cells within cellular aggregates in 3D is a growing challenge in cell biology, due to improvements in capacity and accuracy of microscopy techniques. Here we describe a pipeline to segment images of cell aggregates in 3D. The pipeline combines neural network segmentations with active meshes. We apply our segmentation method to cultured mouse mammary duct organoids imaged over 24 hours with oblique plane microscopy, a high-throughput light-sheet fluorescence microscopy technique. We show that our method can also be applied to images of mouse embryonic stem cells imaged with a spinning disc microscope. We segment individual cells based on nuclei and cell membrane fluorescent markers, and track cells over time. We describe metrics to quantify the quality of the automated segmentation. Our segmentation pipeline involves a Fiji plugin which implement active meshes deformation and allows a user to create training data, automatically obtain segmentation meshes from original image data or neural network prediction, and manually curate segmentation data to identify and correct mistakes. Our active meshes-based approach facilitates segmentation postprocessing, correction, and integration with neural network prediction.

Statement of significanceIn vitro culture of organ-like structures derived from stem cells, so-called organoids, allows to image tissue morphogenetic processes with high temporal and spatial resolution. Three-dimensional segmentation of cell shape in timelapse movies of these developing organoids is however a significant challenge. In this work, we propose an image analysis pipeline for cell aggregates that combines deep learning with active contour segmentations. This combination offers a flexible and efficient way to segment three-dimensional cell images, which we illustrate with by segmenting datasets of growing mammary gland organoids and mouse embryonic stem cells.
]]></description>
<dc:creator>Smith, M. B.</dc:creator>
<dc:creator>Salbreux, G.</dc:creator>
<dc:creator>Dunsby, C.</dc:creator>
<dc:creator>Sparks, H.</dc:creator>
<dc:creator>Almagro, J.</dc:creator>
<dc:creator>Behrens, A.</dc:creator>
<dc:creator>Chaigne, A.</dc:creator>
<dc:date>2023-02-21</dc:date>
<dc:identifier>doi:10.1101/2023.02.17.528925</dc:identifier>
<dc:title><![CDATA[Active mesh and neural network pipeline for cell aggregate segmentation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.20.529276v1?rss=1">
<title>
<![CDATA[
Pan-genotypic probe-based enrichment to improve efficiency of Hepatitis B virus sequencing 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.20.529276v1?rss=1"
</link>
<description><![CDATA[
Hepatitis B Virus (HBV) genome sequencing can be used to provide more complete genetic information at the population and individual level to shed light on the limitations of current interventions, and inform new strategies for elimination. HBV sequencing is challenging due to the partially dsDNA genome, high diversity, low viral loads and presence of large amounts of host genetic material in clinical samples. Here we describe the design and use of a pan-genotypic panel of 74 HBV specific capture-probes and nuclease treatment in improving sequencing efficiency. We processed 20 plasma samples (viral loads 1.98 to 4.07 log10, genotypes A-E) and three positive controls (human total brain RNA and bacteriophage lambda DNA) in triplicate to compare DNAse vs. RNAse vs. no nuclease treatment. We prepared libraries using the Takara Bio SMARTer Stranded Total RNA-Seq Kit v3, split the library in two, enriching half with the custom-designed probe panel and xGen Hybridization and Wash Kit (IDT), the other half was not enriched. Both libraries were sequenced on the NovaSeq6000 platform with 2x150nt paired-end reads. Capture resulted in a 47,970 fold increase in the number of reads mapped to the HBV genome in the "no nuclease" arm (243 HBV reads per million reads sequenced in the capture pool vs. 5x10-3 reads per million in the no-capture pool). Out of 20 samples, only 1 without capture generated HBV reads (viral load 3.89 log10 IU/ml) vs. 19 samples with capture. HBV sequence yield was increased in the capture arm and resulted in 2.30 log10 (95% confidence interval 1.99 - 2.48 log10) increase in HBV reads (per million reads sequenced) per log10 increase in viral load. The proportion of HBV reads increased a median of 12 fold with RNAse treatment. We developed a targeted pan-genotypic sequencing method using a custom panel of biotinylated oligos that increases the sequencing efficacy of HBV. This method will allow us to gain a better insight into HBV diversity.
]]></description>
<dc:creator>Lumley, S. F.</dc:creator>
<dc:creator>Jennings, D.</dc:creator>
<dc:creator>Waddilove, E.</dc:creator>
<dc:creator>Trebes, A.</dc:creator>
<dc:creator>Delphin, M.</dc:creator>
<dc:creator>Downs, L. O.</dc:creator>
<dc:creator>Macintyre-Crockett, G.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Chaudron, S.</dc:creator>
<dc:creator>de Lara, C.</dc:creator>
<dc:creator>Chai, H.</dc:creator>
<dc:creator>Maponga, T.</dc:creator>
<dc:creator>Martin, J.</dc:creator>
<dc:creator>Collier, J.</dc:creator>
<dc:creator>Ip, C.</dc:creator>
<dc:creator>Barnes, E.</dc:creator>
<dc:creator>Bonsall, D.</dc:creator>
<dc:creator>Piazza, P.</dc:creator>
<dc:creator>Ansari, M. A.</dc:creator>
<dc:creator>Matthews, P. C.</dc:creator>
<dc:date>2023-02-21</dc:date>
<dc:identifier>doi:10.1101/2023.02.20.529276</dc:identifier>
<dc:title><![CDATA[Pan-genotypic probe-based enrichment to improve efficiency of Hepatitis B virus sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.21.529250v1?rss=1">
<title>
<![CDATA[
Signalling-dependent refinement of cell fate choice during tissue remodelling 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.21.529250v1?rss=1"
</link>
<description><![CDATA[
How biological form emerges from cell fate decisions and tissue remodelling is a fundamental question in development biology. However, an understanding of how these processes operate side-by-side to set precise and robust patterns is largely missing. Here, we investigate this interplay during the process of vein refinement in the Drosophila pupal wing. By following reporters of signalling activity dynamically, together with tissue flows, we show that longitudinal vein refinement arises from a combination of local tissue deformation and cell fate adjustments controlled by a signalling network involving Notch, Dpp, and EGFR. Perturbing large-scale convergence and extension tissue flows does not affect vein refinement, showing that pre-patterned vein domains are able to intrinsically refine to the correct width. A minimal biophysical description taking into account key signalling interactions recapitulates the intrinsic tissue ability to establish a thin, regular vein independently of large-scale tissue flows. Supporting this prediction, artificial proveins optogenetically generated orthogonal to the axis of wing elongation refine against large-scale flows. Overall, we find that signalling-mediated updating of cell fate is a key contributor to reproducible patterning.
]]></description>
<dc:creator>Herszterg, S.</dc:creator>
<dc:creator>de Gennes, M.</dc:creator>
<dc:creator>Cicolini, S.</dc:creator>
<dc:creator>Huang, A.</dc:creator>
<dc:creator>Alexandre, C.</dc:creator>
<dc:creator>Smith, M. B.</dc:creator>
<dc:creator>Araujo, H.</dc:creator>
<dc:creator>Vincent, J.-P.</dc:creator>
<dc:creator>Salbreux, G.</dc:creator>
<dc:date>2023-02-22</dc:date>
<dc:identifier>doi:10.1101/2023.02.21.529250</dc:identifier>
<dc:title><![CDATA[Signalling-dependent refinement of cell fate choice during tissue remodelling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.24.529907v1?rss=1">
<title>
<![CDATA[
The level of Nck rather than N-WASP determines the rate of actin-based motility of Vaccinia 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.24.529907v1?rss=1"
</link>
<description><![CDATA[
Vaccinia virus exiting from host cells activates Src/Abl kinases to phosphorylate A36, an integral membrane viral protein. Phosphorylated A36 binds the adaptors Nck and Grb2 which recruit N-WASP to activate Arp2/3-driven actin polymerisation to promote viral spread. A36 also recruits intersectin, which enhances actin polymerization via AP-2/clathrin and Cdc42. How many viral and host molecules does vaccinia hijack to induce actin polymerization? To advance our quantitative understanding of this process, we now determined absolute numbers of the essential molecules in the vaccinia signalling network using fluorescent molecule counting approaches in live cells. There are 1156{+/-}120 A36 molecules on virus particles inducing actin polymerization in HeLa cells. This number, however, is over 2000 in mouse embryonic fibroblasts (MEFs), suggesting that A36 levels on the virion are not fixed. In MEFs, viruses recruit 1032{+/-}200 Nck and 434{+/-}10 N-WASP molecules, suggesting a ratio of 4:2:1 for the A36:Nck:N-WASP signalling network. Loss of A36 binding to either secondary adaptors Grb2 or intersectin results in a 1.3- and 2.5-fold reduction in Nck respectively. Curiously, despite recruiting comparable numbers of the Arp2/3 activator, N-WASP (245{+/-}26 and 276{+/-}66), these mutant viruses move at different speeds that inversely correlate with the number of Nck molecules. Our analysis has uncovered two unexpected new aspects of Vaccinia virus egress, numbers of the viral protein A36 can vary in the virion membrane and the velocity of virus movement depends on the levels of the adaptor protein Nck.
]]></description>
<dc:creator>Basant, A.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:date>2023-02-24</dc:date>
<dc:identifier>doi:10.1101/2023.02.24.529907</dc:identifier>
<dc:title><![CDATA[The level of Nck rather than N-WASP determines the rate of actin-based motility of Vaccinia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.27.530267v1?rss=1">
<title>
<![CDATA[
Notochord and axial progenitor generation by timely BMP and NODAL inhibition during vertebrate trunk formation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.27.530267v1?rss=1"
</link>
<description><![CDATA[
The formation of the vertebrate body involves the coordinated and progressive production of trunk tissues from progenitors located in the posterior of the embryo. In vitro models based on pluripotent stem cells (PSCs) replicate aspects of this process, but they lack some tissue components normally present in the trunk. Most strikingly, the notochord, a hallmark of chordates and the source of midline signals that pattern surrounding tissues, is absent from current models of human trunk formation. To investigate how trunk tissue is formed, we performed single-cell transcriptomic analysis of chick embryos. This delineated molecularly discrete progenitor populations, which we spatially locate in the embryo, compare across species, and relate to signalling activity. Guided by this map, we determined how differentiating human PSCs develop a stereotypical spatial organization of tissue types. We found that LATS1/2 repression of YAP activity, in conjunction with FGF-mediated MAPK activation, induced the transcription factor Bra/TBXT and facilitated WNT signaling. In addition, inhibiting a WNT-induced NODAL and BMP signaling cascade at the appropriate time regulated the proportions of different tissue types produced, including notochordal cells. We used this information to create an integrated 3D model of human gastrulation undergoing morphogenetic movements to produce elongated structures with a notochord and spatially patterned neural tissue formation. Together the data provide insight into the mechanisms responsible for the formation of the tissues that comprise the vertebrate trunk and pave the way for future studies of patterning in a tissue-like environment.
]]></description>
<dc:creator>Rito, T.</dc:creator>
<dc:creator>Libby, A. R.</dc:creator>
<dc:creator>Demuth, M.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:date>2023-02-28</dc:date>
<dc:identifier>doi:10.1101/2023.02.27.530267</dc:identifier>
<dc:title><![CDATA[Notochord and axial progenitor generation by timely BMP and NODAL inhibition during vertebrate trunk formation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.03.06.531353v1?rss=1">
<title>
<![CDATA[
Peroxisomal compartmentalization of amino acid biosynthesis reactions imposes an upper limit on compartment size 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.03.06.531353v1?rss=1"
</link>
<description><![CDATA[
Cellular metabolism relies on just a few redox cofactors. Selective compartmentalization may prevent competition between metabolic reactions requiring the same cofactor. Is such compartmentalization necessary for optimal cell function? Is there an optimal compartment size? Here we probe these fundamental questions using peroxisomal compartmentalization of the last steps of lysine and histidine biosynthesis in the fission yeast Schizosaccharomyces japonicus. We show that compartmentalization of these NAD+ dependent reactions together with a dedicated NADH/NAD+ recycling enzyme supports optimal growth when an increased demand for anabolic reactions taxes cellular redox balance. In turn, compartmentalization constrains the size of individual organelles, with larger peroxisomes accumulating all the required enzymes but unable to support both biosynthetic reactions at the same time. We propose that compartmentalized biosynthetic reactions are sensitive to the size of the compartment, likely due to scaling-dependent changes within the system, such as enzyme packing density.
]]></description>
<dc:creator>Gu, Y.</dc:creator>
<dc:creator>Alam, S.</dc:creator>
<dc:creator>Oliferenko, S.</dc:creator>
<dc:date>2023-03-07</dc:date>
<dc:identifier>doi:10.1101/2023.03.06.531353</dc:identifier>
<dc:title><![CDATA[Peroxisomal compartmentalization of amino acid biosynthesis reactions imposes an upper limit on compartment size]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.03.07.531515v1?rss=1">
<title>
<![CDATA[
ORC1 binds to cis-transcribed RNAs for efficient activation of replication origins 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.03.07.531515v1?rss=1"
</link>
<description><![CDATA[
Cells must coordinate the activation of thousands of replication origins dispersed throughout their genome. Active transcription is known to favor the formation of mammalian origins, although the role that RNA plays in this process remains unclear. We show that the ORC1 subunit of the human Origin Recognition Complex interacts with RNAs transcribed from genes with origins in their transcription start sites (TSSs), displaying a positive correlation between RNA binding and origin activity. RNA depletion, or the use of ORC1 RNA-binding mutant, result in inefficient activation of proximal origins, linked to impaired ORC1 chromatin release. ORC1 RNA binding activity resides in its intrinsically disordered region, involved in intra- and inter-molecular interactions, regulation by phosphorylation, and phase-separation. We show that RNA binding favors ORC1 chromatin release, by regulating its phosphorylation and subsequent degradation. We propose that fluctuating concentrations of RNA during the cell cycle may play a sequential role in controlling origins through interaction with this flexible region of ORC1. Our results unveil a novel non-coding function of RNA as a dynamic component of the chromatin, orchestrating the activation of replication origins.

One sentence summaryThe human origin recognition complex subunit 1 ORC1, binds to RNAs transcribed from genes with origins of replication at the TSS, which is required for optimal origin activation.
]]></description>
<dc:creator>Mas, A. M.</dc:creator>
<dc:creator>Goni, E.</dc:creator>
<dc:creator>Ruiz de los Mozos, I.</dc:creator>
<dc:creator>Arcas, A.</dc:creator>
<dc:creator>Statello, L.</dc:creator>
<dc:creator>Gonzalez, J.</dc:creator>
<dc:creator>Blazquez, L.</dc:creator>
<dc:creator>Lee, W. T. C.</dc:creator>
<dc:creator>Gupta, D.</dc:creator>
<dc:creator>Sejas, A.</dc:creator>
<dc:creator>Hoshina, S.</dc:creator>
<dc:creator>Armaos, A.</dc:creator>
<dc:creator>Tartaglia, G. G.</dc:creator>
<dc:creator>Waga, S.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Rothenberg, E.</dc:creator>
<dc:creator>Gomez, M.</dc:creator>
<dc:creator>Huarte, M.</dc:creator>
<dc:date>2023-03-07</dc:date>
<dc:identifier>doi:10.1101/2023.03.07.531515</dc:identifier>
<dc:title><![CDATA[ORC1 binds to cis-transcribed RNAs for efficient activation of replication origins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.03.07.531437v1?rss=1">
<title>
<![CDATA[
Dissecting the subcellular forces sculpting early C. elegans embryos 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.03.07.531437v1?rss=1"
</link>
<description><![CDATA[
Embryo shape is determined by individual cell mechanics, intercellular interaction strength, and geometrical constraints. Models based on surface tensions at cell interfaces can predict 3D static cellular arrangements within aggregates. However, predicting the dynamics of such arrangements is challenging due to difficulties in measuring temporal changes in tensions. Here, we characterise the spatiotemporal changes in cellular tensions shaping the early nematode embryo using AFM, live microscopy, and tension inference. Using excoriated embryos, we validate a hybrid inference pipeline that calibrates relative inferred tensions temporally using cortical myosin enrichment and absolute tensions using AFM measurements. Applied to embryos within their native shell, we infer a spatiotemporal map of absolute tensions, revealing that ABa, ABp, and EMS compaction is driven by increased tension at free surfaces, while P2s initial exclusion is due to high tension at intercellular contacts. We uncover a direct and non-affine contribution of cadherins to cell-cell contact tension, comparable to cadherins indirect contribution via actomyosin regulation.

HighlightsO_LIP lineage cells have lower cortical tensions than AB lineage cells
C_LIO_LIEnrichment of Myosin-II at the cell cortex is a good predictor of cell-medium tension but is not sufficient to determine tension at cell-cell contacts.
C_LIO_LIMyosin-informed tension inference allows determination of the spatiotemporal evolution of all surface tensions within the embryo.
C_LIO_LIABa, ABp, and EMS compact due to high tensions at their cell-medium interfaces compared to their cell-cell interfaces, while P2 is initially excluded due to high cell-cell contact tensions.
C_LIO_LICadherins contribute directly in a non-linear way by reducing cell-cell contact tension by nearly 50%.
C_LI

Open AccessFor the purpose of Open Access, the author has applied a CC BY public copyright license to any Author Accepted Manuscript version arising from this submission.
]]></description>
<dc:creator>Yamamoto, K.</dc:creator>
<dc:creator>Ichbiah, S.</dc:creator>
<dc:creator>Pinto, J.</dc:creator>
<dc:creator>Delbary, F.</dc:creator>
<dc:creator>Goehring, N.</dc:creator>
<dc:creator>Turlier, H.</dc:creator>
<dc:creator>Charras, G.</dc:creator>
<dc:date>2023-03-09</dc:date>
<dc:identifier>doi:10.1101/2023.03.07.531437</dc:identifier>
<dc:title><![CDATA[Dissecting the subcellular forces sculpting early C. elegans embryos]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.03.09.531882v1?rss=1">
<title>
<![CDATA[
Nondestructive Spatial Lipidomics for Glioma Classification 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.03.09.531882v1?rss=1"
</link>
<description><![CDATA[
Mapping the molecular composition of tissues using spatial biology provides high-content information for molecular diagnostics. However, spatial biology approaches require invasive procedures to collect samples and destroy the investigated tissue, limiting the extent of analysis, particularly for highly functional tissues such as those of the brain. To address these limitations, we developed a workflow to harvest biomolecules from brain tissues using nanoneedles and characterise the distribution of lipids using desorption electrospray ionization mass spectrometry imaging. The nanoneedles preserved the original tissue while harvesting a reliable molecular profile and retaining the original lipid distribution for mouse and human brain samples, accurately outlining the morphology of key regions within the brain and tumour lesions. The deep neural network analysis of a cohort containing 23 human glioma biopsies showed that nanoneedle samples maintain the molecular signatures required to accurately classify disease state. Thus, nanoneedles provide a route for tissue-preserving spatial lipidomic and molecular diagnostics.
]]></description>
<dc:creator>Martella, D. A.</dc:creator>
<dc:creator>Rose, L. A.</dc:creator>
<dc:creator>Rouatbi, N.</dc:creator>
<dc:creator>Gu, C.</dc:creator>
<dc:creator>Caprettini, V.</dc:creator>
<dc:creator>Jensen, M.</dc:creator>
<dc:creator>Hagemann, C.</dc:creator>
<dc:creator>Serio, A.</dc:creator>
<dc:creator>Al-Jamal, K.</dc:creator>
<dc:creator>Parsons, M.</dc:creator>
<dc:creator>Bergholt, M.</dc:creator>
<dc:creator>Brennan, P.</dc:creator>
<dc:creator>Zaritski, A.</dc:creator>
<dc:creator>Chiappini, C.</dc:creator>
<dc:date>2023-03-12</dc:date>
<dc:identifier>doi:10.1101/2023.03.09.531882</dc:identifier>
<dc:title><![CDATA[Nondestructive Spatial Lipidomics for Glioma Classification]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.03.09.531725v1?rss=1">
<title>
<![CDATA[
RNA allelic frequencies of somatic mutations encode substantial functional information in cancers 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.03.09.531725v1?rss=1"
</link>
<description><![CDATA[
A central goal of cancer research is the identification of cancer genes that drive tumour growth and progression. Existing approaches to this problem typically leverage frequentist approaches based on patterns of somatic mutagenesis in DNA. Here, we interrogate RNA variant allele frequencies to identify putative cancer genes with a novel computational tool, RVdriver, from bulk genomic-transcriptomic data within 7,948 paired exomes and transcriptomes across 30 cancer types. An elevated RNA VAF reflects a signal from multiple biological features: clonal mutations; mutations retained or gained during somatic copy-number alterations; mutations favoured by allele-specific expression; and mutations in genes expressed preferentially by the tumour compartment of admixed bulk samples. RVdriver, a statistical approach that classifies RNA VAFs of nonsynonymous mutations relative to a synonymous mutation background, leverages this information to identify known, as well as putatively novel, cancer genes, with comparable performance to DNA-based approaches. Furthermore, we demonstrate RNA VAFs of individual mutations are able to distinguish  driver from  passenger mutations within established cancer genes. Low-RNA VAF EGFR mutations otherwise annotated as drivers of glioblastoma by DNA tools harbour a phenotype of reduced EGFR signalling, whilst high-RNA VAF KDM6A mutations otherwise annotated as passengers exhibit a driver-like H3K27me3 expression profile, demonstrating the value of our approach in phenotyping tumours. Overall, our study showcases a novel approach for cancer gene discovery, and highlights the potential value of multi-omic and systems-biology approaches in finding novel therapeutic vulnerabilities in cancer to bring about patient benefit.
]]></description>
<dc:creator>Black, J. R. M.</dc:creator>
<dc:creator>Jones, T. P.</dc:creator>
<dc:creator>Martinez-Ruiz, C.</dc:creator>
<dc:creator>Litovchenko, M.</dc:creator>
<dc:creator>Puttick, C.</dc:creator>
<dc:creator>McGranahan, N.</dc:creator>
<dc:date>2023-03-11</dc:date>
<dc:identifier>doi:10.1101/2023.03.09.531725</dc:identifier>
<dc:title><![CDATA[RNA allelic frequencies of somatic mutations encode substantial functional information in cancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.03.12.531945v1?rss=1">
<title>
<![CDATA[
Metabolic profiling stratifies colorectal cancer and reveals adenosylhomocysteinase as a therapeutic target 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.03.12.531945v1?rss=1"
</link>
<description><![CDATA[
With colorectal cancer (CRC) being the second most common cause of cancer-related deaths worldwide1, there is an urgent need for better diagnostic tools and new, more targeted therapies. Here we used genetically engineered mouse models (GEMMs), and multimodal mass spectrometry-based metabolomics to study the impact of common genetic drivers of CRC on the metabolic landscape of the intestine. We show that unsupervised metabolic profiling can stratify intestinal tissues according to underlying genetic alterations, and use mass spectrometry imaging (MSI) to identify tumour, stromal and normal adjacent tissues. By identifying ions that drive variation between normal and transformed tissues, we found dysregulation of the methionine cycle to be a hallmark of APC-mutant CRC, and propose one of its enzymes, i.e. adenosylhomocysteinase (AHCY), as a new therapeutic target. Collectively, we show that the profound genotype-dependent alterations in both lipid and small molecule metabolism in CRC may be exploited for tissue classification with no need for ion identification, and we applied further data analysis to expose a novel metabolic vulnerability of CRC.
]]></description>
<dc:creator>Vande Voorde, J.</dc:creator>
<dc:creator>Najumudeen, A. K.</dc:creator>
<dc:creator>Steven, R. T.</dc:creator>
<dc:creator>Nikula, C. J.</dc:creator>
<dc:creator>Dexter, A.</dc:creator>
<dc:creator>Zeiger, L. B.</dc:creator>
<dc:creator>Elia, E. A.</dc:creator>
<dc:creator>Nassif, A.</dc:creator>
<dc:creator>Gonzalez-Fernandez, A.</dc:creator>
<dc:creator>Murta, T.</dc:creator>
<dc:creator>Gillespie, M.</dc:creator>
<dc:creator>Ford, C. A.</dc:creator>
<dc:creator>Lannagan, T. R. M.</dc:creator>
<dc:creator>Vlahov, N.</dc:creator>
<dc:creator>Ridgway, R. A.</dc:creator>
<dc:creator>Nixon, C.</dc:creator>
<dc:creator>Gilroy, K.</dc:creator>
<dc:creator>Gay, D. M.</dc:creator>
<dc:creator>Burton, A.</dc:creator>
<dc:creator>Yan, B.</dc:creator>
<dc:creator>Sellers, K.</dc:creator>
<dc:creator>Wu, V.</dc:creator>
<dc:creator>Xiang, Y.</dc:creator>
<dc:creator>Shokry, E.</dc:creator>
<dc:creator>Clark, W.</dc:creator>
<dc:creator>Li, V. S. W.</dc:creator>
<dc:creator>Barry, S. T.</dc:creator>
<dc:creator>Goodwin, R. J. A.</dc:creator>
<dc:creator>Takats, Z.</dc:creator>
<dc:creator>Maddocks, O. D. K.</dc:creator>
<dc:creator>Sumpton, D.</dc:creator>
<dc:creator>Yuneva, M. O.</dc:creator>
<dc:creator>Campbell, A. D.</dc:creator>
<dc:creator>Bunch, J.</dc:creator>
<dc:creator>Sansom, O. J.</dc:creator>
<dc:date>2023-03-12</dc:date>
<dc:identifier>doi:10.1101/2023.03.12.531945</dc:identifier>
<dc:title><![CDATA[Metabolic profiling stratifies colorectal cancer and reveals adenosylhomocysteinase as a therapeutic target]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.03.13.532312v1?rss=1">
<title>
<![CDATA[
A malaria parasite phospholipase facilitates efficient asexual blood stage egress 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.03.13.532312v1?rss=1"
</link>
<description><![CDATA[
Malaria parasite release (egress) from host red blood cells involves parasite-mediated membrane poration and rupture, thought to involve membrane-lytic effector molecules such as perforin-like proteins and/or phospholipases. With the aim of identifying these effectors, we disrupted the expression of two Plasmodium falciparum perforin-like proteins simultaneously and showed that they have no essential roles during blood stage egress. Proteomic profiling of parasite proteins discharged into the parasitophorous vacuole (PV) just prior to egress detected the presence in the PV of a lecithin:cholesterol acyltransferase (LCAT; PF3D7_0629300). Conditional ablation of LCAT resulted in abnormal egress and a reduced replication rate. Lipidomic profiles showed drastic changes in several phosphatidylserine and acylphosphatidylglycerol species during egress. We thus show that, in addition to its previously demonstrated role in liver stage merozoite egress, LCAT is required to facilitate efficient egress in asexual blood stage malaria parasites.

Author SummaryMalaria kills over half a million people every year worldwide. It is caused by a single-celled parasite called Plasmodium falciparum that grows and multiplies within a bounding vacuole, inside red blood cells of the infected individuals. Following each round of multiplication, the infected cell is ruptured in a process known as egress to release a new generation of parasites. Egress is required for the disease to progress and is orchestrated by the parasite. The parasite sends out various molecules to puncture and destroy the membranes of the vacuole and the red blood cell. However, little is known about these molecules. In this work, we set out to identify these molecules by using genetic and proteomics approaches. We screened the molecules the parasite sends out during egress and identified a parasite enzyme called LCAT present in the vacuole. Our experiments found that mutant parasites that were unable to make LCAT clumped together and could not escape the infected cell properly. As a result, we saw a reduction in the rate at which these parasites spread through the red blood cells. Taken together, our findings suggest that P. falciparum needs LCAT to efficiently break out of red blood cells.
]]></description>
<dc:creator>Ramaprasad, A.</dc:creator>
<dc:creator>Burda, P.-C.</dc:creator>
<dc:creator>Koussis, K.</dc:creator>
<dc:creator>Thomas, J. A.</dc:creator>
<dc:creator>Pietsch, E.</dc:creator>
<dc:creator>Calvani, E.</dc:creator>
<dc:creator>Howell, S. A.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Snijders, A. P.</dc:creator>
<dc:creator>Gilberger, T. W.</dc:creator>
<dc:creator>Blackman, M. J.</dc:creator>
<dc:date>2023-03-13</dc:date>
<dc:identifier>doi:10.1101/2023.03.13.532312</dc:identifier>
<dc:title><![CDATA[A malaria parasite phospholipase facilitates efficient asexual blood stage egress]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.03.14.532615v1?rss=1">
<title>
<![CDATA[
Insufficient evidence for natural selection associated with the Black Death 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.03.14.532615v1?rss=1"
</link>
<description><![CDATA[
Klunk et al. analyzed ancient DNA data from individuals in London and Denmark before, during and after the Black Death [1], and argued that allele frequency changes at immune genes were too large to be produced by random genetic drift and thus must reflect natural selection. They also identified four specific variants that they claimed show evidence of selection including at ERAP2, for which they estimate a selection coefficient of 0.39-several times larger than any selection coefficient on a common human variant reported to date. Here we show that these claims are unsupported for four reasons. First, the signal of enrichment of large allele frequency changes in immune genes comparing people in London before and after the Black Death disappears after an appropriate randomization test is carried out: the P value increases by ten orders of magnitude and is no longer significant. Second, a technical error in the estimation of allele frequencies means that none of the four originally reported loci actually pass the filtering thresholds. Third, the filtering thresholds do not adequately correct for multiple testing. Finally, in the case of the ERAP2 variant rs2549794, which Klunk et al. show experimentally may be associated with a host interaction with Y. pestis, we find no evidence of significant frequency change either in the data that Klunk et al. report, or in published data spanning 2,000 years. While it remains plausible that immune genes were subject to natural selection during the Black Death, the magnitude of this selection and which specific genes may have been affected remains unknown.
]]></description>
<dc:creator>Barton, A. R.</dc:creator>
<dc:creator>Santander, C. G.</dc:creator>
<dc:creator>Skoglund, P.</dc:creator>
<dc:creator>Moltke, I.</dc:creator>
<dc:creator>Reich, D.</dc:creator>
<dc:creator>Mathieson, I.</dc:creator>
<dc:date>2023-03-15</dc:date>
<dc:identifier>doi:10.1101/2023.03.14.532615</dc:identifier>
<dc:title><![CDATA[Insufficient evidence for natural selection associated with the Black Death]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.03.30.534739v1?rss=1">
<title>
<![CDATA[
Dietary environmental factors shape the defence against infection with Cryptosporidium 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.03.30.534739v1?rss=1"
</link>
<description><![CDATA[
Cryptosporidium is a leading cause of diarrheal-related deaths in children, especially in resource-poor settings. It also targets the immunocompromised, chronically infecting people living with HIV and primary immunodeficiencies. There is no vaccine or effective treatment. While it is known from human cases and animal models that CD4+ T-cells play a role in curbing Cryptosporidium, the role of CD8+ cells remains to be defined. Using a Cryptosporidium tyzzeri mouse model, we show that gut-resident CD8+ intraepithelial lymphocytes (IELs) confer resistance to parasite growth. CD8+ IELs express, and are dependent on, the ligand-dependent transcription factor aryl hydrocarbon receptor (AHR). AHR deficiency reduced CD8+ IELs, decreased their cytotoxicity, and worsened infection. Transfer of CD8+ IELs rescued severely immunodeficient mice from death following Cryptosporidium challenge. Finally, dietary supplementation of the AHR pro-ligand indole-3-carbinol to new-born mice promoted resistance to infection. Therefore, common dietary metabolites augment the host immune response to cryptosporidiosis, protecting against disease.
]]></description>
<dc:creator>Sateriale, A.</dc:creator>
<dc:creator>Maradana, M. R.</dc:creator>
<dc:creator>Marzook, N. B.</dc:creator>
<dc:creator>Diaz Perez, O.</dc:creator>
<dc:creator>Mkandawire, T.</dc:creator>
<dc:creator>Diny, N. L.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Liebert, A.</dc:creator>
<dc:creator>Shah, K.</dc:creator>
<dc:creator>Tolaini, M.</dc:creator>
<dc:creator>Kvac, M.</dc:creator>
<dc:creator>Stockinger, B.</dc:creator>
<dc:date>2023-04-01</dc:date>
<dc:identifier>doi:10.1101/2023.03.30.534739</dc:identifier>
<dc:title><![CDATA[Dietary environmental factors shape the defence against infection with Cryptosporidium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.04.06.535808v1?rss=1">
<title>
<![CDATA[
The minus end depolymerase KIF2A drives flux-like treadmilling of γTuRC-uncapped microtubules 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.04.06.535808v1?rss=1"
</link>
<description><![CDATA[
During mitosis, a functional spindle requires high microtubule turnover. Such turnover is highlighted by the multiple functions of spindle poles, where microtubule minus ends are concentrated, and where microtubule nucleation and depolymerization happen side by side. How these seemingly antagonistic processes are coordinated during poleward microtubule flux is not understood. Here we reconstitute this coordination in vitro combining different pole localized activities. We find that the spindle pole-localized kinesin-13 KIF2A is a microtubule minus-end depolymerase, in contrast to its paralog MCAK. Due to its asymmetric activity, KIF2A still allows microtubule nucleation by plus-end growth from the {gamma}-tubulin ring complex ({gamma}TuRC), which in turn serves as a protective cap that shields the minus end against KIF2A binding. Efficient {gamma}TuRC-uncapping requires the combined action of KIF2A and a microtubule severing enzyme, then leading to treadmilling of the uncapped microtubule driven by KIF2A. Together these results provide insight into the molecular mechanisms by which a minimal protein module coordinates microtubule nucleation and depolymerization at spindle poles consistent with their role in poleward microtubule flux.
]]></description>
<dc:creator>Henkin, G.</dc:creator>
<dc:creator>Brito, C.</dc:creator>
<dc:creator>Thomas, C.</dc:creator>
<dc:creator>Surrey, T.</dc:creator>
<dc:date>2023-04-06</dc:date>
<dc:identifier>doi:10.1101/2023.04.06.535808</dc:identifier>
<dc:title><![CDATA[The minus end depolymerase KIF2A drives flux-like treadmilling of γTuRC-uncapped microtubules]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.04.10.536195v1?rss=1">
<title>
<![CDATA[
Spatiotemporal Genomic Profiling of Intestinal Metaplasia Reveals Clonal Dynamics of Gastric Cancer Progression 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.04.10.536195v1?rss=1"
</link>
<description><![CDATA[
Intestinal metaplasia (IM) is a pre-malignant condition of the gastric mucosa associated with increased gastric cancer (GC) risk. We analyzed 1256 gastric samples (1152 IMs) from 692 subjects through a prospective 10-year study. We identified 26 IM driver genes in diverse pathways including chromatin regulation (ARID1A) and intestinal homeostasis (SOX9), largely occurring as small clonal events. Analysis of clonal dynamics between and within subjects, and also longitudinally across time, revealed that IM clones are likely transient but increase in size upon progression to dysplasia, with eventual transmission of somatic events to paired GCs. Single-cell and spatial profiling highlighted changes in tissue ecology and lineage heterogeneity in IM, including an intestinal stem-cell dominant cellular compartment linked to early malignancy. Expanded transcriptome profiling revealed expression-based molecular subtypes of IM, including a body-resident "pseudoantralized" subtype associated with incomplete histology, antral/intestinal cell types, ARID1A mutations, inflammation, and microbial communities normally associated with the healthy oral tract. We demonstrate that combined clinical- genomic models outperform clinical-only models in predicting IMs likely to progress. Our results raise opportunities for GC precision prevention and interception by highlighting strategies for accurately identifying IM patients at high GC risk and a role for microbial dysbiosis in IM progression.
]]></description>
<dc:creator>Huang, K. K.</dc:creator>
<dc:creator>Ma, H.</dc:creator>
<dc:creator>Uchihara, T.</dc:creator>
<dc:creator>Sheng, T.</dc:creator>
<dc:creator>Chong, R. H. H.</dc:creator>
<dc:creator>Zhu, F.</dc:creator>
<dc:creator>Srivastava, S.</dc:creator>
<dc:creator>Tay, S. T.</dc:creator>
<dc:creator>Sundar, R.</dc:creator>
<dc:creator>Tan, A. L. K.</dc:creator>
<dc:creator>Ong, X.</dc:creator>
<dc:creator>Lee, M.</dc:creator>
<dc:creator>Ho, S. W. T.</dc:creator>
<dc:creator>Lesluyes, T.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>Chua, J. S.</dc:creator>
<dc:creator>Ramnarayanan, K.</dc:creator>
<dc:creator>Ang, T. L.</dc:creator>
<dc:creator>Khor, C.</dc:creator>
<dc:creator>Lee, J. W. J.</dc:creator>
<dc:creator>Tsao, S. K. K.</dc:creator>
<dc:creator>Teh, M.</dc:creator>
<dc:creator>Chung, H.</dc:creator>
<dc:creator>So, J. B. Y.</dc:creator>
<dc:creator>Yeoh, K. G.</dc:creator>
<dc:creator>Tan, P.</dc:creator>
<dc:date>2023-04-11</dc:date>
<dc:identifier>doi:10.1101/2023.04.10.536195</dc:identifier>
<dc:title><![CDATA[Spatiotemporal Genomic Profiling of Intestinal Metaplasia Reveals Clonal Dynamics of Gastric Cancer Progression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.04.14.536403v1?rss=1">
<title>
<![CDATA[
FOXL2 interaction with different binding partners regulates the dynamics of granulosa cell differentiation across ovarian development 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.04.14.536403v1?rss=1"
</link>
<description><![CDATA[
The transcription factor FOXL2 is required in ovarian somatic cells for female fertility. Differential timing of Foxl2 deletion, in embryonic versus adult mouse ovary, leads to distinctive outcomes suggesting different roles across development. Here, we comprehensively investigated FOXL2s role through a multi-omics approach to characterise gene expression dynamics and chromatin accessibility changes, coupled with genome-wide identification of FOXL2 targets and on-chromatin interacting partners in granulosa cells across ovarian development. We found that FOXL2 regulates more targets postnatally, through interaction with factors regulating primordial follicle activation (PFA) and steroidogenesis. Deletion of one interactor, Ubiquitin specific protease 7 (USP7), induces PFA blockage, impaired ovary development and sterility. Our datasets constitute a comprehensive resource for exploration of the molecular mechanisms of ovarian development and causes of female infertility.
]]></description>
<dc:creator>Migale, R.</dc:creator>
<dc:creator>Neumann, M.</dc:creator>
<dc:creator>Mitter, R.</dc:creator>
<dc:creator>Rafiee, M.-R.</dc:creator>
<dc:creator>Wood, S.</dc:creator>
<dc:creator>Olsen, J.</dc:creator>
<dc:creator>Lovell-Badge, R.</dc:creator>
<dc:date>2023-04-14</dc:date>
<dc:identifier>doi:10.1101/2023.04.14.536403</dc:identifier>
<dc:title><![CDATA[FOXL2 interaction with different binding partners regulates the dynamics of granulosa cell differentiation across ovarian development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.04.13.536740v1?rss=1">
<title>
<![CDATA[
In vivo CRISPR screen identifies KRAS-induced COX-2 as a driver of immune evasion and immunotherapy resistance in lung cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.04.13.536740v1?rss=1"
</link>
<description><![CDATA[
Oncogenic KRAS impairs anti-tumour immune responses, but effective strategies to combine KRAS inhibitors and immunotherapies have so far proven elusive. In vivo CRISPR-Cas9 screening in an immunogenic murine lung cancer model identifies mechanisms by which oncogenic KRAS promotes immune evasion, most notably expression of immunosuppressive cyclooxygenase-2 (COX-2) in cancer cells. Oncogenic KRAS was a potent inducer of COX-2 in both mouse and human lung cancer which was suppressed using KRAS inhibitors. COX-2 acting via prostaglandin E2 (PGE2) promotes resistance to immune checkpoint blockade (ICB) in both mouse and human lung adenocarcinoma. Targeting COX-2/PGE2 remodelled the tumour microenvironment by inducing pro-inflammatory polarisation of myeloid cells and influx of activated cytotoxic CD8+ T cells, which increased the efficacy of ICB. Restoration of COX-2 expression contributed to tumour relapse after prolonged KRAS inhibition. We propose testing COX-2/PGE2 pathway inhibitors in combination with KRAS G12C inhibition or ICB in patients with KRAS-mutant lung cancer.
]]></description>
<dc:creator>Boumelha, J.</dc:creator>
<dc:creator>de Castro, A.</dc:creator>
<dc:creator>Bah, N.</dc:creator>
<dc:creator>Cha, H.</dc:creator>
<dc:creator>de Carne Trecesson, S.</dc:creator>
<dc:creator>Rana, S.</dc:creator>
<dc:creator>Anastasiou, P.</dc:creator>
<dc:creator>Mugarza, E.</dc:creator>
<dc:creator>Moore, C.</dc:creator>
<dc:creator>Goldstone, R.</dc:creator>
<dc:creator>East, P.</dc:creator>
<dc:creator>Litchfield, K.</dc:creator>
<dc:creator>Lee, S.-H.</dc:creator>
<dc:creator>Molina-Arcas, M.</dc:creator>
<dc:creator>Downward, J.</dc:creator>
<dc:date>2023-04-14</dc:date>
<dc:identifier>doi:10.1101/2023.04.13.536740</dc:identifier>
<dc:title><![CDATA[In vivo CRISPR screen identifies KRAS-induced COX-2 as a driver of immune evasion and immunotherapy resistance in lung cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.04.18.537386v1?rss=1">
<title>
<![CDATA[
A palmitoyl transferase chemical genetic system to map ZDHHC-specific S-acylation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.04.18.537386v1?rss=1"
</link>
<description><![CDATA[
The 23 human ZDHHC S-acyltransferases catalyze long-chain S-acylation at cysteine residues across an extensive network of hundreds of proteins important for normal physiology or dysregulated in disease. Here we present a technology platform to directly map the protein substrates of a specific ZDHHC for the first time at the whole proteome level, in intact cells. Structure-guided engineering of paired ZDHHC  hole mutants and  bumped chemically tagged fatty acid probes enabled probe transfer to specific protein substrates with excellent selectivity over wild type ZDHHCs. Chemical genetic systems were exemplified for five ZDHHCs (3, 7, 11, 15 and 20), and applied to generate the first de novo ZDHHC substrate profiles, identifying >300 unique and shared substrates across multiple cell lines and S-acylation sites for novel functionally diverse substrates. We expect that this powerful and versatile platform will open a new window on S-acylation biology for a wide range of models and organisms.
]]></description>
<dc:creator>Ocasio, C. A.</dc:creator>
<dc:creator>Baggelaar, M. P.</dc:creator>
<dc:creator>Sipthorp, J.</dc:creator>
<dc:creator>Losada de la Lastra, A.</dc:creator>
<dc:creator>Volaric, J.</dc:creator>
<dc:creator>Soudy, C.</dc:creator>
<dc:creator>Storck, E. M.</dc:creator>
<dc:creator>Palma-Duran, S. A.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Tomic, G.</dc:creator>
<dc:creator>Carr, L.</dc:creator>
<dc:creator>Downward, J.</dc:creator>
<dc:creator>Eggert, U.</dc:creator>
<dc:creator>Tate, E. W.</dc:creator>
<dc:date>2023-04-18</dc:date>
<dc:identifier>doi:10.1101/2023.04.18.537386</dc:identifier>
<dc:title><![CDATA[A palmitoyl transferase chemical genetic system to map ZDHHC-specific S-acylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.04.17.537276v1?rss=1">
<title>
<![CDATA[
ER-dependent membrane repair of mycobacteria-induced vacuole damage 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.04.17.537276v1?rss=1"
</link>
<description><![CDATA[
Several intracellular pathogens, such as Mycobacterium tuberculosis, damage endomembranes to access the cytosol and subvert innate immune responses. The host counteracts endomembrane damage by recruiting repair machineries that retain the pathogen inside the vacuole.

Here, we show that the endoplasmic reticulum (ER)-Golgi protein oxysterol binding protein (OSBP) and its Dictyostelium discoideum homologue OSBP8 are recruited to the Mycobacterium-containing vacuole (MCV) after ESX-1-dependent membrane damage. Lack of OSBP8 causes a hyperaccumulation of phosphatidylinositol-4-phosphate (PI4P) on the MCV and decreased cell viability. OSBP8-depleted cells had reduced lysosomal and degradative capabilities of their vacuoles that favoured mycobacterial growth. In agreement with a function of OSBP8 in membrane repair, human macrophages infected with M. tuberculosis recruited OSBP in an ESX-1 dependent manner. These findings identified an ER-dependent repair mechanism for restoring MCVs in which OSBP8 functions to equilibrate PI4P levels on damaged membranes.

ImportanceTuberculosis still remains a global burden and is one of the top infectious diseases from a single pathogen. Mycobacterium tuberculosis, the causative agent, has perfected many ways to replicate and persist within its host. While mycobacteria induce vacuole damage to evade the toxic environment and eventually escape into the cytosol, the host recruits repair machineries to restore the MCV membrane. However, how lipids are delivered for membrane repair is poorly understood. Using advanced fluorescence imaging and volumetric correlative approaches, we demonstrate that this involves the recruitment of the ER-Golgi lipid transfer protein OSBP8 in the D. discoideum/ M. marinum system. Strikingly, depletion of OSBP8 affects lysosomal function accelerating mycobacterial growth. This indicates that an ER-dependent repair pathway constitutes a host defence mechanism against intracellular pathogens such as M. tuberculosis.
]]></description>
<dc:creator>Anand, A.</dc:creator>
<dc:creator>Mazur, A.-C.</dc:creator>
<dc:creator>Rosell-Arevalo, P.</dc:creator>
<dc:creator>Franzkoch, R.</dc:creator>
<dc:creator>Breitsprecher, L.</dc:creator>
<dc:creator>Listian, S. A.</dc:creator>
<dc:creator>Hüttel, S. V.</dc:creator>
<dc:creator>Müller, D.</dc:creator>
<dc:creator>Schäfer, D. G.</dc:creator>
<dc:creator>Vormittag, S.</dc:creator>
<dc:creator>Hilbi, H.</dc:creator>
<dc:creator>Maniak, M.</dc:creator>
<dc:creator>Gutierrez, M.</dc:creator>
<dc:creator>Barisch, C.</dc:creator>
<dc:date>2023-04-19</dc:date>
<dc:identifier>doi:10.1101/2023.04.17.537276</dc:identifier>
<dc:title><![CDATA[ER-dependent membrane repair of mycobacteria-induced vacuole damage]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.04.18.536394v1?rss=1">
<title>
<![CDATA[
CRISPR-Cas9 mediated endogenous utrophin upregulation improves Duchenne Muscular Dystrophy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.04.18.536394v1?rss=1"
</link>
<description><![CDATA[
Duchenne muscular dystrophy (DMD) is a lethal neuromuscular disorder caused by loss of dystrophin. Upregulation of utrophin (UTRN), a dystrophin paralogue, is a promising therapeutic avenue. Here, we present a CRISPR-Cas9-mediated strategy to increase utrophin expression by disrupting microRNA (miR) binding sites (BS). Using a Cas9/gRNA ribonucleoprotein (RNP) complex we disrupted several miR BS in DMD myoblasts and selected the Let-7c BS has crucial for UTRN repression. Interestingly, Cas9/gRNA indels were as efficient as the complete removal of Let-7c BS in upregulating UTRN expression, without any major off-targets. In three-dimensional human DMD cultures, Cas9/gRNA-mediated editing resulted in significant utrophin upregulation and functional improvements of calcium dysregulation and muscle contraction. Finally, Let-7c BS disruption in mdx animals by systemic rAAVs mediated delivery of Cas9 and gRNA resulted in utrophin upregulation and amelioration of the muscle histopathological phenotype. These findings provide the foundations for a universal (mutation-independent) gene editing therapeutic strategy for DMD.

One Sentence SummaryCRISPR-Cas9 has the potential to upregulate utrophin to treat all DMD patients.
]]></description>
<dc:creator>Guiraud, S.</dc:creator>
<dc:creator>Dastidar, S.</dc:creator>
<dc:creator>Mazed, F.</dc:creator>
<dc:creator>Amor, F.</dc:creator>
<dc:creator>Ralu, M.</dc:creator>
<dc:creator>de Cian, A.</dc:creator>
<dc:creator>Richard, I.</dc:creator>
<dc:creator>Ronzitti, G.</dc:creator>
<dc:creator>Tedesco, F. S.</dc:creator>
<dc:creator>Amendola, M.</dc:creator>
<dc:date>2023-04-19</dc:date>
<dc:identifier>doi:10.1101/2023.04.18.536394</dc:identifier>
<dc:title><![CDATA[CRISPR-Cas9 mediated endogenous utrophin upregulation improves Duchenne Muscular Dystrophy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.04.19.537498v1?rss=1">
<title>
<![CDATA[
Altered neutrophil extracellular traps in response to Mycobacterium tuberculosis in persons living with HIV with no previous TB and negative TST and IGRA 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.04.19.537498v1?rss=1"
</link>
<description><![CDATA[
Persons living with HIV (PLWH) have an increased risk for tuberculosis (TB). After prolonged and repeated exposure, some PLWH never develop TB and test persistently negative in tests of immune sensitization tuberculin skin test (TST) and interferon gamma release assays (IGRA) for Mycobacterium tuberculosis (Mtb). This group has been identified and defined as HIV+ persistently TB, tuberculin and IGRA negative (HITTIN). To investigate potential innate mechanisms unique to individuals with the HITTIN phenotype we compared their neutrophil Mtb infection response to that of PLWH, with no TB history, but who test persistently IGRA positive, and tuberculin positive (HIT). Neutrophil samples from 17 HITTIN (PMNHITTIN) and 11 HIT (PMNHIT) were isolated and infected with Mtb H37Rv for 1h and 6h. RNA was extracted and used for RNAseq analysis. At 1h of Mtb infection, PMNHITTIN displayed 151 significantly upregulated and 40 significantly downregulated differentially expressed genes (DEGs) and PMNHIT 98 significantly upregulated and 11 significantly downregulated DEGs. At the 6h timepoint, PMNHITTIN displayed 3106 significantly upregulated and 3548 significantly downregulated DEGs while PMNHIT had 3816 significantly up- and 3794 significantly downregulated DEGs. There was no significant differential transcriptional response at 1h between infected PMNHITTIN and PMNHIT. However, when contrasting the log2FC 6h infection response to Mtb from PMNHITTIN against PMNHIT, 2285 genes showed significant differential response between the two groups. Apoptosis and NETosis were key pathways linked to the enrichment of genes in PMNHITTIN when contrasted to PMNHIT after 6h infection with Mtb. Fluorescence microscopy revealed relatively lower neutrophil extracellular trap formation and cell loss in PMNHITTIN compared to PMNHIT, showing that PMNHITTIN have a distinct response to Mtb.
]]></description>
<dc:creator>Kroon, E. E.</dc:creator>
<dc:creator>Correa de Macedo, W.</dc:creator>
<dc:creator>Evans, R.</dc:creator>
<dc:creator>Seeger, A.</dc:creator>
<dc:creator>Engelbrecht, L.</dc:creator>
<dc:creator>Kriel, J.</dc:creator>
<dc:creator>Loos, B.</dc:creator>
<dc:creator>Okugbeni, N.</dc:creator>
<dc:creator>Orlova, M. O.</dc:creator>
<dc:creator>Cassart, P.</dc:creator>
<dc:creator>Kinnear, C.</dc:creator>
<dc:creator>Tromp, G.</dc:creator>
<dc:creator>Moller, M.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Coussens, A. K.</dc:creator>
<dc:date>2023-04-21</dc:date>
<dc:identifier>doi:10.1101/2023.04.19.537498</dc:identifier>
<dc:title><![CDATA[Altered neutrophil extracellular traps in response to Mycobacterium tuberculosis in persons living with HIV with no previous TB and negative TST and IGRA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.04.19.537022v1?rss=1">
<title>
<![CDATA[
Circulating neutrophils from patients with early breast cancer have distinct subtype-dependent phenotypes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.04.19.537022v1?rss=1"
</link>
<description><![CDATA[
PurposeA high number of circulating neutrophils is a poor prognostic factor for breast cancer, where evidence of bone marrow cancer-dependent priming is found. However, how early this priming is detectable remains unclear.

Patients and MethodsHere, we investigate changes in circulating neutrophils from newly diagnosed breast cancer patients before any therapeutic interventions. To do this, we assessed their lifespan and their broader intracellular kinase network activation states by using the Pamgene Kinome assay which measures the activity of neutrophil kinases.

ResultsWe found sub-type specific L-selectin (CD62L) changes in circulating neutrophils as well as perturbations in their overall global kinase activity. Strikingly, breast cancer patients of different subtypes (HR+, HER2+, triple negative) exhibited distinct neutrophil kinase activity patterns indicating that quantifiable perturbations can be detected in circulating neutrophils from early breast cancer patients, that are sensitive to both hormonal and HER-2 status. We also detected an increase in neutrophils lifespan in cancer patients, independently of tumour subtype.

ConclusionsOur results suggest that the tumour-specific kinase activation patterns in circulating neutrophils may be used in conjunction with other markers to identify patients with cancer from those harbouring only benign lesions of the breast. Given the important role neutrophil in breast cancer progression, the significance of this sub-type of specific priming warrants further investigation.

Clinical RelevanceThe current study aims to investigate cancer-specific changes in circulating neutrophils in patients with newly diagnosed early breast cancer before any therapeutic intervention. We found L-selectin (CD62L) changes in circulating neutrophils from patients with early-stage breast cancer compared to healthy volunteers, which is an indication of an early phenotypical change. Moreover, these changes in CD62L were dependent on the breast cancer sub-type, showing opposing trends according to the hormonal receptor status of the tumour. Importantly, this subtype dependent phenotypic alteration was reflected in broader intracellular signalling perturbation when measuring intracellular kinase activity. Moreover, those cancer perturbed neutrophils, show expanded life span when cultured ex vivo, suggesting an alteration in their physiologic state. The tumour-specific kinase activation patterns in circulating neutrophils may be useful in conjunction with other markers to distinguish patients with cancer from those with benign lesions of the breast.
]]></description>
<dc:creator>Ramessur, A.</dc:creator>
<dc:creator>Ambasager, B.</dc:creator>
<dc:creator>Aramburu, I. V.</dc:creator>
<dc:creator>Peakman, F.</dc:creator>
<dc:creator>Gleason, K.</dc:creator>
<dc:creator>Lehmann, C.</dc:creator>
<dc:creator>Papayannopoulos, V.</dc:creator>
<dc:creator>Coombes, C.</dc:creator>
<dc:creator>Malanchi, I.</dc:creator>
<dc:date>2023-04-21</dc:date>
<dc:identifier>doi:10.1101/2023.04.19.537022</dc:identifier>
<dc:title><![CDATA[Circulating neutrophils from patients with early breast cancer have distinct subtype-dependent phenotypes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.04.21.537779v1?rss=1">
<title>
<![CDATA[
Mapping host-microbe transcriptional interactions by dual perturb-seq 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.04.21.537779v1?rss=1"
</link>
<description><![CDATA[
Intracellular pathogens and other endosymbionts reprogram host cell transcription to suppress immune responses and recalibrate biosynthetic pathways. This reprogramming is critical in determining the outcome of infection or colonisation. Here, we combine pooled CRISPR knockout screening with dual host-microbe single-cell RNA-sequencing to identify the molecular mediators of these transcriptional interactions, a method we term dual perturb-seq. Applying dual perturb-seq to the intracellular pathogen Toxoplasma gondii, we are able to identify previously uncharacterised effector proteins and directly infer their function from the transcriptomic data. We show that TgGRA59 contributes to the export of other effector proteins from the parasite into the host cell and identify a novel effector, TgSOS1, that is necessary for sustained host STAT6 signalling and thereby contributes to parasite immune evasion and persistence. Together, this work demonstrates a novel tool that can be broadly adapted to interrogate host-microbe transcriptional interactions and reveal mechanisms of infection and immune evasion.
]]></description>
<dc:creator>Butterworth, S.</dc:creator>
<dc:creator>Kordova, K.</dc:creator>
<dc:creator>Chandrasekaran, S.</dc:creator>
<dc:creator>Thomas, K. K.</dc:creator>
<dc:creator>Torelli, F.</dc:creator>
<dc:creator>Lockyer, E. J.</dc:creator>
<dc:creator>Edwards, A.</dc:creator>
<dc:creator>Goldstone, R.</dc:creator>
<dc:creator>Koshy, A. A.</dc:creator>
<dc:creator>Treeck, M.</dc:creator>
<dc:date>2023-04-21</dc:date>
<dc:identifier>doi:10.1101/2023.04.21.537779</dc:identifier>
<dc:title><![CDATA[Mapping host-microbe transcriptional interactions by dual perturb-seq]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.04.24.538049v1?rss=1">
<title>
<![CDATA[
Discovery and characterization of a specific inhibitor of serine-threonine kinase cyclin dependent kinase-like 5 (CDKL5) demonstrates role in hippocampal CA1 physiology 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.04.24.538049v1?rss=1"
</link>
<description><![CDATA[
Pathological loss-of-function mutations in cyclin-dependent kinase-like 5 (CDKL5) cause CDKL5 deficiency disorder (CDD), a rare and severe neurodevelopmental disorder associated with severe and medically refractory early-life epilepsy, motor, cognitive, visual and autonomic disturbances in the absence of any structural brain pathology. Analysis of genetic variants in CDD have indicated that CDKL5 kinase function is central to disease pathology. CDKL5 encodes a serine-threonine kinase with significant homology to GSK3{beta}, which has also been linked to synaptic function. Further, Cdkl5 knock-out rodents have increased GSK3{beta} activity and often increased long-term potentiation (LTP). Thus, development of a specific CDKL5 inhibitor must be careful to exclude cross-talk with GSK3{beta} activity. We synthesized and characterized specific, high-affinity inhibitors of CDKL5 that do not have detectable activity for GSK3{beta}. These compounds are very soluble in water but blood-brain barrier penetration is low. In rat hippocampal brain slices, acute inhibition of CDKL5 selectively reduces post-synaptic function of AMPA-type glutamate receptors in a dose-dependent manner. Acute inhibition of CDKL5 reduces hippocampal LTP. These studies provide new tools and insights into the role of CDKL5 as a newly appreciated, key kinase necessary for synaptic plasticity. Comparisons to rodent knock-out studies suggest that compensatory changes have limited the understanding of the roles of CDKL5 in synaptic physiology, plasticity and human neuropathology.
]]></description>
<dc:creator>Castano, A.</dc:creator>
<dc:creator>Silvestre, M.</dc:creator>
<dc:creator>Wells, C. I.</dc:creator>
<dc:creator>Sanderson, J. L.</dc:creator>
<dc:creator>Ferrer, C. A.</dc:creator>
<dc:creator>Ong, H. W.</dc:creator>
<dc:creator>Lang, Y.</dc:creator>
<dc:creator>Richardson, W.</dc:creator>
<dc:creator>Silvaroli, J. A.</dc:creator>
<dc:creator>Bashore, F. M.</dc:creator>
<dc:creator>Smith, J. L.</dc:creator>
<dc:creator>Genereux, I. M.</dc:creator>
<dc:creator>Dempster, K.</dc:creator>
<dc:creator>Drewry, D. H.</dc:creator>
<dc:creator>Pabla, N. S.</dc:creator>
<dc:creator>Bullock, A. N.</dc:creator>
<dc:creator>Benke, T. A.</dc:creator>
<dc:creator>Ultanir, S.</dc:creator>
<dc:creator>Axtman, A. D.</dc:creator>
<dc:date>2023-04-24</dc:date>
<dc:identifier>doi:10.1101/2023.04.24.538049</dc:identifier>
<dc:title><![CDATA[Discovery and characterization of a specific inhibitor of serine-threonine kinase cyclin dependent kinase-like 5 (CDKL5) demonstrates role in hippocampal CA1 physiology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.04.27.538568v1?rss=1">
<title>
<![CDATA[
Abundant capped RNAs are derived from mRNA cleavage at 3'UTR G-Quadruplexes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.04.27.538568v1?rss=1"
</link>
<description><![CDATA[
The 3 untranslated region (3UTR) plays a crucial role in determining mRNA stability, localisation, translation and degradation. Cap analysis gene expression (CAGE), a method for the detection of capped 5 ends of mRNAs, additionally reveals a large number of apparently 5 capped RNAs derived from 3UTRs. Here we provide the first direct evidence that these 3UTR-derived RNAs are indeed capped and often more abundant than the corresponding full-length mRNAs. By using a combination of AGO2 enhanced individual nucleotide resolution UV crosslinking and immunoprecipitation (eiCLIP) and CAGE following siRNA knockdowns, we find that these 3UTR-derived RNAs likely originate from AGO2-mediated cleavage, and most often occur at locations with potential to form RNA-G-quadruplexes and are enriched by RNA-binding protein UPF1. High-resolution imaging and long-read sequencing analysis validates several 3UTR-derived RNAs, demonstrates their abundance and shows that they tend not to co-localise with the parental mRNAs. We also find that production of 3UTR-derived RNA could explain the previously reported role of a 3UTR G-quadruplex in regulating the production of APP protein. Taken together, we provide new insights into the origin and abundance of 3UTR-derived RNAs, show the utility of CAGE-seq for their quantitative detection, and provide a rich dataset for exploring new biology of a poorly understood new class of RNAs.

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=73 SRC="FIGDIR/small/538568v3_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Haberman, N.</dc:creator>
<dc:creator>Digby, H.</dc:creator>
<dc:creator>Faraway, R.</dc:creator>
<dc:creator>Cheung, R.</dc:creator>
<dc:creator>Jobbins, A. M.</dc:creator>
<dc:creator>Parr, C.</dc:creator>
<dc:creator>Yasuzawa, K.</dc:creator>
<dc:creator>Kasukawa, T.</dc:creator>
<dc:creator>Yip, C. W.</dc:creator>
<dc:creator>Kato, M.</dc:creator>
<dc:creator>Takahashi, H.</dc:creator>
<dc:creator>Carninci, P.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Sibley, C. R.</dc:creator>
<dc:creator>Martinez-sanchez, A.</dc:creator>
<dc:creator>Lenhard, B.</dc:creator>
<dc:creator>Vernia, S.</dc:creator>
<dc:date>2023-04-27</dc:date>
<dc:identifier>doi:10.1101/2023.04.27.538568</dc:identifier>
<dc:title><![CDATA[Abundant capped RNAs are derived from mRNA cleavage at 3'UTR G-Quadruplexes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.04.27.538529v1?rss=1">
<title>
<![CDATA[
Rare SH2B3 coding variants identified in lupus patients impair B cell tolerance and predispose to autoimmunity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.04.27.538529v1?rss=1"
</link>
<description><![CDATA[
Systemic lupus erythematosus (SLE) is a heterogeneous autoimmune disease, with a clear genetic component. While most SLE patients carry rare gene variants in lupus risk genes, little is known about their contribution to disease pathogenesis. Amongst them, SH2B3 - a negative regulator of cytokine and growth factor receptor signaling - harbors rare coding variants in over 5% of SLE patients. Here we show that unlike the variant found exclusively in healthy controls, most SH2B3 rare variants found in lupus patients are predominantly hypomorphic alleles. Generation of two mouse lines carrying variants orthologous to those found in patients revealed SH2B3 is important to limit the numbers of immature and transitional B cells. Furthermore, hypomorphic SH2B3 was shown to impair negative selection of immature/transitional self-reactive B cells and accelerate autoimmunity in sensitized mice, at least in part due to increased IL-4R signaling and BAFF-R expression. This work identifies a previously unappreciated role for SH2B3 in human B cell tolerance and lupus risk.

SummaryZhang et al. reveal a role for hypomorphic SH2B3 in lupus risk. The study shows rare and damaging variants identified in lupus patients enable breach of B cell immune tolerance checkpoints and suggests involvement for dysregulated IL-4R signaling and BAFF-R expression.
]]></description>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Morris, R.</dc:creator>
<dc:creator>Lorenzo, A. M. D.</dc:creator>
<dc:creator>Meng, X.</dc:creator>
<dc:creator>Kershaw, N. J.</dc:creator>
<dc:creator>Kiridena, P.</dc:creator>
<dc:creator>Brown, G. J.</dc:creator>
<dc:creator>Burgio, G.</dc:creator>
<dc:creator>Cappello, J. Y.</dc:creator>
<dc:creator>Shen, Q.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Turnbull, C. M.</dc:creator>
<dc:creator>Lea-Henry, T.</dc:creator>
<dc:creator>Stanley, M.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Ballard, F.</dc:creator>
<dc:creator>Chuah, A.</dc:creator>
<dc:creator>Lee, J. C.</dc:creator>
<dc:creator>Hatch, A.-M.</dc:creator>
<dc:creator>Headley, A. P.</dc:creator>
<dc:creator>Trnka, P.</dc:creator>
<dc:creator>Mallon, D.</dc:creator>
<dc:creator>Fletcher, J. T.</dc:creator>
<dc:creator>Walters, G. D.</dc:creator>
<dc:creator>Sestan, M.</dc:creator>
<dc:creator>Jelusic, M.</dc:creator>
<dc:creator>Cook, M. C.</dc:creator>
<dc:creator>Athanasopoulos, V.</dc:creator>
<dc:creator>Fulcher, D. A.</dc:creator>
<dc:creator>Babon, J. J.</dc:creator>
<dc:creator>Vinuesa, C. G.</dc:creator>
<dc:creator>Ellyard, J. I.</dc:creator>
<dc:date>2023-04-29</dc:date>
<dc:identifier>doi:10.1101/2023.04.27.538529</dc:identifier>
<dc:title><![CDATA[Rare SH2B3 coding variants identified in lupus patients impair B cell tolerance and predispose to autoimmunity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.05.02.539063v1?rss=1">
<title>
<![CDATA[
An image segmentation method based on the spatial correlation coefficient of Local Moran's I 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.05.02.539063v1?rss=1"
</link>
<description><![CDATA[
Unsupervised segmentation in biological and non-biological images is only partially resolved. Segmentation either requires arbitrary thresholds or large teaching datasets. Here we propose a spatial autocorrelation method based on Local Morans I coefficient to differentiate signal, background and noise in any type of image. The method, originally described for geoinformatics, does not require a predefined intensity threshold or teaching algorithm for image segmentation and allows quantitative comparison of samples obtained in different conditions. It utilizes relative intensity as well as spatial information of neighboring elements to select spatially contiguous groups of pixels. We demonstrate that Morans method outperforms threshold-based method (TBM) in both artificially generated as well as in natural images especially when background noise is substantial. This superior performance can be attributed to the exclusion of false positive pixels resulting from isolated, high intensity pixels in high noise conditions. To test the methods power in real situation we used high power confocal images of the somatosensory thalamus immunostained for Kv4.2 and Kv4.3 (A-type) voltage gated potassium channels. Morans method identified high intensity Kv4.2 and Kv4.3 ion channel clusters in the thalamic neuropil. Spatial distribution of these clusters displayed strong correlation with large sensory axon terminals of subcortical origin. The unique association of the special presynaptic terminals and a postsynaptic voltage gated ion channel cluster was confirmed with electron microscopy. These data demonstrate that Morans method is a rapid, simple image segmentation method optimal for variable and high nose conditions.
]]></description>
<dc:creator>David, C.</dc:creator>
<dc:creator>Giber, K.</dc:creator>
<dc:creator>Kerti-Szigeti, K.</dc:creator>
<dc:creator>Kollo, M.</dc:creator>
<dc:creator>Nusser, Z.</dc:creator>
<dc:creator>Acsady, L.</dc:creator>
<dc:date>2023-05-02</dc:date>
<dc:identifier>doi:10.1101/2023.05.02.539063</dc:identifier>
<dc:title><![CDATA[An image segmentation method based on the spatial correlation coefficient of Local Moran's I]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.05.05.539522v1?rss=1">
<title>
<![CDATA[
A disease-associated gene desert orchestrates macrophage inflammatory responses via ETS2 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.05.05.539522v1?rss=1"
</link>
<description><![CDATA[
Increasing global rates of autoimmune and inflammatory disease present a burgeoning threat to human health1. This is compounded by the limited efficacy of available treatments1 and high failure rates during drug development2 - underscoring an urgent need to better understand disease mechanisms. Here we show how genetics could address this challenge. By investigating an intergenic haplotype on chr21q22, independently linked to inflammatory bowel disease (IBD), ankylosing spondylitis, primary sclerosing cholangitis and Takayasus arteritis3-6, we discover that the causal gene, ETS2, is a master regulator of inflammatory responses in human macrophages and delineate how the risk haplotype increases ETS2 expression. Genes regulated by ETS2 were prominently expressed in affected tissues from chr21q22-associated diseases and more enriched for IBD GWAS hits than almost all previously described pathways. Overexpressing ETS2 in resting macrophages produced an activated effector state that phenocopied intestinal macrophages from IBD7, with upregulation of multiple drug targets including TNF and IL-23. Using a database of cellular signatures8, we identify drugs that could modulate this pathway and validate the potent anti-inflammatory activity of one class of small molecules in vitro and ex vivo. Together, this highlights the potential for common genetic associations to improve both the understanding and treatment of human disease.
]]></description>
<dc:creator>Stankey, C. T.</dc:creator>
<dc:creator>Bourges, C.</dc:creator>
<dc:creator>Turner-Stokes, T.</dc:creator>
<dc:creator>Piedade, A. P.</dc:creator>
<dc:creator>Palmer-Jones, C.</dc:creator>
<dc:creator>Papa, I.</dc:creator>
<dc:creator>Silva dos Santos, M.</dc:creator>
<dc:creator>Randzavola, L. O.</dc:creator>
<dc:creator>Speidel, L.</dc:creator>
<dc:creator>Parkes, E. C.</dc:creator>
<dc:creator>Edwards, W.</dc:creator>
<dc:creator>Rochford, A. P.</dc:creator>
<dc:creator>Murray, C.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Skoglund, P.</dc:creator>
<dc:creator>Wallace, C.</dc:creator>
<dc:creator>Cader, M. Z.</dc:creator>
<dc:creator>Thomas, D. C.</dc:creator>
<dc:creator>Lee, J. C.</dc:creator>
<dc:date>2023-05-07</dc:date>
<dc:identifier>doi:10.1101/2023.05.05.539522</dc:identifier>
<dc:title><![CDATA[A disease-associated gene desert orchestrates macrophage inflammatory responses via ETS2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.05.11.540445v1?rss=1">
<title>
<![CDATA[
CLEM-Reg: An automated point cloud based registration algorithm for correlative light and volume electron microscopy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.05.11.540445v1?rss=1"
</link>
<description><![CDATA[
Correlative light and volume electron microscopy (vCLEM) is a powerful imaging technique that enables the visualisation of fluorescently labelled proteins within their ultrastructural context on a subcellular level. Currently, expert microscopists align vCLEM acquisitions using time-consuming and subjective manual methods. This paper presents CLEM-Reg, an algorithm that automates the 3D alignment of vCLEM datasets by leveraging probabilistic point cloud registration techniques. These point clouds are derived from segmentations of common structures in each modality, created by state-of-the-art open-source methods, with the option to leverage alternative tools from other plugins or platforms. CLEM-Reg drastically reduces the time required to register vCLEM datasets to a few minutes and achieves correlation of fluorescent signal to sub-micron target structures in EM on three newly acquired vCLEM benchmark datasets (fluorescence microscopy combined with FIB-SEM or SBF-SEM). CLEM-Reg was then used to automatically obtain vCLEM overlays to unambiguously identify TGN46-positive transport carriers involved in the trafficking of proteins between the trans-Golgi network and plasma membrane. The datasets are available in the EMPIAR and BioStudies public image archives for reuse in testing and developing multimodal registration algorithms by the wider community. A napari plugin integrating the algorithm is also provided to aid end-user adoption.
]]></description>
<dc:creator>Krentzel, D.</dc:creator>
<dc:creator>Elphick, M.</dc:creator>
<dc:creator>Domart, M.-C.</dc:creator>
<dc:creator>Peddie, C. J.</dc:creator>
<dc:creator>Laine, R. F.</dc:creator>
<dc:creator>Henriques, R.</dc:creator>
<dc:creator>Collinson, L. M.</dc:creator>
<dc:creator>Jones, M. L.</dc:creator>
<dc:date>2023-05-12</dc:date>
<dc:identifier>doi:10.1101/2023.05.11.540445</dc:identifier>
<dc:title><![CDATA[CLEM-Reg: An automated point cloud based registration algorithm for correlative light and volume electron microscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.05.17.541126v1?rss=1">
<title>
<![CDATA[
mRNA display reveals a class of high-affinity bromodomain-binding motifs that are not found in the human proteome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.05.17.541126v1?rss=1"
</link>
<description><![CDATA[
Bromodomains regulate gene expression by recognizing protein motifs containing acetyllysine. Although originally characterized as histone-binding proteins, it has since become clear that these domains interact with other acetylated proteins, perhaps most prominently transcription factors. The likely transient nature and low stoichiometry of such modifications, however, has made it challenging to fully define the interactome of any given bromodomain. To begin to address this knowledge gap in an unbiased manner, we carried out mRNA display screens against a bromodomain - the N-terminal bromodomain of BRD3 - using peptide libraries that contained either one or two acetyllysine residues. We discovered peptides with very strong consensus sequences and with affinities that are significantly higher than typical bromodomain-peptide interactions. X-ray crystal structures also revealed modes of binding that have not been seen with natural ligands. Intriguingly, however, our selected sequences are not found in the human proteome, perhaps suggesting that strong binders to bromodomains might have been selected against.
]]></description>
<dc:creator>Low, J. K.</dc:creator>
<dc:creator>Patel, K.</dc:creator>
<dc:creator>Solomon, P.</dc:creator>
<dc:creator>Norman, A.</dc:creator>
<dc:creator>Pachl, P.</dc:creator>
<dc:creator>Jones, N.</dc:creator>
<dc:creator>Payne, R. J.</dc:creator>
<dc:creator>Passioura, T.</dc:creator>
<dc:creator>Suga, H.</dc:creator>
<dc:creator>Walport, L. J.</dc:creator>
<dc:creator>Mackay, J.</dc:creator>
<dc:date>2023-05-17</dc:date>
<dc:identifier>doi:10.1101/2023.05.17.541126</dc:identifier>
<dc:title><![CDATA[mRNA display reveals a class of high-affinity bromodomain-binding motifs that are not found in the human proteome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.05.18.541344v1?rss=1">
<title>
<![CDATA[
Antigenic mapping of the hemagglutinin of the H9 subtype influenza A viruses using sera from Japanese quail (Coturnix c. japonica). 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.05.18.541344v1?rss=1"
</link>
<description><![CDATA[
Influenza A viruses (FLUAV) of the H9N2 subtype are zoonotic pathogens that cause significant economic damage to the poultry industry. Vaccination to prevent and control H9N2 infections in poultry is widely employed in the Middle East and Asia. We used phylogenetics and antigenic analysis to study the antigenic properties of the H9 hemagglutinin (HA) using sera produced in Japanese quail (Coturnix c. japonica). Consensus HA1 sequences were generated to capture antigenic diversity among isolates. We constructed chimeric H9N2 viruses containing the HA1 of each consensus sequence on a constant isogenic backbone. The resulting viruses were used to generate antisera from quail, a common and significant minor poultry species whose anti-HA response profiles remain poorly defined. Antigenic maps were generated by plotting the cross-hemagglutination inhibition (HI) data from the panel of quail sera against the chimeric constructs and 51 H9 field isolates. The chimeric antigens were divided into four different antigenic profiles (cyan, blue, orange, and red). Site-directed mutagenesis analysis showed 9 amino acid positions of antigenic relevance. Substitutions at amino acid positions 149, 150, and 180 (H9 HA numbering) had relatively significant impact on HI activity using quail sera. Substitutions E180A and R131K/E180A led to the most significant antigenic change transitions. This study provides insights into the antigenic profile of H9 FLUAVs, with important implications for understanding antigenic drift and improving vaccine development for use in minor poultry species.

IMPORTANCEDetermining the relevant amino acids involved in antigenic drift on the surface protein hemagglutinin (HA) is critical to understand influenza virus evolution and efficient assessment of vaccine strains relative to current circulating strains. We used antigenic cartography to generate an antigenic map of the H9 HA using sera produced in one of the most relevant minor poultry species, Japanese quail. Key antigenic positions were identified and tested to confirm their impact on the antigenic profile. This work provides a better understanding of the antigenic diversity of the H9 HA as it relates to reactivity to quail sera and will facilitate a rational approach for selecting more efficacious vaccines against poultry-origin H9 influenza viruses in minor poultry species.
]]></description>
<dc:creator>Carnaccini, S.</dc:creator>
<dc:creator>Caceres, C. J.</dc:creator>
<dc:creator>Gay, L. C.</dc:creator>
<dc:creator>Ferreri, L.</dc:creator>
<dc:creator>Skepner, E.</dc:creator>
<dc:creator>Burke, D. F.</dc:creator>
<dc:creator>Brown, I. H.</dc:creator>
<dc:creator>Geiger, G.</dc:creator>
<dc:creator>Obadan, A.</dc:creator>
<dc:creator>Rajao, D. S.</dc:creator>
<dc:creator>Lewis, N. S.</dc:creator>
<dc:creator>Perez, D. R.</dc:creator>
<dc:date>2023-05-19</dc:date>
<dc:identifier>doi:10.1101/2023.05.18.541344</dc:identifier>
<dc:title><![CDATA[Antigenic mapping of the hemagglutinin of the H9 subtype influenza A viruses using sera from Japanese quail (Coturnix c. japonica).]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.05.17.538815v1?rss=1">
<title>
<![CDATA[
Drosophila AHR limits tumor growth and stem cell proliferation in the intestine 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.05.17.538815v1?rss=1"
</link>
<description><![CDATA[
The aryl hydrocarbon receptor (AHR) plays important roles in intestinal homeostasis, limiting tumour growth and promoting differentiation in the intestinal epithelium. Spineless, the Drosophila homolog of AHR, has only been studied in the context of development but not in the adult intestine. Here, we show that spineless is upregulated in the adult intestinal epithelium after infection with Pseudomonas entomophila (P.e.). Spineless knockdown increased stem cell proliferation following infection-induced injury. Spineless overexpression limited intestinal stem cell proliferation and reduced survival after infection. In two tumour models, using either Notch RNAi or constitutively active Yorkie, Spineless suppressed tumour growth and doubled the lifespan of tumour-bearing flies. At the transcriptional level it reversed the gene expression changes induced in Yorkie tumours, counteracting cell proliferation and altered metabolism. These findings demonstrate a new role for Spineless in the adult Drosophila midgut and highlight the evolutionarily conserved functions of AHR/Spineless in the control of proliferation and differentiation of the intestinal epithelium.
]]></description>
<dc:creator>Tsai, M.</dc:creator>
<dc:creator>Sun, J.</dc:creator>
<dc:creator>Alexandre, C.</dc:creator>
<dc:creator>Shapiro, M.</dc:creator>
<dc:creator>Franchet, A.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Gould, A. P.</dc:creator>
<dc:creator>Vincent, J.-P.</dc:creator>
<dc:creator>Stockinger, B.</dc:creator>
<dc:creator>Diny, N. L.</dc:creator>
<dc:date>2023-05-19</dc:date>
<dc:identifier>doi:10.1101/2023.05.17.538815</dc:identifier>
<dc:title><![CDATA[Drosophila AHR limits tumor growth and stem cell proliferation in the intestine]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.05.19.541416v1?rss=1">
<title>
<![CDATA[
RPM: an open-source rotation platform for open- and closed-loop vestibular stimulation in head-fixed mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.05.19.541416v1?rss=1"
</link>
<description><![CDATA[
Head fixation allows the recording and presentation of controlled stimuli and is used to study neural processes underlying spatial navigation. However, it disrupts the head direction system because of the lack of vestibular stimulation.

To overcome this limitation, we developed a novel rotation platform which can be driven by the experimenter (open-loop) or by animal movement (closed-loop). The platform is modular, affordable, easy to build and open source. Additional modules presented here include cameras for monitoring eye movements, visual virtual reality and a micro-manipulator for positioning various probes for recording or optical interference.

We demonstrate the utility of the platform by recording eye movements and showing the robust activation of head-direction cells. This novel experimental apparatus combines the advantages of head fixation and intact vestibular activity in the horizontal plane. The open-loop mode can be used to study e.g. vestibular sensory representation and processing, while the closed-loop mode allows animals to navigate in rotational space, providing a better substrate for 2-D navigation in virtual environments. Documentation is available at (https://ranczlab.github.io/RPM/).
]]></description>
<dc:creator>Cano-Ferrer, X.</dc:creator>
<dc:creator>Tran-Van-Minh, A.</dc:creator>
<dc:creator>Rancz, E.</dc:creator>
<dc:date>2023-05-22</dc:date>
<dc:identifier>doi:10.1101/2023.05.19.541416</dc:identifier>
<dc:title><![CDATA[RPM: an open-source rotation platform for open- and closed-loop vestibular stimulation in head-fixed mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.05.18.541303v1?rss=1">
<title>
<![CDATA[
A chromatinized origin reduces ORC mobility and spatially constrains MCM 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.05.18.541303v1?rss=1"
</link>
<description><![CDATA[
Chromatin replication involves the assembly and activity of the replisome within the nucleosomal landscape. At the core of the replisome is the Mcm2-7 complex (MCM), which is loaded onto DNA after binding to the Origin Recognition Complex (ORC). In yeast, ORC is a dynamic protein that diffuses rapidly along DNA, unless halted by origin recognition sequences. However, less is known about the dynamics of ORC proteins in the presence of nucleosomes and attendant consequences for MCM loading. To address this, we harnessed an in vitro single-molecule approach to interrogate a chromatinized origin of replication. We find that ORC binds the origin of replication with similar efficiency independently of whether the origin is chromatinized, despite ORC mobility being reduced by the presence of nucleosomes. Recruitment of MCM also proceeds efficiently on a chromatinized origin, but subsequent movement of MCM away from the origin is severely constrained. These findings suggest that chromatinized origins in yeast are essential for the local retention of MCM, which may facilitate subsequent assembly of the replisome.
]]></description>
<dc:creator>Sanchez, H.</dc:creator>
<dc:creator>Liu, Z.</dc:creator>
<dc:creator>van Veen, E.</dc:creator>
<dc:creator>van Laar, T.</dc:creator>
<dc:creator>Solano, B.</dc:creator>
<dc:creator>Diffley, J. F. X.</dc:creator>
<dc:creator>Dekker, N. H.</dc:creator>
<dc:date>2023-05-22</dc:date>
<dc:identifier>doi:10.1101/2023.05.18.541303</dc:identifier>
<dc:title><![CDATA[A chromatinized origin reduces ORC mobility and spatially constrains MCM]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.05.23.541898v1?rss=1">
<title>
<![CDATA[
PfAP2-MRP DNA-binding protein is a master regulator of parasite pathogenesis during malaria parasite blood stages 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.05.23.541898v1?rss=1"
</link>
<description><![CDATA[
Malaria pathogenicity results from the parasites ability to invade, multiply within and then egress from the host red blood cell (RBC). Infected RBCs are remodeled, expressing antigenic variant proteins (such as PfEMP1, coded by the var gene family) for immune evasion and survival. These processes require the concerted actions of many proteins, but the molecular regulation is poorly understood. We have characterized an essential Plasmodium specific Apicomplexan AP2 (ApiAP2) transcription factor in Plasmodium falciparum (PfAP2-MRP; Master Regulator of Pathogenesis) during the intraerythrocytic developmental cycle (IDC). An inducible gene knockout approach showed that PfAP2-MRP is essential for development during the trophozoite stage, and critical for var gene regulation, merozoite development and parasite egress. ChIP-seq experiments performed at 16 hour post invasion (h.p.i.) and 40 h.p.i. matching the two peaks of PfAP2-MRP expression, demonstrate binding of PfAP2-MRP to the promoters of genes controlling trophozoite development and host cell remodeling at 16 h.p.i. and antigenic variation and pathogenicity at 40 h.p.i. Using single-cell RNA-seq and fluorescence-activated cell sorting, we show de-repression of most var genes in{Delta} pfap2-mrp parasites that express multiple PfEMP1 proteins on the surface of infected RBCs. In addition, the{Delta} pfap2-mrp parasites overexpress several early gametocyte marker genes at both 16 and 40 h.p.i., indicating a regulatory role in the sexual stage conversion. Using the Chromosomes Conformation Capture experiment (Hi-C), we demonstrate that deletion of PfAP2-MRP results in significant reduction of both intra-chromosomal and inter-chromosomal interactions in heterochromatin clusters. We conclude that PfAP2-MRP is a vital upstream transcriptional regulator controlling essential processes in two distinct developmental stages during the IDC that include parasite growth, chromatin structure and var gene expression.
]]></description>
<dc:creator>Pain, A.</dc:creator>
<dc:creator>Subudhi, A. K.</dc:creator>
<dc:creator>Green, J.</dc:creator>
<dc:creator>Satyam, R.</dc:creator>
<dc:creator>Lenz, T.</dc:creator>
<dc:creator>Salunke, R. P.</dc:creator>
<dc:creator>Shuaib, M.</dc:creator>
<dc:creator>Isaioglou, I.</dc:creator>
<dc:creator>Abel, S.</dc:creator>
<dc:creator>Gupta, M.</dc:creator>
<dc:creator>Esau, L.</dc:creator>
<dc:creator>Mourier, T.</dc:creator>
<dc:creator>Mfarrej, S.</dc:creator>
<dc:creator>Sivapurkar, R.</dc:creator>
<dc:creator>Stead, Z.</dc:creator>
<dc:creator>Ben-Rached, F.</dc:creator>
<dc:creator>Otswal, Y.</dc:creator>
<dc:creator>Sougrat, R.</dc:creator>
<dc:creator>Dada, A.</dc:creator>
<dc:creator>Kadamany, A. F.</dc:creator>
<dc:creator>Fishle, W.</dc:creator>
<dc:creator>Merzaban, J.</dc:creator>
<dc:creator>Knuepfer, E.</dc:creator>
<dc:creator>Ferguson, D. J. P.</dc:creator>
<dc:creator>Gupta, I.</dc:creator>
<dc:creator>Le Roch, K. G.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:date>2023-05-24</dc:date>
<dc:identifier>doi:10.1101/2023.05.23.541898</dc:identifier>
<dc:title><![CDATA[PfAP2-MRP DNA-binding protein is a master regulator of parasite pathogenesis during malaria parasite blood stages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.05.24.541920v1?rss=1">
<title>
<![CDATA[
Dichotomy of neutralizing antibody, B cell and T cell responses to SARS-CoV-2 vaccination and protection in healthy adults 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.05.24.541920v1?rss=1"
</link>
<description><![CDATA[
Heterogeneity in SARS-CoV-2 vaccine responses is not understood. Here, we identify four patterns of live-virus neutralizing antibody responses: individuals with hybrid immunity (with confirmed prior infection); rare individuals with low responses (paucity of S1-binding antibodies); and surprisingly, two further groups with distinct serological repertoires. One group - broad responders - neutralize a range of SARS-CoV-2 variants, whereas the other - narrow responders - neutralize fewer, less divergent variants. This heterogeneity does not correlate with Ancestral S1-binding antibody, rather the quality of the serological response. Furthermore, IgDlowCD27-CD137+ B cells and CCR6+ CD4+ T cells are enriched in broad responders before dose 3. Notably, broad responders have significantly longer infection-free time after their third dose. Understanding the control and persistence of these serological profiles could allow personalized approaches to enhance serological breadth after vaccination.
]]></description>
<dc:creator>Carr, E. J.</dc:creator>
<dc:creator>Townsley, H.</dc:creator>
<dc:creator>Wu, M. Y.</dc:creator>
<dc:creator>Wilkinson, K. A.</dc:creator>
<dc:creator>Hobson, P. S.</dc:creator>
<dc:creator>Levi, D.</dc:creator>
<dc:creator>Namjou, S.</dc:creator>
<dc:creator>Mears, H. V.</dc:creator>
<dc:creator>Hobbs, A.</dc:creator>
<dc:creator>Ragno, M.</dc:creator>
<dc:creator>Herman, L. S.</dc:creator>
<dc:creator>Harvey, R.</dc:creator>
<dc:creator>Bailey, C.</dc:creator>
<dc:creator>Fowler, A. S.</dc:creator>
<dc:creator>Hatipoglu, E.</dc:creator>
<dc:creator>Ngai, Y.</dc:creator>
<dc:creator>Clayton, B.</dc:creator>
<dc:creator>Miah, M.</dc:creator>
<dc:creator>Bawumia, P.</dc:creator>
<dc:creator>Miranda, M.</dc:creator>
<dc:creator>Smith, C.</dc:creator>
<dc:creator>Sawyer, C.</dc:creator>
<dc:creator>Kelly, G.</dc:creator>
<dc:creator>Mahalingasivam, V.</dc:creator>
<dc:creator>Zheng, B.</dc:creator>
<dc:creator>Evans, S. J.</dc:creator>
<dc:creator>Libri, V.</dc:creator>
<dc:creator>Riddell, A.</dc:creator>
<dc:creator>Nicod, J.</dc:creator>
<dc:creator>O'Reilly, N.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>Williams, B.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Kassiotis, G.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Beale, R. C.</dc:creator>
<dc:creator>Bauer, D. L.</dc:creator>
<dc:creator>Wall, E. C.</dc:creator>
<dc:date>2023-05-25</dc:date>
<dc:identifier>doi:10.1101/2023.05.24.541920</dc:identifier>
<dc:title><![CDATA[Dichotomy of neutralizing antibody, B cell and T cell responses to SARS-CoV-2 vaccination and protection in healthy adults]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.05.27.541573v1?rss=1">
<title>
<![CDATA[
Divergent evolution of sleep functions 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.05.27.541573v1?rss=1"
</link>
<description><![CDATA[
Most living organisms have evolved to synchronize their biological activities with the earths rotation, a daily regulation of biology and behaviour controlled by an evolutionary conserved molecular machinery known as the circadian clock. For most animals, circadian mechanisms are meant to maximize their exposure to positive activities (e.g.: social interactions, mating, feeding - generally during the day) and minimize their exposure to peril (e.g.: predation, weather, darkness - generally during the night1). On top of circadian regulation, some behaviours also feature a second layer of homeostatic control acting as a fail-safe to ensure important activities are not ignored. Sleep is one of these behaviours: largely controlled by the circadian clock for its baseline appearance, it is at the same time modulated by a - poorly understood - homeostatic regulator ensuring animals obey their species-specific amount of daily sleep2. An evolutionary conserved homeostatic control is often considered the main evidence for a core biological function of sleep beyond the trivial one (that is: keeping us out of trouble by limiting our energy expenditure and exposure to danger3,4) and it is hypothesized that sleep evolved around this mysterious basic biological function. Here we characterize sleep regulation in a group of seven species of the Drosophila genus at key evolutionary distances and representing a variety of ecological niche adaptations. We show that the spontaneous circadian-driven aspects of sleep are conserved among all species but the homeostatic regulation, unexpectedly, is not. We uncover differences in the behavioural, cell-biological and neuro-pharmacological aspects of sleep and suggest that, in Drosophilids, sleep primarily evolved to satisfy a circadian role, keeping animals immobile during dangerous hours of the day. The homeostatic functions of sleep evolved independently, in a species-specific fashion, and are not conserved.
]]></description>
<dc:creator>Joyce, M.</dc:creator>
<dc:creator>Falconio, F. A.</dc:creator>
<dc:creator>Blackhurst, L.</dc:creator>
<dc:creator>Prieto-Godino, L. L.</dc:creator>
<dc:creator>French, A. S.</dc:creator>
<dc:creator>Gilestro, G. F.</dc:creator>
<dc:date>2023-05-28</dc:date>
<dc:identifier>doi:10.1101/2023.05.27.541573</dc:identifier>
<dc:title><![CDATA[Divergent evolution of sleep functions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.05.29.541316v1?rss=1">
<title>
<![CDATA[
Delay of human early development via in vitro diapause 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.05.29.541316v1?rss=1"
</link>
<description><![CDATA[
Many mammals can control the timing of gestation and birth by pausing embryonic development at the blastocyst stage. It is unknown whether the capacity to pause development is conserved, in general across mammals, and more specifically in humans. Activity of the growth regulating mTOR pathway governs developmental pausing in the mouse (1). Here we show a stage-specific capacity to delay the progression of human development via mTOR inhibition. In this context, human blastoids and pluripotent stem cells in naive and naive-like, but not primed, states can be induced to enter a dormant state, which is reversible at the functional and molecular level. Comparative analysis of mouse and human naive cells longitudinal response to mTORi revealed distinct temporal dynamics and metabolic requirements of dormancy in each species. Mouse and human blastocysts show similar tissue-specific patterns of mTOR pathway activity, suggesting that the mTOR pathway may be a conserved regulator of blastocyst development and timing in both species. Our results raise the possibility that the developmental timing of the human embryo may be controllable, with implications for reproductive therapies.
]]></description>
<dc:creator>Bulut-Karslioglu, A.</dc:creator>
<dc:creator>Iyer, D.</dc:creator>
<dc:creator>van der Weijden, V.</dc:creator>
<dc:creator>Heidari Khoei, H.</dc:creator>
<dc:creator>McCarthy, A.</dc:creator>
<dc:creator>Rayon, T.</dc:creator>
<dc:creator>Simon, C.</dc:creator>
<dc:creator>Dunkel, I.</dc:creator>
<dc:creator>Wamaitha, S.</dc:creator>
<dc:creator>Schulz, E.</dc:creator>
<dc:creator>Niakan, K.</dc:creator>
<dc:creator>Rivron, N. C.</dc:creator>
<dc:date>2023-05-29</dc:date>
<dc:identifier>doi:10.1101/2023.05.29.541316</dc:identifier>
<dc:title><![CDATA[Delay of human early development via in vitro diapause]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.05.31.539801v1?rss=1">
<title>
<![CDATA[
Single cell-guided prenatal derivation of primary epithelial organoids from the human amniotic and tracheal fluids. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.05.31.539801v1?rss=1"
</link>
<description><![CDATA[
Despite advances in prenatal diagnosis, it is still difficult to predict severity and outcomes of many congenital malformations. New patient-specific prenatal disease modelling may optimise personalised prediction. We and others have described the presence of mesenchymal stem cells in amniotic fluid (AFSC) that can generate induced pluripotent stem cells (iPSCs). The lengthy reprogramming processes, however, limits the ability to define individual phenotypes or plan prenatal treatment. Therefore, it would be advantageous if fetal stem cells could be obtained during pregnancy and expanded without reprogramming. Using single cell analysis, we characterised the cellular identities in amniotic fluid (AF) and identified viable epithelial stem/progenitor cells of fetal intestinal, renal and pulmonary origin. With relevance for prenatal disease modelling, these cells could be cultured to form clonal epithelial organoids manifesting small intestine, kidney and lung identity. To confirm this, we derived lung organoids from AF and tracheal fluid (TF) cells of Congenital Diaphragmatic Hernia (CDH) fetuses and found that they show differences to non-CDH controls and can recapitulate some pathological features of the disease. Amniotic Fluid Organoids (AFO) allow investigation of fetal epithelial tissues at clinically relevant developmental stages and may enable the development of therapeutic tools tailored to the fetus, as well as to predicting the effects of such therapies.
]]></description>
<dc:creator>Gerli, M. F. M.</dc:creator>
<dc:creator>Cala, G.</dc:creator>
<dc:creator>Beesley, M. A.</dc:creator>
<dc:creator>Sina, B.</dc:creator>
<dc:creator>Tullie, L.</dc:creator>
<dc:creator>Panariello, F.</dc:creator>
<dc:creator>Michielin, F.</dc:creator>
<dc:creator>Sun, K. Y.</dc:creator>
<dc:creator>Davidson, J. R.</dc:creator>
<dc:creator>Russo, F. M.</dc:creator>
<dc:creator>Jones, B. C.</dc:creator>
<dc:creator>Lee, D.</dc:creator>
<dc:creator>Savvidis, S.</dc:creator>
<dc:creator>Xenakis, T.</dc:creator>
<dc:creator>Simcock, I. C.</dc:creator>
<dc:creator>Straatman-Iwanowska, A. A.</dc:creator>
<dc:creator>Hirst, R. A.</dc:creator>
<dc:creator>David, A. L.</dc:creator>
<dc:creator>O'Callaghan, C.</dc:creator>
<dc:creator>Olivo, A.</dc:creator>
<dc:creator>Eaton, S.</dc:creator>
<dc:creator>Loukogeorgakis, S. P.</dc:creator>
<dc:creator>Cacchiarelli, D.</dc:creator>
<dc:creator>Deprest, J.</dc:creator>
<dc:creator>Li, V. S.</dc:creator>
<dc:creator>Giobbe, G. G.</dc:creator>
<dc:creator>De Coppi, P.</dc:creator>
<dc:date>2023-06-01</dc:date>
<dc:identifier>doi:10.1101/2023.05.31.539801</dc:identifier>
<dc:title><![CDATA[Single cell-guided prenatal derivation of primary epithelial organoids from the human amniotic and tracheal fluids.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.01.543217v1?rss=1">
<title>
<![CDATA[
Frictiotaxis underlies adhesion-independent durotaxis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.01.543217v1?rss=1"
</link>
<description><![CDATA[
Cells move directionally along gradients of substrate stiffness, a process called durotaxis. The current consensus is that durotaxis relies on cell-substrate focal adhesions to sense stiffness and transmit forces that drive directed motion. Therefore, focal adhesion-independent durotaxis is thought to be impossible. Here, we show that confined cells can perform durotaxis despite lacking strong or specific adhesions. This durotactic migration depends on asymmetric myosin distribution and actomyosin retrograde flow. We show that the mechanism of this adhesion-independent durotaxis is that stiffer substrates offer higher friction. We propose a physical model that predicts that non-adherent cells polarise and migrate towards regions of higher friction - a process that we call frictiotaxis. We demonstrate frictiotaxis in experiments by showing that cells migrate up a friction gradient even when stiffness is uniform. Our results broaden the potential of durotaxis to guide any cell that contacts a substrate and reveal a new mode of directed migration based on friction, with implications for immune and cancer cells, which commonly move with non-specific interactions.
]]></description>
<dc:creator>Shellard, A.</dc:creator>
<dc:creator>Hampshire, P.</dc:creator>
<dc:creator>Stillman, N.</dc:creator>
<dc:creator>Dix, C.</dc:creator>
<dc:creator>Thorogate, R.</dc:creator>
<dc:creator>Imbert, A.</dc:creator>
<dc:creator>Charras, G.</dc:creator>
<dc:creator>Alert, R.</dc:creator>
<dc:creator>Mayor, R.</dc:creator>
<dc:date>2023-06-01</dc:date>
<dc:identifier>doi:10.1101/2023.06.01.543217</dc:identifier>
<dc:title><![CDATA[Frictiotaxis underlies adhesion-independent durotaxis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.05.543701v1?rss=1">
<title>
<![CDATA[
Bridging Imaging Users to Imaging Analysis - A community survey 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.05.543701v1?rss=1"
</link>
<description><![CDATA[
The "Bridging Imaging Users to Imaging Analysis" survey was conducted in 2022 by the Center for Open Bioimage Analysis (COBA), Bioimaging North America (BINA), and the Royal Microscopical Society Data Analysis in Imaging Section (RMS DAIM) to understand the needs of the imaging community. Through multi-choice and open-ended questions, the survey inquired about demographics, image analysis experiences, future needs, and suggestions on the role of tool developers and users. Participants of the survey were from diverse roles and domains of the life and physical sciences. To our knowledge, this is the first attempt to survey cross-community to bridge knowledge gaps between physical and life sciences imaging. Survey results indicate that respondents overarching needs are documentation, detailed tutorials on the usage of image analysis tools, user-friendly intuitive software, and better solutions for segmentation, ideally in a format tailored to their specific use cases. The tool creators suggested the users familiarize themselves with the fundamentals of image analysis, provide constant feedback, and report the issues faced during image analysis while the users would like more documentation and an emphasis on tool friendliness. Regardless of the computational experience, there is a strong preference for  written tutorials to acquire knowledge on image analysis. We also observed that the interest in having  office hours to get an expert opinion on their image analysis methods has increased over the years. In addition, the community suggests the need for a common repository for the available image analysis tools and their applications. The opinions and suggestions of the community, released here in full, will help the image analysis tool creation and education communities to design and deliver the resources accordingly.
]]></description>
<dc:creator>Sivagurunathan, S.</dc:creator>
<dc:creator>Marcotti, S.</dc:creator>
<dc:creator>Nelson, C. J.</dc:creator>
<dc:creator>Jones, M. L.</dc:creator>
<dc:creator>Barry, D. J.</dc:creator>
<dc:creator>Slater, T. J. A.</dc:creator>
<dc:creator>Eliceiri, K. W.</dc:creator>
<dc:creator>Cimini, B. A.</dc:creator>
<dc:date>2023-06-05</dc:date>
<dc:identifier>doi:10.1101/2023.06.05.543701</dc:identifier>
<dc:title><![CDATA[Bridging Imaging Users to Imaging Analysis - A community survey]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.10.542698v1?rss=1">
<title>
<![CDATA[
Saturation Genome Editing Resolves the Functional Spectrum of Pathogenic VHL Alleles 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.10.542698v1?rss=1"
</link>
<description><![CDATA[
To maximize the impact of precision medicine approaches, it is critical to accurately identify genetic variants in cancer-associated genes with functional consequences. Yet, our knowledge of rare variants conferring clinically relevant phenotypes and the mechanisms through which they act remains highly limited. A tumor suppressor gene exemplifying the challenge of variant interpretation is VHL. VHL encodes an E3 ubiquitin ligase that regulates the cellular response to hypoxia. Germline pathogenic variants in VHL predispose patients to tumors including clear cell renal cell carcinoma (ccRCC) and pheochromocytoma, and somatic VHL mutations are frequently observed in sporadic renal cancer. Here, we optimize and apply Saturation Genome Editing (SGE) to assay nearly all possible single nucleotide variants (SNVs) across VHLs coding sequence. To delineate mechanisms, we quantify mRNA dosage effects over time and compare effects in isogenic cell lines. Function scores for 2,268 VHL SNVs identify a core set of pathogenic alleles driving ccRCC with perfect accuracy, inform differential risk across tumor types, and reveal novel mechanisms by which variants impact function. These results have immediate utility for classifying VHL variants encountered in both germline testing and tumor profiling and illustrate how precise functional measurements can resolve pleiotropic and dosage-dependent genotype-phenotype relationships across complete genes.
]]></description>
<dc:creator>Buckley, M.</dc:creator>
<dc:creator>Kajba, C. M.</dc:creator>
<dc:creator>Forrester, N. M.</dc:creator>
<dc:creator>Terwagne, C.</dc:creator>
<dc:creator>Sawyer, C.</dc:creator>
<dc:creator>Shepherd, S. T. C.</dc:creator>
<dc:creator>De Jonghe, J.</dc:creator>
<dc:creator>Dace, P.</dc:creator>
<dc:creator>Turajlic, S.</dc:creator>
<dc:creator>Findlay, G. M.</dc:creator>
<dc:date>2023-06-10</dc:date>
<dc:identifier>doi:10.1101/2023.06.10.542698</dc:identifier>
<dc:title><![CDATA[Saturation Genome Editing Resolves the Functional Spectrum of Pathogenic VHL Alleles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.10.544434v1?rss=1">
<title>
<![CDATA[
Evolution of chromosome arm aberrations in breast cancer through genetic network rewiring 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.10.544434v1?rss=1"
</link>
<description><![CDATA[
The basal breast cancer subtype is enriched for triple-negative breast cancer (TNBC) and displays consistent large chromosomal deletions. Here, we characterize the evolution and maintenance of chromosome 4p (chr4p) loss in basal breast cancer. TCGA data analysis showed recurrent deletion of chr4p in basal breast cancer. Phylogenetic analysis of a unique panel of 23 primary tumor/patient-derived xenograft basal breast cancers revealed early evolution of chr4p deletion. Mechanistically we show that Chr4p loss is associated with enhanced proliferation. Gene function studies identified an unknown gene, C4orf19, within chr4p, which suppressed proliferation when overexpressed and is a novel member of a PDCD10-GCKIII kinase module, we name as PGCA1. Genome-wide pooled overexpression screens using a barcoded library of human open reading frames, identified chromosomal regions, including chr4p, that suppress proliferation when overexpressed in a context-dependent manner implicating network interactions. Together this sheds light on the early emergence of complex aneuploid karyotypes involving chr4p and adaptive landscapes shaping breast cancer genomes.
]]></description>
<dc:creator>Kuzmin, E.</dc:creator>
<dc:creator>Baker, T.</dc:creator>
<dc:creator>Lesluyes, T.</dc:creator>
<dc:creator>Monlong, J.</dc:creator>
<dc:creator>Abe, K. T.</dc:creator>
<dc:creator>Coelho, P. P.</dc:creator>
<dc:creator>Schwartz, M.</dc:creator>
<dc:creator>Zou, D.</dc:creator>
<dc:creator>Morin, G.</dc:creator>
<dc:creator>Pacis, A.</dc:creator>
<dc:creator>Yang, Y.</dc:creator>
<dc:creator>Martinez, C.</dc:creator>
<dc:creator>Barber, J.</dc:creator>
<dc:creator>Kuasne, H.</dc:creator>
<dc:creator>Li, R.</dc:creator>
<dc:creator>Bourgey, M.</dc:creator>
<dc:creator>Fortier, A.-M.</dc:creator>
<dc:creator>Davison, P. G.</dc:creator>
<dc:creator>Omeroglu, A.</dc:creator>
<dc:creator>Guiot, M. C.</dc:creator>
<dc:creator>Morris, Q.</dc:creator>
<dc:creator>Kleinman, C. L.</dc:creator>
<dc:creator>Huang, S.</dc:creator>
<dc:creator>Gingras, A.-C.</dc:creator>
<dc:creator>Bourque, G.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>Park, M. M.</dc:creator>
<dc:date>2023-06-12</dc:date>
<dc:identifier>doi:10.1101/2023.06.10.544434</dc:identifier>
<dc:title><![CDATA[Evolution of chromosome arm aberrations in breast cancer through genetic network rewiring]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.14.545007v1?rss=1">
<title>
<![CDATA[
Beclin1 is essential for intestinal homeostasis. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.14.545007v1?rss=1"
</link>
<description><![CDATA[
BECLIN1 is a component of Class III phosphatidylinositol 3-kinase complexes that orchestrates autophagy initiation and endocytic trafficking. Here we show intestinal epithelium-specific BECLIN1 deletion in adult mice led to rapid fatal enteritis with compromised gut barrier integrity, highlighting its intrinsic critical role in gut maintenance. BECLIN1-deficient intestinal epithelial cells exhibited extensive apoptosis, impaired autophagy, and stressed endoplasmic reticulum and mitochondria. Remaining absorptive enterocytes and secretory cells displayed morphological abnormalities. Deletion of the autophagy regulator, ATG7, failed to elicit similar effects, suggesting novel autophagy-independent functions of BECLIN1 distinct from ATG7. Indeed, organoids derived from BECLIN1 KO mice showed E-cadherin mislocalisation providing a mechanism linking endocytic trafficking mediated by Beclin1 and loss of intestinal barrier integrity. Our findings establish an indispensable role of BECLIN1 in maintaining mammalian intestinal homeostasis and uncover its involvement in endocytic trafficking in this process. Hence, this study has significant implications for our understanding of intestinal pathophysiology.
]]></description>
<dc:creator>Tran, S.</dc:creator>
<dc:creator>Juliani, J.</dc:creator>
<dc:creator>Harris, T. J.</dc:creator>
<dc:creator>Evangelista, M.</dc:creator>
<dc:creator>Ratcliffe, J.</dc:creator>
<dc:creator>Ellis, S. L.</dc:creator>
<dc:creator>Baloyan, D.</dc:creator>
<dc:creator>Reehorst, C. M.</dc:creator>
<dc:creator>Nightingale, R.</dc:creator>
<dc:creator>Luk, I. Y.</dc:creator>
<dc:creator>Jenkins, L. J.</dc:creator>
<dc:creator>Ghilas, S.</dc:creator>
<dc:creator>Yakou, M. H.</dc:creator>
<dc:creator>Inguanti, C.</dc:creator>
<dc:creator>Johnson, C.</dc:creator>
<dc:creator>Buchert, M.</dc:creator>
<dc:creator>Lee, J. C.</dc:creator>
<dc:creator>De Cruz, P.</dc:creator>
<dc:creator>Duszyc, K.</dc:creator>
<dc:creator>Gleeson, P. A.</dc:creator>
<dc:creator>Kile, B. T.</dc:creator>
<dc:creator>Mielke, L. A.</dc:creator>
<dc:creator>Yap, A. S.</dc:creator>
<dc:creator>Mariadason, J.</dc:creator>
<dc:creator>Fairlie, W. D.</dc:creator>
<dc:creator>Lee, E. F.</dc:creator>
<dc:date>2023-06-15</dc:date>
<dc:identifier>doi:10.1101/2023.06.14.545007</dc:identifier>
<dc:title><![CDATA[Beclin1 is essential for intestinal homeostasis.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.15.545059v1?rss=1">
<title>
<![CDATA[
Gastruloid-derived Primordial Germ Cell-like Cells (Gld-PGCLCs) develop dynamically within integrated tissues 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.15.545059v1?rss=1"
</link>
<description><![CDATA[
Primordial Germ Cells (PGCs) are the early embryonic precursors of gametes - sperm and egg cells. PGC-like cells (PGCLCs) can currently be derived in vitro from pluripotent cells exposed to signalling cocktails and aggregated into large embryonic bodies, but these do not recapitulate the native embryonic environment during PGC formation. Here we show that mouse gastruloids, a three-dimensional in vitro model of gastrulation, contain a population of Gastruloid-derived PGC-like cells (Gld-PGCLCs) that resemble early PGCs in vivo. Importantly, the conserved organisation of mouse gastruloids leads to coordinated spatial and temporal localisation of Gld-PGCLCs relative to surrounding somatic cells, even in the absence of specific exogenous PGC-specific signalling or extraembryonic tissues. In gastruloids, self-organised interactions between cells and tissues, including the endodermal epithelium, enables the specification and subsequent maturation of a pool of Gld-PGCLCs. As such, mouse gastruloids represent a new source of PGCLCs in vitro and, due to their inherent co-development, serve as a novel model to study the dynamics of PGC development within integrated tissue environments.
]]></description>
<dc:creator>Cooke, C. B.</dc:creator>
<dc:creator>Barrington, C.</dc:creator>
<dc:creator>Baillie-Benson, P.</dc:creator>
<dc:creator>Nichols, J.</dc:creator>
<dc:creator>Moris, N.</dc:creator>
<dc:date>2023-06-15</dc:date>
<dc:identifier>doi:10.1101/2023.06.15.545059</dc:identifier>
<dc:title><![CDATA[Gastruloid-derived Primordial Germ Cell-like Cells (Gld-PGCLCs) develop dynamically within integrated tissues]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.15.545009v1?rss=1">
<title>
<![CDATA[
Impact of Streptococcus pneumoniae biosynthesis gene mutations on epithelial microinvasion and cellular responses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.15.545009v1?rss=1"
</link>
<description><![CDATA[
Nasopharyngeal colonisation by Streptococcus pneumoniae is characterised by bacterial adherence to epithelial cells, microinvasion and innate immune activation. Previously, we have shown two serotype 6B S. pneumoniae mutant strains affecting bacterial metabolism ({Delta}proABC/pia and{Delta} fhs/pia) colonise humans and mice, but in a murine disease model do not cause invasive infection. Here, we explore whether S. pneumoniae epithelial microinvasion and the induction of innate immune responses persist despite disease attenuation.

We show that under serum stress, these biosynthesis gene mutations had a broad but different impact on pneumococcal virulence gene expression, oxidative stress regulation, and purine and carbohydrate metabolism genes. However, although these mutations did not attenuate microinvasion in human challenge and epithelial models, there was less transmigration of Detroit 562 nasopharyngeal epithelial cells by the mutants compared to WT. Cellular reorganisation of primary human airway epithelium varied considerably between strains. Compared to WT, infection of Detroit 562 epithelial cells by the{Delta} fhs/piaA strain, but not the{Delta} proABC/piaA strain was less pro-inflammatory, induced less caspase 8 production, and were associated with increased pneumococcal hydrogen peroxide and reduced pneumolysin secretion.

These findings suggest that the observed differences in microinvasion and the epithelial response were driven by the differential expression of multiple bacterial virulence and metabolic pathways, rather than single genes or pathways of genes. These data highlight the complex impact of single gene mutations on bacterial virulence and suggest that the virulence determinants of pneumococcal epithelial colonisation, microinvasion and innate immunity are not necessarily directly linked to disease.

Author SummaryStreptococcus pneumoniae (the pneumococcus) commonly colonises the back of the human nose, and is a leading cause of pneumonia, meningitis, and sepsis. During colonisation, the pneumococcus adheres to the cells in the nose, invades these cells (so-called microinvasion), and activates them. Colonisation is a pre-requisite for disease, however, since disease is largely a dead end for S. pneumoniae, it remains unclear whether these processes are directly linked to disease progression. We have previously shown that if we introduce gene mutations into S. pneumoniae that affect key metabolic pathways, these bacteria retain their ability to colonize human and animal models without causing disease. We now show that these mutants retain their ability to microinvade epithelial cells in human and mouse models, and some may still cause inflammation, but are less able to pass through the epithelial barrier. However, although the attenuation of disease may be explained by the broad-ranging impact of these mutations on pneumococcal virulence, oxidative stress, and metabolism, they are not driven by a single determinant. Our findings suggest that pneumococcal microinvasion and immune activation are not necessarily pre-cursors to disease progression. This supports the idea that S. pneumoniae adapts and evolves to promote colonisation and ultimately transmission rather than cause disease.

Graphical AbstractS. pneumoniae colonisation is characterised by mucus association, epithelial adherence, microcolony formation and microinvasion - where the pneumococcus invades the epithelial barrier without causing disease. Although mutations in S. pneumoniae biosynthesis genes ({Delta}proABC and{Delta} fhs) attenuate disease in a murine model, they do not attenuate microinvasion in either experimental human pneumococcal challenge (EHPC), ex vivo or in vitro epithelial cells. Transmigration of the epithelial barrier is attenuated. These mutations show strain-dependent effects on both the epithelial and bacterial responses to infection. Factors such as epithelial cellular reorganisation, inflammation and caspase 8 activity alongside pneumococcal metabolic adaptation, virulence factor expression and response to stress are important components of these processes.



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=86 SRC="FIGDIR/small/545009v8_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Weight, C. M.</dc:creator>
<dc:creator>Pollara, G. M.</dc:creator>
<dc:creator>Betts, M.</dc:creator>
<dc:creator>Ragazzini, R.</dc:creator>
<dc:creator>Connor, M.</dc:creator>
<dc:creator>Ramos-Sevillano, E.</dc:creator>
<dc:creator>Reine, J.</dc:creator>
<dc:creator>Whelan, M.</dc:creator>
<dc:creator>Guerra-Assuncao, J. A.</dc:creator>
<dc:creator>Bonfanti, P.</dc:creator>
<dc:creator>Jolly, C.</dc:creator>
<dc:creator>Noursadeghi, M. C.</dc:creator>
<dc:creator>Ferreira, D.</dc:creator>
<dc:creator>Brown, J.</dc:creator>
<dc:creator>Heyderman, R. S.</dc:creator>
<dc:date>2023-06-15</dc:date>
<dc:identifier>doi:10.1101/2023.06.15.545009</dc:identifier>
<dc:title><![CDATA[Impact of Streptococcus pneumoniae biosynthesis gene mutations on epithelial microinvasion and cellular responses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.16.545259v1?rss=1">
<title>
<![CDATA[
The T766M-EGFR lung cancer mutation promotes tumor growth by exploiting newfound assembly mechanisms in ligand-free EGFR oligomers 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.16.545259v1?rss=1"
</link>
<description><![CDATA[
Epidermal growth factor receptor (EGFR) is central to cell growth in physiology and pathophysiologies, including non-small cell lung cancer (NSCLC). EGFR has been successfully targeted with tyrosine kinase inhibitor generations, but the missense secondary T766M mutation is a common cause of resistance. Overcoming this therapeutic challenge has been hindered by poor understanding of how T766M dysregulates EGFR function leading to tumor progression. Here we show that T766M amplifies tumor growth in vivo by exploiting newly discovered oligomer assembly mechanisms employed by wild type (WT)-EGFR to maintain ligand-independent basal phosphorylation. These mechanisms, also shared by drug-resistant exon 20 EGFR insertions, reveal tumor growth promoting functions for hitherto orphan transmembrane and kinase interfaces and for the ectodomain tethered conformation of EGFR. Placing our findings into the context of a ligand-free oligomer structure model, we provide a framework for future drug discovery directed at tackling EGFR mutations in cancer by disabling oligomer-assembling interactions.
]]></description>
<dc:creator>Martin-Fernandez, M. L.</dc:creator>
<dc:creator>Iyer, S.</dc:creator>
<dc:creator>Needham, S. R.</dc:creator>
<dc:creator>Galgadas, I.</dc:creator>
<dc:creator>Davis, B. M.</dc:creator>
<dc:creator>Roberts, S. K.</dc:creator>
<dc:creator>Man, R. C.</dc:creator>
<dc:creator>Zanetti-Domingues, L. C.</dc:creator>
<dc:creator>Clarke, D. T.</dc:creator>
<dc:creator>Fruhwirth, G. O.</dc:creator>
<dc:creator>Parker, P. J.</dc:creator>
<dc:creator>Rolfe, D. J.</dc:creator>
<dc:creator>Gervasio, F. L.</dc:creator>
<dc:date>2023-06-19</dc:date>
<dc:identifier>doi:10.1101/2023.06.16.545259</dc:identifier>
<dc:title><![CDATA[The T766M-EGFR lung cancer mutation promotes tumor growth by exploiting newfound assembly mechanisms in ligand-free EGFR oligomers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.20.545773v1?rss=1">
<title>
<![CDATA[
Structure-guided design and optimization of covalent VHL-targeted sulfonyl fluoride PROTACs 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.20.545773v1?rss=1"
</link>
<description><![CDATA[
O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=110 SRC="FIGDIR/small/545773v1_ufig1.gif" ALT="Figure 1">
View larger version (17K):
org.highwire.dtl.DTLVardef@da22e5org.highwire.dtl.DTLVardef@1290278org.highwire.dtl.DTLVardef@1611c0eorg.highwire.dtl.DTLVardef@7902d2_HPS_FORMAT_FIGEXP  M_FIG C_FIG Proteolysis-targeting chimeras (PROTACs) are heterobifunctional molecules that have emerged as a therapeutic modality to induce targeted protein degradation (TPD) by harnessing cellular proteolytic degradation machinery. PROTACs which ligand the E3 ligase in a covalent manner have attracted intense interest, however, covalent PROTACs with a broad protein of interest (POI) scope have proven challenging to discover by design. Here, we report structure-guided design and optimization of Von Hippel-Lindau (VHL) protein-targeted sulfonyl fluorides which covalently bind Ser110 in the HIF1 binding site. We demonstrate that their incorporation in bifunctional degraders induces targeted protein degradation of BRD4 or androgen receptor (AR) without further linker optimization. Our study discloses the first covalent VHL ligands which can be implemented directly in bifunctional degrader design expanding the substrate scope of covalent E3 ligase PROTACs.
]]></description>
<dc:creator>Shah, R. R.</dc:creator>
<dc:creator>De Vita, E.</dc:creator>
<dc:creator>Conole, D.</dc:creator>
<dc:creator>Sathyamurthi, P. S.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Fellows, E.</dc:creator>
<dc:creator>Fleites, C. M.</dc:creator>
<dc:creator>Queisser, M. A.</dc:creator>
<dc:creator>Harling, J. D.</dc:creator>
<dc:creator>Tate, E. W.</dc:creator>
<dc:date>2023-06-20</dc:date>
<dc:identifier>doi:10.1101/2023.06.20.545773</dc:identifier>
<dc:title><![CDATA[Structure-guided design and optimization of covalent VHL-targeted sulfonyl fluoride PROTACs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.19.544346v1?rss=1">
<title>
<![CDATA[
Wireless monitoring of respiration with EEG reveals relationships between respiration, behaviour and brain activity in freely moving mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.19.544346v1?rss=1"
</link>
<description><![CDATA[
Active sampling in the olfactory domain is an important aspect of mouse behaviour. Numerous methods are being used to measure active sampling behaviour, yet reliable observation of respiration in untethered, freely moving animals is challenging. So far, methods for measuring this have largely been restricted to head-fixed sniff monitoring, which makes it difficult to understand how sniff changes are related to natural mouse behaviour. Here, we implant a telemetry-based pressure sensor into the right jugular vein, which allows respiration to be measured via wireless thoracic pressure sensing in awake and freely moving, untethered mice. After verifying this technique against standard head-fixed respiration measurements, we investigated respiration patterns across a range of experiments in freely moving animals. Respiration frequency increased as mice voluntarily explored novel environmental cues. Combining wireless respiration measurements with EEG/EMG recording, we then used an evolving partial coherence analysis to uncover the direct relationships between respiration and brain activity in different frequency bands over the same exploration period. Finally, we examined respiration patterns across different vigilance states, revealing changes in passive respiration frequency across wakefulness, deep (NREM) sleep and dreaming (REM) sleep, and odour-triggered respiration increases in the absence of brain activity changes during NREM sleep. As it can be combined with behavioural assays and brain recordings, we anticipate that wireless respiration monitoring will be a valuable tool to increase our understanding of how mice use olfaction to process and interact with the environment around them.
]]></description>
<dc:creator>Dasgupta, D.</dc:creator>
<dc:creator>Schneider-Luftman, D.</dc:creator>
<dc:creator>Schaefer, A.</dc:creator>
<dc:creator>Harris, J. J.</dc:creator>
<dc:date>2023-06-21</dc:date>
<dc:identifier>doi:10.1101/2023.06.19.544346</dc:identifier>
<dc:title><![CDATA[Wireless monitoring of respiration with EEG reveals relationships between respiration, behaviour and brain activity in freely moving mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.17.545424v1?rss=1">
<title>
<![CDATA[
Dynamic X-chromosomal reactivation enhances female brain resilience 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.17.545424v1?rss=1"
</link>
<description><![CDATA[
While random X-chromosome inactivation in female cells of placental mammalians silences one allele of the majority of X-chromosomal genes, a considerable fraction is only incompletely and variably inactivated resulting in a tissue-specific pattern of mono- and biallelic expression. Here we used clonal human female induced pluripotent stem cells (iPSCs) allowing to trace the (in)activation status of the two X-chromosomes individually along neural differentiation trajectories. We discovered X-chromosome-wide locus- and lineage-specific dynamic usage of the two X-chromosomal alleles in female cells induced by differentiation. Leveraging iPSCs derived from patients with an X-linked neurodevelopmental disorder, we demonstrate that activation of alleles on the inactive X-chromosome can exert protective effects on the manifestation of disease phenotypes in female neural cells and tissue. Taken together, our data demonstrate that alleles on the inactive X-chromosome can serve as a critical reservoir reactivated during differentiation, thereby enhancing resilience of female neural tissue.
]]></description>
<dc:creator>Kaeseberg, S.</dc:creator>
<dc:creator>Bertin, M.</dc:creator>
<dc:creator>Menon, R.</dc:creator>
<dc:creator>Gabassi, E.</dc:creator>
<dc:creator>Todorov, H.</dc:creator>
<dc:creator>Frank, S.</dc:creator>
<dc:creator>Brennenstuhl, H.</dc:creator>
<dc:creator>Lohrer, B.</dc:creator>
<dc:creator>Winter, J.</dc:creator>
<dc:creator>Krummeich, J.</dc:creator>
<dc:creator>Winkler, J.</dc:creator>
<dc:creator>Winner, B.</dc:creator>
<dc:creator>Weis, E.</dc:creator>
<dc:creator>Hartwich, D.</dc:creator>
<dc:creator>Diederich, S.</dc:creator>
<dc:creator>Luck, K.</dc:creator>
<dc:creator>Gerber, S.</dc:creator>
<dc:creator>Lunt, P.</dc:creator>
<dc:creator>Berninger, B.</dc:creator>
<dc:creator>Falk, S.</dc:creator>
<dc:creator>Schweiger, S.</dc:creator>
<dc:creator>Karow, M.</dc:creator>
<dc:date>2023-06-21</dc:date>
<dc:identifier>doi:10.1101/2023.06.17.545424</dc:identifier>
<dc:title><![CDATA[Dynamic X-chromosomal reactivation enhances female brain resilience]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.20.545566v1?rss=1">
<title>
<![CDATA[
p97/VCP targets Toxoplasma gondii vacuoles for parasite restriction in interferon-stimulated human cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.20.545566v1?rss=1"
</link>
<description><![CDATA[
Infection with the parasite Toxoplasma gondii leads to production of interferon gamma (IFN{gamma}) that stimulates cells to upregulate defence proteins targeting the parasite for cell intrinsic elimination or growth restriction. Various host defence mechanisms operate at the parasitophorous vacuole (PV) in different human cell types leading to PV disruption, acidification, or membrane envelopment. Ubiquitin and p62 are players in all human host control mechanisms of Toxoplasma, but other unifying proteins have not been identified. Here, we show that p97/valosin-containing protein (VCP), as well as its associated proteins ANKRD13A and UBXD1 control Toxoplasma infection while recruited to the PV in IFN{gamma}-stimulated endothelial cells. Convergent deposition of ANKRD13A, p97/VCP and UBXD1 onto the same vacuole is dependent on vacuolar ubiquitination and observed within 2h post-infection. ANKRD13A, p97/VCP and UBXD1 all drive the acidification mechanism of the vacuole, which is the IFN{gamma}-dependent control pathway of Toxoplasma in endothelial cells. We assessed p97/VCP in Toxoplasma control in various human cells and demonstrate that p97/VCP is a universal IFN{gamma}-dependent host restriction factor targeting the Toxoplasma PV in epithelial (HeLa) and endothelial cells (HUVEC), fibroblasts (HFF) and macrophages (THP1).
]]></description>
<dc:creator>Clough, B.</dc:creator>
<dc:creator>Fisch, D.</dc:creator>
<dc:creator>Mize, T. H.</dc:creator>
<dc:creator>Encheva, V.</dc:creator>
<dc:creator>Snijders, A.</dc:creator>
<dc:creator>Frickel, E.</dc:creator>
<dc:date>2023-06-23</dc:date>
<dc:identifier>doi:10.1101/2023.06.20.545566</dc:identifier>
<dc:title><![CDATA[p97/VCP targets Toxoplasma gondii vacuoles for parasite restriction in interferon-stimulated human cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.25.545688v1?rss=1">
<title>
<![CDATA[
Comparative analysis of N-terminal cysteine dioxygenation and prolyl-hydroxylation as oxygen sensing pathways in mammalian cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.25.545688v1?rss=1"
</link>
<description><![CDATA[
In animals, adaptation to changes in cellular oxygen levels is coordinated largely by the 2-oxoglutarate dependent prolyl-hydroxylase domain (PHD) dioxygenase family, which regulate the stability of their hypoxia-inducible factor (HIF) substrates to promote expression of genes that adapt cells to hypoxia. Recently, 2-aminoethanethiol dioxygenase (ADO) was identified as a novel O2-sensing enzyme in animals. Through N-terminal cysteine dioxygenation and the N-degron pathway, ADO regulates the stability of a set of non-transcription factor substrates; the regulators of G-protein signalling 4, 5 and 16, and interleukin-32. Here, we set out to compare and contrast the in cellulo characteristics of ADO and PHD enzymes in an attempt to better understand their co-evolution in animals. We find that ADO operates to regulate the stability of its substrates rapidly and with similar O2-sensitivity to the PHD/HIF pathway. ADO appeared less sensitive to iron chelating agents or transition metal exposure than the PHD enzymes, possibly due to tighter catalytic-site Fe2+ coordination. Unlike the PHD/HIF pathway, the ADO/N-degron pathway was not subject to feedback by hypoxic induction of ADO and induction of ADO substrates was well sustained in response to prolonged hypoxia. The data also reveal strong interactions between proteolytic regulation of targets by ADO and transcriptional induction of those targets, that shape integrated cellular responses to hypoxia.
]]></description>
<dc:creator>Tian, Y.-M.</dc:creator>
<dc:creator>Holdship, P.</dc:creator>
<dc:creator>To, T. Q.</dc:creator>
<dc:creator>Ratcliffe, P. J.</dc:creator>
<dc:creator>Keeley, T. P.</dc:creator>
<dc:date>2023-06-26</dc:date>
<dc:identifier>doi:10.1101/2023.06.25.545688</dc:identifier>
<dc:title><![CDATA[Comparative analysis of N-terminal cysteine dioxygenation and prolyl-hydroxylation as oxygen sensing pathways in mammalian cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.25.546258v1?rss=1">
<title>
<![CDATA[
Clonal neural tube organoids self-organise floorplate through BMP-mediated cluster competition 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.25.546258v1?rss=1"
</link>
<description><![CDATA[
The neural tube (NT) has been a hallmark example of embryonic induction and patterning whereby the notochord induces an organiser, the floorplate, that secretes Sonic Hedgehog (SHH) to pattern the surrounding field of neural progenitors. On the other hand, NT organoids (NTOs) formed from embryonic stem cells (ESCs) undergo spontaneous floorplate formation and patterning in the absence of their normal embryonic inducers. Understanding how stem cells undergo regulative organiser formation is a central challenge in biology. Here, we investigated the self-organisation of a SHH-expressing floorplate organiser using clonal NTOs. Expression of FOXA2, a floorplate transcription factor, was initially spatially scattered before resolving into multiple clusters. These FOXA2+ clusters underwent competition and physical sorting, resulting in a stable "winning" floorplate. We identified BMP signalling as a key governor of long-range cluster competition. FOXA2+ clusters expressed BMP4 ligand suppressing FOXA2 in receiving cells, while simultaneously expressing the BMP-inhibitor NOGGIN to secure FOXA2+ cluster survival. Genetic mutation of Noggin perturbed the floorplate not only in NTOs but also in vivo at the mid-hindbrain region of the mouse NT. These results demonstrate how the floorplate can form autonomously without its well-known inducer, the notochord, suggesting redundant mechanisms ensuring robustness. Defining molecular pathways that govern organiser self-organisation is critical in harnessing the developmental plasticity of stem cells toward directed tissue engineering.
]]></description>
<dc:creator>Krammer, T.</dc:creator>
<dc:creator>Stuart, H. T.</dc:creator>
<dc:creator>Gromberg, E.</dc:creator>
<dc:creator>Ishihara, K.</dc:creator>
<dc:creator>Melchionda, M.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Costantini, E.</dc:creator>
<dc:creator>Lehr, S.</dc:creator>
<dc:creator>Cislo, D.</dc:creator>
<dc:creator>Arbanas, L.</dc:creator>
<dc:creator>Hoermann, A.</dc:creator>
<dc:creator>Neumueller, R. A.</dc:creator>
<dc:creator>Elvassore, N.</dc:creator>
<dc:creator>Siggia, E.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Kicheva, A.</dc:creator>
<dc:creator>Tanaka, E. M.</dc:creator>
<dc:date>2023-06-26</dc:date>
<dc:identifier>doi:10.1101/2023.06.25.546258</dc:identifier>
<dc:title><![CDATA[Clonal neural tube organoids self-organise floorplate through BMP-mediated cluster competition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.27.546642v1?rss=1">
<title>
<![CDATA[
Two-color coincidence single-molecule pull-down for the specific detection of disease-associated protein aggregates 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.27.546642v1?rss=1"
</link>
<description><![CDATA[
The misfolding and aggregation of protein is a characteristic of many neurodegenerative disorders, including Alzheimers and Parkinsons disease. The wide range of sizes and structures of oligomers and fibrils generated have previously been studied using single-molecule and super-resolution microscopy. These methods, however, tend to rely on the use of either directly labeled protein, or on the addition of non-specific amyloid stains, such as thioflavin-T. This has prevented the characterization of protein aggregate composition in complex biological samples. Here, we have developed a single-molecule two-color aggregate pull-down (STAPull) assay to overcome this challenge by probing immobilized proteins using orthogonally labeled antibodies targeting the same epitope. By looking at colocalized signals, we can eliminate monomeric protein, and specifically quantify aggregated proteins. Using the aggregation-prone alpha-synuclein protein as a model, we demonstrate that this approach can specifically detect aggregates with a limit of detection of 5 pM. Furthermore, we show that STAPull can be used in a range of samples, including in human biofluids. STAPull is generally applicable to protein aggregates from a variety of disorders, and will aid in the identification of biomarkers that are crucial in the effort to diagnose these diseases.
]]></description>
<dc:creator>Saleeb, R. S.</dc:creator>
<dc:creator>Leighton, C.</dc:creator>
<dc:creator>Lee, J.-E.</dc:creator>
<dc:creator>O'Shaughnessy, J.</dc:creator>
<dc:creator>Jeacock, K.</dc:creator>
<dc:creator>Chappard, A.</dc:creator>
<dc:creator>Cumberland, R.</dc:creator>
<dc:creator>Ball, S. R.</dc:creator>
<dc:creator>Sunde, M.</dc:creator>
<dc:creator>Clarke, D. J.</dc:creator>
<dc:creator>Piche, K.</dc:creator>
<dc:creator>McPhail, J. A.</dc:creator>
<dc:creator>Louwrier, A.</dc:creator>
<dc:creator>Angers, R.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Downey, P.</dc:creator>
<dc:creator>Kunath, T.</dc:creator>
<dc:creator>Horrocks, M. H.</dc:creator>
<dc:date>2023-06-29</dc:date>
<dc:identifier>doi:10.1101/2023.06.27.546642</dc:identifier>
<dc:title><![CDATA[Two-color coincidence single-molecule pull-down for the specific detection of disease-associated protein aggregates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.27.546656v1?rss=1">
<title>
<![CDATA[
Neuronal wiring diagram of an adult brain 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.27.546656v1?rss=1"
</link>
<description><![CDATA[
Connections between neurons can be mapped by acquiring and analyzing electron microscopic (EM) brain images. In recent years, this approach has been applied to chunks of brains to reconstruct local connectivity maps that are highly informative, yet inadequate for understanding brain function more globally. Here, we present the first neuronal wiring diagram of a whole adult brain, containing 5x107 chemical synapses between [~]130,000 neurons reconstructed from a female Drosophila melanogaster. The resource also incorporates annotations of cell classes and types, nerves, hemilineages, and predictions of neurotransmitter identities. Data products are available by download, programmatic access, and interactive browsing and made interoperable with other fly data resources. We show how to derive a projectome, a map of projections between regions, from the connectome. We demonstrate the tracing of synaptic pathways and the analysis of information flow from inputs (sensory and ascending neurons) to outputs (motor, endocrine, and descending neurons), across both hemispheres, and between the central brain and the optic lobes. Tracing from a subset of photoreceptors all the way to descending motor pathways illustrates how structure can uncover putative circuit mechanisms underlying sensorimotor behaviors. The technologies and open ecosystem of the FlyWire Consortium set the stage for future large-scale connectome projects in other species.
]]></description>
<dc:creator>Dorkenwald, S.</dc:creator>
<dc:creator>Matsliah, A.</dc:creator>
<dc:creator>Sterling, A. R.</dc:creator>
<dc:creator>Schlegel, P.</dc:creator>
<dc:creator>Yu, S.-c.</dc:creator>
<dc:creator>McKellar, C. E.</dc:creator>
<dc:creator>Lin, A.</dc:creator>
<dc:creator>Costa, M.</dc:creator>
<dc:creator>Eichler, K.</dc:creator>
<dc:creator>Yin, Y.</dc:creator>
<dc:creator>Silversmith, W.</dc:creator>
<dc:creator>Schneider-Mizell, C.</dc:creator>
<dc:creator>Jordan, C. S.</dc:creator>
<dc:creator>Brittain, D.</dc:creator>
<dc:creator>Halageri, A.</dc:creator>
<dc:creator>Kuehner, K.</dc:creator>
<dc:creator>Ogedengbe, O.</dc:creator>
<dc:creator>Morey, R.</dc:creator>
<dc:creator>Gager, J.</dc:creator>
<dc:creator>Kruk, K.</dc:creator>
<dc:creator>Perlman, E.</dc:creator>
<dc:creator>Yang, R.</dc:creator>
<dc:creator>Deutsch, D.</dc:creator>
<dc:creator>Bland, D.</dc:creator>
<dc:creator>Sorek, M.</dc:creator>
<dc:creator>Lu, R.</dc:creator>
<dc:creator>Macrina, T.</dc:creator>
<dc:creator>Lee, K.</dc:creator>
<dc:creator>Bae, J. A.</dc:creator>
<dc:creator>Mu, S.</dc:creator>
<dc:creator>Nehoran, B.</dc:creator>
<dc:creator>Mitchell, E.</dc:creator>
<dc:creator>Popovych, S.</dc:creator>
<dc:creator>Wu, J.</dc:creator>
<dc:creator>Jia, Z.</dc:creator>
<dc:creator>Castro, M.</dc:creator>
<dc:creator>Kemnitz, N.</dc:creator>
<dc:creator>Ih, D.</dc:creator>
<dc:creator>Bates, A. S.</dc:creator>
<dc:creator>Eckstein, N.</dc:creator>
<dc:creator>Funke, J.</dc:creator>
<dc:creator>Collman, F.</dc:creator>
<dc:creator>Bock, D. D.</dc:creator>
<dc:creator>Jefferis, G. S. X. E.</dc:creator>
<dc:creator>Seung, H. S.</dc:creator>
<dc:creator>Murthy, M.</dc:creator>
<dc:creator>the FlyWire Co</dc:creator>
<dc:date>2023-06-30</dc:date>
<dc:identifier>doi:10.1101/2023.06.27.546656</dc:identifier>
<dc:title><![CDATA[Neuronal wiring diagram of an adult brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.29.546675v1?rss=1">
<title>
<![CDATA[
Applying high-resolution spatial transcriptomics to characterise the amyloid plaque cell niche in Alzheimer's Disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.29.546675v1?rss=1"
</link>
<description><![CDATA[
The amyloid plaque cell niche is a pivotal hallmark of Alzheimers disease (AD). Where early spatial transcriptomics (ST) technologies have provided valuable information on transcriptomic alterations in the small tissue domains overlaying with amyloid plaques, they lacked cellular resolution. Here we compare two novel high-resolution ST platforms, CosMx and Stereo-seq, in their ability to characterize the cellular response in the amyloid plaque niche in an AD mouse model. Combining the results from both techniques empowered us to survey the highly variable microglial-astrocytic response across the amyloid plaque micro-environment and provided a first insight into how these responses could relate to neuronal transcriptomic alterations. This pilot study demonstrates the great potential of high-resolution ST, while simultaneously highlighting limitations that, when addressed, will unleash the full power of these techniques to map the progression of molecular and cellular changes in the brains of AD patients.
]]></description>
<dc:creator>Mallach, A.</dc:creator>
<dc:creator>Zielonka, M.</dc:creator>
<dc:creator>van Lieshout, V.</dc:creator>
<dc:creator>An, Y.</dc:creator>
<dc:creator>Khoo, J. H.</dc:creator>
<dc:creator>Vanheusden, M.</dc:creator>
<dc:creator>Chen, W.-T.</dc:creator>
<dc:creator>Moechars, D.</dc:creator>
<dc:creator>Arancibia-Carcamo, I. L.</dc:creator>
<dc:creator>Fiers, M.</dc:creator>
<dc:creator>De Strooper, B.</dc:creator>
<dc:date>2023-07-01</dc:date>
<dc:identifier>doi:10.1101/2023.06.29.546675</dc:identifier>
<dc:title><![CDATA[Applying high-resolution spatial transcriptomics to characterise the amyloid plaque cell niche in Alzheimer's Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.07.03.547533v1?rss=1">
<title>
<![CDATA[
The substrate quality of CK2 target sites has a determinant role on their function and evolution 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.07.03.547533v1?rss=1"
</link>
<description><![CDATA[
Most biological processes are regulated by peptide-recognition modules (PRMs) that bind to short linear motifs (SLiMs). Such interactions are rapidly reversible and often occur at low affinity. The protein kinase domain represents one such binding module, and known substrates may have full or only partial matches to the kinase recognition motif, a property known as  substrate quality. However, it is not yet clear whether differences in substrate quality represent neutral variation along the phosphosite sequence or if these differences have functional consequences that are subject to selection. We explore this question in detail for the acidophilic kinase CK2. CK2 is well-characterised, clinically important, and a fundamental enzyme for many aspects of cell biology. We show that optimal CK2 sites are phosphorylated at maximal stoichiometries and found in many conditions whereas minimal substrates are phosphorylated at lower stoichiometries, are more dynamic during the cell cycle, and have regulatory functions. Optimal CK2 sites also tend to be older and more conserved than minimal sites, and evolutionary simulations indicate that the substrate quality of CK2 phosphosites is often tuned by selection. For intermediate target sites, increases or decreases to substrate quality may be deleterious, which we demonstrate experimentally for a CK2 substrate at the kinetochore. The results together suggest that minimal and optimal phosphosites are strongly differentiated in terms of their functional and evolutionary properties.
]]></description>
<dc:creator>Bradley, D.</dc:creator>
<dc:creator>Garand, C.</dc:creator>
<dc:creator>Belda, H.</dc:creator>
<dc:creator>Gagnon-Arsenault, I.</dc:creator>
<dc:creator>Treeck, M.</dc:creator>
<dc:creator>Elowe, S.</dc:creator>
<dc:creator>Landry, C.</dc:creator>
<dc:date>2023-07-03</dc:date>
<dc:identifier>doi:10.1101/2023.07.03.547533</dc:identifier>
<dc:title><![CDATA[The substrate quality of CK2 target sites has a determinant role on their function and evolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.07.03.547468v1?rss=1">
<title>
<![CDATA[
Interrogation of RNA-protein interaction dynamics in bacterial growth 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.07.03.547468v1?rss=1"
</link>
<description><![CDATA[
Characterising RNA-protein interaction dynamics is fundamental to understand how bacteria respond to their environment. In this study, we have analysed the dynamics of 91% of the Escherichia coli expressed proteome and the RNA-interaction properties of 271 RNA-binding proteins (RBPs) at different growth phases. We find that 68% of RBPs differentially bind RNA across growth phases and characterise 17 previously unannotated proteins as bacterial RBPs including YfiF, a ncRNA-binding protein. While these new RBPs are mostly present in Proteobacteria, two of them have human orthologs in the form of mitochondrial proteins associated with rare metabolic disorders. Moreover, we reveal novel RBP functions for proteins such as the chaperone HtpG, a new stationary phase tRNA-binding protein. Altogether, we provide the first dynamic RBPome of a bacterium, showcasing how this approach can reveal the function of uncharacterised proteins, and identify critical RNA-protein interactions for cell growth which could inform new antimicrobial therapies.
]]></description>
<dc:creator>Monti, M.</dc:creator>
<dc:creator>Herman, R.</dc:creator>
<dc:creator>Davey, K.</dc:creator>
<dc:creator>Dawson, C. S.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Thomas, G. H.</dc:creator>
<dc:creator>Willis, A. E.</dc:creator>
<dc:creator>Lilley, K. S.</dc:creator>
<dc:creator>Villanueva, E.</dc:creator>
<dc:date>2023-07-03</dc:date>
<dc:identifier>doi:10.1101/2023.07.03.547468</dc:identifier>
<dc:title><![CDATA[Interrogation of RNA-protein interaction dynamics in bacterial growth]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.07.06.547989v1?rss=1">
<title>
<![CDATA[
Deep dynamical modelling of developmental trajectories with temporal transcriptomics 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.07.06.547989v1?rss=1"
</link>
<description><![CDATA[
Developmental cell fate decisions are dynamic processes driven by the complex behaviour of gene regulatory networks. A challenge in studying these processes using single-cell genomics is that the data provides only a static snapshot with no detail of dynamics. Metabolic labelling and splicing can provide time-resolved information, but current methods have limitations. Here, we present experimental and computational methods that overcome these limitations to allow dynamical modelling of gene expression from single-cell data. We developed sci-FATE2, an optimised metabolic labelling method that substantially increases data quality, and profiled approximately 45,000 embryonic stem cells differentiating into multiple neural tube identities. To recover dynamics, we developed velvet, a deep learning framework that extends beyond instantaneous velocity estimation by modelling gene expression dynamics through a neural stochastic differential equation system within a variational autoencoder. Velvet outperforms current velocity tools across quantitative benchmarks, and predicts trajectory distributions that accurately recapitulate underlying dataset distributions while conserving known biology. Velvet trajectory distributions capture dynamical aspects such as decision boundaries between alternative fates and correlative gene regulatory structure. Using velvet to provide a dynamical description of in vitro neural patterning, we highlight a process of sequential decision making and fate-specific patterns of developmental signalling. Together, these experimental and computational methods recast single-cell analyses from descriptions of observed data distributions to models of the dynamics that generated them, providing a new framework for investigating developmental gene regulation and cell fate decisions.
]]></description>
<dc:creator>Maizels, R. J.</dc:creator>
<dc:creator>Snell, D. M.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:date>2023-07-07</dc:date>
<dc:identifier>doi:10.1101/2023.07.06.547989</dc:identifier>
<dc:title><![CDATA[Deep dynamical modelling of developmental trajectories with temporal transcriptomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.07.13.548733v1?rss=1">
<title>
<![CDATA[
Cancer cell - fibroblast crosstalk signature promotes macrophage recruitment via HB-EGF and stromal EGFR/MEK signalling 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.07.13.548733v1?rss=1"
</link>
<description><![CDATA[
Interactions between cells in the tumor microenvironment (TME) shape cancer progression and patient outcomes. To gain new insights into how the TME influences cancer outcomes, we derive gene expression signatures indicative of signaling between stromal fibroblasts and cancer cells, and demonstrate their prognostic significance in multiple and independent squamous cell carcinoma cohorts. By leveraging information within the signatures, we discover that the HB-EGF/EGFR/MEK axis represents a hub of tumor - stroma crosstalk, promoting the expression of CSF2 and LIF and favoring the recruitment of macrophages. Together these analyses demonstrate the utility of our approach for interrogating the extent and consequences of TME crosstalk.
]]></description>
<dc:creator>Giangreco, G.</dc:creator>
<dc:creator>Rullan, A.</dc:creator>
<dc:creator>Naito, Y.</dc:creator>
<dc:creator>Biwas, D.</dc:creator>
<dc:creator>Liu, Y.-H.</dc:creator>
<dc:creator>Hooper, S.</dc:creator>
<dc:creator>Nenclares, P.</dc:creator>
<dc:creator>Bhide, S.</dc:creator>
<dc:creator>Cheang, M.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Hirata, E.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Melcher, A.</dc:creator>
<dc:creator>Harrington, K.</dc:creator>
<dc:creator>Sahai, E.</dc:creator>
<dc:date>2023-07-14</dc:date>
<dc:identifier>doi:10.1101/2023.07.13.548733</dc:identifier>
<dc:title><![CDATA[Cancer cell - fibroblast crosstalk signature promotes macrophage recruitment via HB-EGF and stromal EGFR/MEK signalling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.07.14.548982v1?rss=1">
<title>
<![CDATA[
Hif-2α programmes oxygen chemosensitivity in chromaffin cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.07.14.548982v1?rss=1"
</link>
<description><![CDATA[
The study of transcription factors that determine specialised neuronal functions has provided invaluable insights into the physiology of the nervous system. Peripheral chemoreceptors are neurone-like electro-physiologically excitable cells that link the oxygen content of arterial blood to the neuronal control of breathing. In the adult, this oxygen chemosensitivity is exemplified by the Type I cells of the carotid body and recent work has revealed one isoform of the transcription factor HIF, HIF-2, to have a non-redundant role in the development and function of that organ. Here we show that the activation of HIF-2, including isolated overexpression alone, is sufficient to induce oxygen chemosensitivity in the otherwise unresponsive adult adrenal medulla. This phenotypic change in the adrenal medulla was associated with retention of extra-adrenal paraganglioma-like tissues that resemble the foetal organ of Zuckerkandl and also manifest oxygen chemosensitivity. Acquisition of chemosensitivity was associated with changes in the adrenal medullary expression of classes of genes that are ordinarily characteristic of the carotid body, including G-protein regulators and atypical subunits of mitochondrial cytochrome oxidase. Overall, the findings suggest that, at least in certain tissues, HIF-2 acts as a phenotypic driver for cells that display oxygen chemosensitivity, providing a route to mechanistic understanding.
]]></description>
<dc:creator>Prange-Barczynska, M.</dc:creator>
<dc:creator>Jones, H.</dc:creator>
<dc:creator>Sugimoto, Y.</dc:creator>
<dc:creator>Cheng, X.</dc:creator>
<dc:creator>Lima, J.</dc:creator>
<dc:creator>Ratnayaka, I.</dc:creator>
<dc:creator>Douglas, G.</dc:creator>
<dc:creator>Buckler, K.</dc:creator>
<dc:creator>Ratcliffe, P.</dc:creator>
<dc:creator>Keeley, T.</dc:creator>
<dc:creator>Bishop, T.</dc:creator>
<dc:date>2023-07-14</dc:date>
<dc:identifier>doi:10.1101/2023.07.14.548982</dc:identifier>
<dc:title><![CDATA[Hif-2α programmes oxygen chemosensitivity in chromaffin cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.07.18.549490v1?rss=1">
<title>
<![CDATA[
Regulation of BCR-mediated Ca2+ mobilization by MIZ1-TIMBIM4 safeguards IgG1+ GC B cell positive selection 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.07.18.549490v1?rss=1"
</link>
<description><![CDATA[
The transition from IgM to affinity-matured IgG antibodies is vital for effective humoral immunity. This is facilitated by germinal centers (GCs) through affinity maturation and preferential accumulation of IgG+ B cells over IgM+ B cells. However, it is not known whether the positive selection of the different immunoglobulin isotypes within GCs varies in its dependency on specific transcriptional mechanisms. Here, we identified IgG1+ GC B cell transcription factor dependency using CRISPR-Cas9 and conditional mouse genetics. We found that MIZ1 was specifically required for IgG1+ GC B cell survival during positive selection, whereas IgM+ GC B cells were largely independent. Mechanistically, MIZ1 induced TMBIM4, an ancestral anti-apoptotic protein that regulated inositol trisphosphate receptor mediated Ca2+ mobilization downstream of IgG1. The MIZ1-TMBIM4 axis prevented mitochondrial dysfunction-induced IgG1+ GC cell death caused by excessive Ca2+ accumulation. This study uncovers a unique immunoglobulin isotype-specific dependency, on a hitherto unidentified mechanism in GC positive selection.
]]></description>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Toboso-Navasa, A.</dc:creator>
<dc:creator>Gunawan, A.</dc:creator>
<dc:creator>Camara, A.</dc:creator>
<dc:creator>Nakagawa, R.</dc:creator>
<dc:creator>Finsterbusch, K.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Newman, R.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Eilers, M.</dc:creator>
<dc:creator>Wack, A.</dc:creator>
<dc:creator>Tolar, P.</dc:creator>
<dc:creator>Toellner, K.-M.</dc:creator>
<dc:creator>Calado, D. P.</dc:creator>
<dc:date>2023-07-19</dc:date>
<dc:identifier>doi:10.1101/2023.07.18.549490</dc:identifier>
<dc:title><![CDATA[Regulation of BCR-mediated Ca2+ mobilization by MIZ1-TIMBIM4 safeguards IgG1+ GC B cell positive selection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.07.20.549875v1?rss=1">
<title>
<![CDATA[
Loss of REP1 impacts choroidal melanogenesis in choroideremia 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.07.20.549875v1?rss=1"
</link>
<description><![CDATA[
Choroideremia (CHM) is a rare X-linked chorioretinal dystrophy affecting the photoreceptors, retinal pigment epithelium (RPE) and choroid, however, the involvement of the choroid in disease progression is not fully understood. CHM is caused by mutations in the CHM gene, encoding the ubiquitously expressed Rab escort protein 1 (REP1). REP1 plays an important role in intracellular trafficking of vesicles, including melanosomes. In this study, we examined ultrastructure of the choroid in chmru848 fish and Chmnull/WT mouse models using transmission electron microscopy. Significant pigmentary disruptions were observed, with a lack of melanosomes in the choroid of chmru848fish from 4 days post fertilisation (4dpf). Total melanin and expression of melanogenesis genes tyr, tryp1a, mitf, dct and pmel were also reduced from 4dpf. In Chmnull/WT mice, choroidal melanosomes were significantly smaller at 1 month and at 1 year, eumelanin was reduced, and the choroid was thicker. The choroid in CHM patients was also examined using optical coherence tomography (OCT) and OCT- angiography (OCT-A) and the area of preserved choriocapillaris (CC) was found to be smaller than that of overlying photoreceptors, indicating that the choroid is degenerating at a faster rate. Histopathology of an enucleated eye from a 74-year-old CHM male patient revealed isolated areas of RPE but no associated underlying CC. Significant degenerative changes in the choroid of CHM patients and animal models are seen, highlighting the importance of administrative routes involving the choroid, such as suprachoroidal delivery. Pigmentary disruptions in CHM animal models reveal an important role for REP1 in melanogenesis, and drugs that improve melanin production represent a potential novel therapeutic avenue.
]]></description>
<dc:creator>Sarkar, H.</dc:creator>
<dc:creator>Tracey-White, D.</dc:creator>
<dc:creator>Hagag, A. M.</dc:creator>
<dc:creator>Burgoyne, T.</dc:creator>
<dc:creator>Jensen, L. D.</dc:creator>
<dc:creator>Edwards, M.</dc:creator>
<dc:creator>Moosajee, M.</dc:creator>
<dc:date>2023-07-20</dc:date>
<dc:identifier>doi:10.1101/2023.07.20.549875</dc:identifier>
<dc:title><![CDATA[Loss of REP1 impacts choroidal melanogenesis in choroideremia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.07.20.549584v1?rss=1">
<title>
<![CDATA[
A Taxonomically-informed Mass Spectrometry Search Tool for Microbial Metabolomics Data 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.07.20.549584v1?rss=1"
</link>
<description><![CDATA[
MicrobeMASST, a taxonomically-informed mass spectrometry (MS) search tool, tackles limited microbial metabolite annotation in untargeted metabolomics experiments. Leveraging a curated database of >60,000 microbial monocultures, users can search known and unknown MS/MS spectra and link them to their respective microbial producers via MS/MS fragmentation patterns. Identification of microbial-derived metabolites and relative producers, without a priori knowledge, will vastly enhance the understanding of microorganisms role in ecology and human health.
]]></description>
<dc:creator>Zuffa, S.</dc:creator>
<dc:creator>Schmid, R.</dc:creator>
<dc:creator>Bauermeister, A.</dc:creator>
<dc:creator>P. Gomes, P. W.</dc:creator>
<dc:creator>Caraballo-Rodriguez, A. M.</dc:creator>
<dc:creator>El Abiead, Y.</dc:creator>
<dc:creator>Aron, A. T.</dc:creator>
<dc:creator>Gentry, E. C.</dc:creator>
<dc:creator>Zemlin, J.</dc:creator>
<dc:creator>Meehan, M. J.</dc:creator>
<dc:creator>Avalon, N. E.</dc:creator>
<dc:creator>Cichewicz, R. H.</dc:creator>
<dc:creator>Buzun, E.</dc:creator>
<dc:creator>Carrillo Terrazas, M.</dc:creator>
<dc:creator>Hsu, C.-Y.</dc:creator>
<dc:creator>Oles, R.</dc:creator>
<dc:creator>Vasquez Ayala, A.</dc:creator>
<dc:creator>Zhao, J.</dc:creator>
<dc:creator>Chu, H.</dc:creator>
<dc:creator>Kuijpers, M. C. M.</dc:creator>
<dc:creator>Jackrel, S. L.</dc:creator>
<dc:creator>Tugizimana, F.</dc:creator>
<dc:creator>Nephali, L. P.</dc:creator>
<dc:creator>Dubery, I. A.</dc:creator>
<dc:creator>Madala, N. E.</dc:creator>
<dc:creator>Moreira, E. A.</dc:creator>
<dc:creator>Costa-Lotufo, L. V.</dc:creator>
<dc:creator>Lopes, N. P.</dc:creator>
<dc:creator>Rezende-Teixeira, P.</dc:creator>
<dc:creator>Jimenez, P. C.</dc:creator>
<dc:creator>Rimal, B.</dc:creator>
<dc:creator>Patterson, A. D.</dc:creator>
<dc:creator>Traxler, M. F.</dc:creator>
<dc:creator>Pessotti, R. d. C.</dc:creator>
<dc:creator>Alvarado-Villalobos, D.</dc:creator>
<dc:creator>Tamayo-Castillo, G.</dc:creator>
<dc:creator>Chaverri, P.</dc:creator>
<dc:creator>Escud</dc:creator>
<dc:date>2023-07-20</dc:date>
<dc:identifier>doi:10.1101/2023.07.20.549584</dc:identifier>
<dc:title><![CDATA[A Taxonomically-informed Mass Spectrometry Search Tool for Microbial Metabolomics Data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.07.26.550698v1?rss=1">
<title>
<![CDATA[
Investigating Brain Alterations in the Dp1Tyb Mouse Model of Down Syndrome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.07.26.550698v1?rss=1"
</link>
<description><![CDATA[
Down syndrome (DS) is one of the most common birth defects and the most prevalent genetic form of intellectual disability. DS arises from trisomy of chromosome 21, but its molecular and pathological consequences are not fully understood. In this study, we compared Dp1Tyb mice, a DS model, against their wild-type (WT) littermates of both sexes to investigate the impact of DS-related genetic abnormalities on the brain phenotype.

We performed in vivo whole brain magnetic resonance imaging (MRI) and hippocampal 1H magnetic resonance spectroscopy (MRS) on the animals at 3 months of age. Subsequently, ex vivo MRI scans and histological analyses were conducted post-mortem. Our findings unveiled distinct neuroanatomical and biochemical alterations in the Dp1Tyb brains.

Dp1Tyb brains exhibited a smaller surface area and a rounder shape compared to WT brains. Regional volumetric analysis revealed significant changes in 26 out of 72 examined brain regions, including the medial prefrontal cortex and dorsal hippocampus. These alterations were consistently observed in both in vivo and ex vivo imaging data. Additionally, high-resolution ex vivo imaging enabled us to investigate cerebellar layers and hippocampal subregions, revealing selective areas of decrease and remodelling in these structures.

An analysis of hippocampal metabolites revealed an elevation in glutamine and the glutamine/glutamate ratio in the Dp1Tyb mice compared to controls, suggesting a possible imbalance in the excitation/inhibition ratio. This was accompanied by the decreased levels of taurine. Histological analysis revealed fewer neurons in the hippocampal CA3 and DG layers, along with an increase in astrocytes and microglia. These findings recapitulate multiple neuroanatomical and biochemical features associated with DS, enriching our understanding of the potential connection between chromosome 21 trisomy and the resultant phenotype.
]]></description>
<dc:creator>Navacerrada, M. E. S.</dc:creator>
<dc:creator>Kim, E.</dc:creator>
<dc:creator>Siow, B.</dc:creator>
<dc:creator>Ma, D.</dc:creator>
<dc:creator>Gonzalez, L. R.</dc:creator>
<dc:creator>Simmons, C.</dc:creator>
<dc:creator>Hayward, D.</dc:creator>
<dc:creator>Gibbins, D.</dc:creator>
<dc:creator>Singh, N.</dc:creator>
<dc:creator>Strydom, A.</dc:creator>
<dc:creator>Fisher, E. M. C.</dc:creator>
<dc:creator>Tybulewicz, V. L. J.</dc:creator>
<dc:creator>Cash, D.</dc:creator>
<dc:date>2023-07-28</dc:date>
<dc:identifier>doi:10.1101/2023.07.26.550698</dc:identifier>
<dc:title><![CDATA[Investigating Brain Alterations in the Dp1Tyb Mouse Model of Down Syndrome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.07.26.550616v1?rss=1">
<title>
<![CDATA[
Sox3-null hypopituitarism depends on median eminence NG2-glia and is influenced by aspirin and gut microbiota 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.07.26.550616v1?rss=1"
</link>
<description><![CDATA[
The median eminence (ME), located at the base of the hypothalamus, is an essential centre of information exchange between the brain and the pituitary. We and others previously showed that mutations and duplications affecting the transcription factor SOX3/Sox3 result in hypopituitarism, and this is likely of hypothalamic origin. We demonstrate here that the absence of Sox3 predominantly affects the ME with phenotypes that first occur in juvenile animals, despite the embryonic onset of SOX3 expression. In the pituitary, reduction in hormone levels correlates with a lack of endocrine cell maturation. In parallel, ME NG2-glia renewal and oligodendrocytic differentiation potential are affected. We further show that low-dose aspirin treatment, which is known to affect NG2-glia, or changes in gut microbiota, rescue both proliferative defects and hypopituitarism in Sox3 mutants. Our study highlights a central role of NG2-glia for ME function during a transitional period of post-natal development, and indicates their sensitivity to environmental signals.
]]></description>
<dc:creator>Galichet, C.</dc:creator>
<dc:creator>Rizzoti, K.</dc:creator>
<dc:creator>Lovell-Badge, R.</dc:creator>
<dc:date>2023-07-28</dc:date>
<dc:identifier>doi:10.1101/2023.07.26.550616</dc:identifier>
<dc:title><![CDATA[Sox3-null hypopituitarism depends on median eminence NG2-glia and is influenced by aspirin and gut microbiota]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.07.29.551096v1?rss=1">
<title>
<![CDATA[
Proneural genes form a combinatorial code to diversify neocortical neural progenitor cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.07.29.551096v1?rss=1"
</link>
<description><![CDATA[
Neocortical neural progenitor cells (NPCs) are molecularly heterogeneous, yet the genes that confer distinct neuronal morphologies and connectivities during development are poorly understood. Here, we determined that a proneural gene combinatorial code diversifies cortical NPCs. By mining scRNA-seq data from murine embryonic and early postnatal cortices and generating trajectory inference models, we found that Neurog2 is predominant, and is transiently co-expressed with Ascl1 and/or Neurog1 during an apical-to-basal NPC transition state in NPCs with early pseudotime identities. To assess whether proneural gene pairs confer distinct properties, we first used Neurog2/Ascl1 reporter mice expressing unique reporters, revealing that NPCs have distinct cell division modes and cell cycle dynamics dependent on their proneural gene profile. To assess Neurog2/Neurog1 interactions, we used double knock-out mice and novel split-Cre transgenics crossed to a Rosa-diptheria-toxin-A line to delete double+ cells, showing Neurog1/Neurog2 are specifically required to generate early-born neurons and to maintain NPCs. Finally, in silico mutation of a cortical Neurog2-gene regulatory network and validation using Neurog1/Neurog2 mutant and  deleter mice, identified Bclllb and Nhlh2, expressed in early-born neurons, as dependent on Neurog1/Neurog2. Our data explains how proneural genes act combinatorically to diversify gene regulatory networks, thereby lineage restricting NPCs and creating cortical neuronal diversity.
]]></description>
<dc:creator>Moffat, A.</dc:creator>
<dc:creator>Oproescu, A.-M.</dc:creator>
<dc:creator>Okawa, S.</dc:creator>
<dc:creator>Han, S.</dc:creator>
<dc:creator>Vasan, L.</dc:creator>
<dc:creator>Ghazale, H.</dc:creator>
<dc:creator>Dennis, D. J.</dc:creator>
<dc:creator>Zinyk, D.</dc:creator>
<dc:creator>Guillemot, F.</dc:creator>
<dc:creator>del Sol, A.</dc:creator>
<dc:creator>Schuurmans, C.</dc:creator>
<dc:date>2023-07-31</dc:date>
<dc:identifier>doi:10.1101/2023.07.29.551096</dc:identifier>
<dc:title><![CDATA[Proneural genes form a combinatorial code to diversify neocortical neural progenitor cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.07.31.551250v1?rss=1">
<title>
<![CDATA[
MiR-155 epigenetically counteracts hypoxia for mitochondrial fitness during germinal center positive selection 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.07.31.551250v1?rss=1"
</link>
<description><![CDATA[
To increase antibody affinity against pathogens, positively selected GC-B cells initiate cell division in the light zone (LZ) of germinal centres (GCs). Among those, higher-affinity clones migrate to the dark zone (DZ) and vigorously proliferate by relying on oxidative phosphorylation (OXPHOS). However, it remains unknown how positively selected GC-B cells adapt their metabolism for cell division in the glycolysis-dominant, cell cycle arrest-inducing, hypoxic LZ microenvironment. Here, we show that microRNA (miR)-155 mediates metabolic reprogramming during positive selection to protect high-affinity clones. Transcriptome examination and mass spectrometry analysis revealed that miR-155 regulates H3K36me2 levels by directly repressing hypoxia-induced histone lysine demethylase, Kdm2a. This is indispensable for enhancing OXPHOS through optimizing the expression of vital nuclear mitochondrial genes under hypoxia. The miR-155-Kdm2a interaction is crucial to prevent excessive production of reactive oxygen species and apoptosis. Thus, miR-155-mediated epigenetic regulation promotes mitochondrial fitness in high-affinity clones, ensuring their expansion and consequently affinity maturation.
]]></description>
<dc:creator>Nakagawa, R.</dc:creator>
<dc:creator>Llorian-Sopena, M.</dc:creator>
<dc:creator>Varsani-brown, S.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Camarillo, J. M.</dc:creator>
<dc:creator>Barry, D.</dc:creator>
<dc:creator>George, R.</dc:creator>
<dc:creator>Blackledge, N. P.</dc:creator>
<dc:creator>Duddy, G.</dc:creator>
<dc:creator>Klose, R.</dc:creator>
<dc:creator>Turner, M.</dc:creator>
<dc:creator>Calado, D. P.</dc:creator>
<dc:date>2023-08-02</dc:date>
<dc:identifier>doi:10.1101/2023.07.31.551250</dc:identifier>
<dc:title><![CDATA[MiR-155 epigenetically counteracts hypoxia for mitochondrial fitness during germinal center positive selection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.02.551656v1?rss=1">
<title>
<![CDATA[
SCD inhibition eradicates AML displaying high de novo fatty acid desaturation and synergizes with chemotherapy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.02.551656v1?rss=1"
</link>
<description><![CDATA[
Identification of specific and therapeutically actionable vulnerabilities in acute myeloid leukaemia (AML) is needed to improve patients outcome. These features should be ideally present in many patients independently of mutational background. Here we identify de novo fatty acid (FA) desaturation, specifically stearoyl-CoA desaturase (SCD) inhibition, as a therapeutic vulnerability across multiple AML models in vitro and in vivo. We use the novel clinical grade SCD inhibitor SSI-4 to show that SCD inhibition induces AML cell death via pleiotropic effects, and sensitivity is based on their dependency on FA desaturation regardless of mutational profile. SSI-4 efficacy is enhanced by driving FA biosynthesis in vitro while stroma confers protective effects that extend to in vivo models. SCD inhibition increases DNA damage and its combination with standard DNA damage-inducing chemotherapy prolongs survival in aggressive murine AML models. Our work supports developing FA desaturase inhibitors in AML while stressing the importance of identifying predictive biomarkers of response and biologically validated combination therapies to realize their therapeutic potential.

One Sentence SummarySCD inhibition is toxic to AML cells with high rates of fatty acid desaturation and in combination with chemotherapy prolongs survival in murine AML models.
]]></description>
<dc:creator>Dembitz, V.</dc:creator>
<dc:creator>Lawson, H.</dc:creator>
<dc:creator>Burt, R.</dc:creator>
<dc:creator>Philippe, C.</dc:creator>
<dc:creator>James, S. C.</dc:creator>
<dc:creator>Atkinson, S.</dc:creator>
<dc:creator>Durko, J.</dc:creator>
<dc:creator>Wang, L. M.</dc:creator>
<dc:creator>Campos, J.</dc:creator>
<dc:creator>Magee, A. M.</dc:creator>
<dc:creator>Woodley, K.</dc:creator>
<dc:creator>Austin, M. J.</dc:creator>
<dc:creator>Rio-Machin, A.</dc:creator>
<dc:creator>Casado-Izquierdo, P.</dc:creator>
<dc:creator>Bewicke-Copley, F.</dc:creator>
<dc:creator>Rodriguez, G.</dc:creator>
<dc:creator>Pereira, D.</dc:creator>
<dc:creator>Oudejans, L.</dc:creator>
<dc:creator>Boet, E.</dc:creator>
<dc:creator>von Kriegsheim, A.</dc:creator>
<dc:creator>Schwaller, J.</dc:creator>
<dc:creator>Finch, A. J.</dc:creator>
<dc:creator>Pate, B.</dc:creator>
<dc:creator>Sarry, J.-E.</dc:creator>
<dc:creator>Tamburini, J.</dc:creator>
<dc:creator>Schuringa, J. J.</dc:creator>
<dc:creator>Hazlehurst, L.</dc:creator>
<dc:creator>Copland, J. A.</dc:creator>
<dc:creator>Yuneva, M.</dc:creator>
<dc:creator>Peck, B.</dc:creator>
<dc:creator>Cutillas, P.</dc:creator>
<dc:creator>Fitzgibbon, J.</dc:creator>
<dc:creator>Rouault-Pierre, K.</dc:creator>
<dc:creator>Kranc, K.</dc:creator>
<dc:creator>Gallipoli, P.</dc:creator>
<dc:date>2023-08-05</dc:date>
<dc:identifier>doi:10.1101/2023.08.02.551656</dc:identifier>
<dc:title><![CDATA[SCD inhibition eradicates AML displaying high de novo fatty acid desaturation and synergizes with chemotherapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.10.552581v1?rss=1">
<title>
<![CDATA[
Optimized dimerization of the PAR-2 RING domain drives cooperative and selective membrane recruitment for robust feedback-driven cell polarization 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.10.552581v1?rss=1"
</link>
<description><![CDATA[
The behavior of cell polarity networks is defined by the quantitative features of their constituent feedback circuits, which must be tuned to enable robust and stable polarization, while also ensuring that networks remain responsive to dynamically changing cellular states and/or spatial cues that arise during development. Using the PAR polarity network as a model, we demonstrate that these features are enabled by dimerisation of the polarity protein PAR-2 via ubiquitin-independent function of its N-terminal RING domain. Specifically, we combine theory and experiment to show that dimer affinity is optimized to achieve dynamic, selective, and cooperative recruitment of PAR-2 to the plasma membrane during polarization. Reducing dimerization results in loss of positive feedback and compromises robustness of symmetry-breaking, while enhanced dimerization renders the network less responsive due to kinetic trapping of PAR-2 on internal membranes and reduced sensitivity of PAR-2 to membrane displacement by the polarity kinase, aPKC/PKC-3. Thus, our data reveal how a dynamically oligomeric RING domain results in a cell polarity network that is both robust and responsive and highlight how tuning of oligomerization kinetics can serve as a general strategy for optimizing dynamic and cooperative intracellular targeting.
]]></description>
<dc:creator>Bland, T.</dc:creator>
<dc:creator>Hirani, N.</dc:creator>
<dc:creator>Briggs, D.</dc:creator>
<dc:creator>Rossetto, R.</dc:creator>
<dc:creator>Ng, K.</dc:creator>
<dc:creator>McDonald, N. Q.</dc:creator>
<dc:creator>Zwicker, D.</dc:creator>
<dc:creator>Goehring, N. W.</dc:creator>
<dc:date>2023-08-10</dc:date>
<dc:identifier>doi:10.1101/2023.08.10.552581</dc:identifier>
<dc:title><![CDATA[Optimized dimerization of the PAR-2 RING domain drives cooperative and selective membrane recruitment for robust feedback-driven cell polarization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.16.553490v1?rss=1">
<title>
<![CDATA[
Cortactin stabilizes actin branches by bridging activated Arp2/3 to its nucleated actin filament 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.16.553490v1?rss=1"
</link>
<description><![CDATA[
Regulation of the assembly and turnover of branched actin filament networks nucleated by the Arp2/3 complex is essential during many cellular processes including cell migration and membrane trafficking. Cortactin plays a key role in stabilizing actin filament branches by interacting with the Arp2/3 complex and actin filaments via its N-terminal Acidic domain (NtA) and 6.5 central unstructured 37 amino acid repeats, respectively 1, but the mechanism of this is unclear. We determined the structure of cortactin-stabilized Arp2/3 actin branches using cryo-electron microscopy. We find that cortactin interacts with the new daughter filament nucleated by the Arp2/3 complex at the branch site rather than the initial mother actin filament. Cortactin preferentially binds activated Arp3 in contrast to other nucleation promoting factors (NPFs) 2,3. Cortactin also stabilizes the F-actin-like interface of activated Arp3 with the first actin subunit of the new filament, and its central repeats extend along successive daughter filament subunits. Cortactin binding to Arp3 is incompatible with NPF interaction and its preference for activated Arp3 explains why it is retained at the actin branch. Our data have uncovered why cortactin displaces NPFs, while at the same time promoting synergy to regulate branched actin network dynamics.
]]></description>
<dc:creator>Liu, T.</dc:creator>
<dc:creator>Cao, L.</dc:creator>
<dc:creator>Mladenov, M.</dc:creator>
<dc:creator>JEGOU, A.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:creator>Moores, C.</dc:creator>
<dc:date>2023-08-16</dc:date>
<dc:identifier>doi:10.1101/2023.08.16.553490</dc:identifier>
<dc:title><![CDATA[Cortactin stabilizes actin branches by bridging activated Arp2/3 to its nucleated actin filament]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.14.553197v1?rss=1">
<title>
<![CDATA[
Therapeutic inhibition of monocyte recruitment prevents checkpoint inhibitor-induced hepatitis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.14.553197v1?rss=1"
</link>
<description><![CDATA[
Checkpoint inhibitor-induced hepatitis (CPI-hepatitis) is an emerging problem with the widening use of CPIs in cancer immunotherapy. Here, we developed a mouse model to characterise the mechanism of CPI-hepatitis and to therapeutically target key pathways driving this pathology. C57BL/6 wild-type (WT) mice were dosed with TLR9-agonist (TLR9-L) for hepatic priming combined with anti-CTLA-4 plus anti-PD-1 (CPI) or control (PBS) for up to 7 days. Co-administration of CPIs with TLR9-L induced liver pathology closely resembling human disease, with increased infiltration and clustering of granzyme B+perforin+CD8+ T cells and CCR2+ monocytes, 7 days post treatment. This was accompanied by apoptotic hepatocytes surrounding these clusters and elevated cytokeratin-18 and alanine transaminase plasma levels. Liver RNA sequencing identified key signalling pathways (JAK-STAT, NF-{kappa}B) and cytokine/chemokine networks (Ifn{gamma}, Cxcl9, Ccl2/Ccr2) as drivers of CPI-hepatitis. Using this model, we show that CD8+ T cells mediate hepatocyte damage in experimental CPI-hepatitis. However, their liver recruitment, clustering, and cytotoxic activity is dependent the presence of CCR2+ monocytes. Absence of hepatic monocyte recruitment in Ccr2rfp/rfp mice and CCR2 therapeutic inhibition by cenicriciroc (CVC) in WT mice prevented CPI-hepatitis. In conclusion, using this newly established mouse model, we demonstrate a central role of liver infiltrating CCR2+ monocyte interaction with cytotoxic CD8+ T cells in the pathogenesis of CPI-hepatitis and highlight novel therapeutic targets.
]]></description>
<dc:creator>Gudd, C. L. C.</dc:creator>
<dc:creator>Atkinson, S. R.</dc:creator>
<dc:creator>Mitchell, E.</dc:creator>
<dc:creator>Mawhin, M.-A.</dc:creator>
<dc:creator>Turajlic, S.</dc:creator>
<dc:creator>Larkin, J.</dc:creator>
<dc:creator>Thursz, M. R.</dc:creator>
<dc:creator>Goldin, R. D.</dc:creator>
<dc:creator>Powell, N.</dc:creator>
<dc:creator>Antoniades, C. G.</dc:creator>
<dc:creator>Woollard, K. J.</dc:creator>
<dc:creator>Possamai, L. A.</dc:creator>
<dc:creator>Triantafyllou, E.</dc:creator>
<dc:date>2023-08-16</dc:date>
<dc:identifier>doi:10.1101/2023.08.14.553197</dc:identifier>
<dc:title><![CDATA[Therapeutic inhibition of monocyte recruitment prevents checkpoint inhibitor-induced hepatitis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.16.553560v1?rss=1">
<title>
<![CDATA[
CDK activity at the centrosome regulates the cell cycle 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.16.553560v1?rss=1"
</link>
<description><![CDATA[
Cyclin-dependent kinases (CDKs) complexed with cyclins drive progression through the eukaryotic cell cycle. From yeast to human cells, cyclin-CDK localises to the centrosome, but the importance of this localisation is unclear. The conserved  hydrophobic patch substrate docking site on human Cyclin B1 and on the equivalent fission yeast Cdc13 mediates their localisation to the centrosome and the spindle-pole body (SPB, yeast centrosome equivalent). A hydrophobic patch mutant (HPM) of Cdc13 cannot enter mitosis, but whether this mitotic defect is due to defective SPB localisation or defective cyclin-substrate docking is unknown. Here we show that artificially restoring Cdc13HPM SPB localisation in fission yeast partially rescues both mitosis and defective CDK substrate phosphorylation at both the SPB and within the cytoplasm. In addition, we found that an HPM of the S-phase cyclin Cig2 has defective SPB localisation but is still able to perform bulk DNA synthesis. Our results demonstrate that the hydrophobic patch mediates the SPB localisation of both S- and M-phase cyclins, and that Cdc13 SPB localisation is essential for mitotic entry and for full phosphorylation of CDK substrates, supporting the view that the centrosome plays a role as a signalling hub regulating CDK cell cycle control.
]]></description>
<dc:creator>Roberts, E. L.</dc:creator>
<dc:creator>Kapadia, N.</dc:creator>
<dc:creator>Auchynnikava, T.</dc:creator>
<dc:creator>Basu, S.</dc:creator>
<dc:creator>Nurse, P.</dc:creator>
<dc:date>2023-08-16</dc:date>
<dc:identifier>doi:10.1101/2023.08.16.553560</dc:identifier>
<dc:title><![CDATA[CDK activity at the centrosome regulates the cell cycle]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.21.554177v1?rss=1">
<title>
<![CDATA[
Mutual homeostasis of charged proteins 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.21.554177v1?rss=1"
</link>
<description><![CDATA[
Protein dosage is regulated to maintain cellular homeostasis and health. The dosage of proteins containing disordered low complexity domains (LCDs) must be particularly well-controlled to prevent aberrant disease, yet no mechanism to maintain homeostasis has been identified1, 2. Here we report a mutual homeostatic mechanism that controls the concentration of such proteins, termed interstasis, in which proteins with similar LCDs co-regulate their combined dosage through collective negative feedback. We focused on the mechanism that exploits the fundamental multivalency of GA-rich RNA regions that encode charged LCDs, including those with arginine-enriched mixed charge domains (R-MCDs). Modest variations in the abundance of an R-MCD protein change the properties of nuclear speckles, a protein-RNA condensate, selectively trapping multivalent GA-rich mRNAs to promote their nuclear retention. This interstasis depends on conserved codon biases, shared by amniotes, which enhance the multivalency of GA-rich regions encoding charged LCDs. The threshold of interstasis is modulated by CLK kinases, which affect the nuclear speckle localisation of proteins such as TRA2B, key binder of GA-rich RNAs. Notably, many classes of LCDs are encoded by RNA regions containing multivalency-enhancing codon biases, each preferentially bound by specific proteins, suggesting that interstasis might co-regulate many classes of functionally related LCD-containing proteins through dose-sensitivity of various types of protein-RNA condensates.
]]></description>
<dc:creator>Faraway, R.</dc:creator>
<dc:creator>Heaven, N. C.</dc:creator>
<dc:creator>Digby, H.</dc:creator>
<dc:creator>Wilkins, O. G.</dc:creator>
<dc:creator>Chakrabarti, A. M.</dc:creator>
<dc:creator>Iosub, I. A.</dc:creator>
<dc:creator>Knez, L.</dc:creator>
<dc:creator>Ameres, S. L.</dc:creator>
<dc:creator>Plaschka, C.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:date>2023-08-21</dc:date>
<dc:identifier>doi:10.1101/2023.08.21.554177</dc:identifier>
<dc:title><![CDATA[Mutual homeostasis of charged proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.15.484379v1?rss=1">
<title>
<![CDATA[
Hypoxia inducible factors regulate infectious SARS-CoV-2, epithelial damage and respiratory symptoms in a hamster COVID-19 model. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.15.484379v1?rss=1"
</link>
<description><![CDATA[
Understanding the host pathways that define susceptibility to SARS-CoV-2 infection and disease are essential for the design of new therapies. Oxygen levels in the microenvironment define the transcriptional landscape, however the influence of hypoxia on virus replication and disease in animal models is not well understood. In this study, we identify a role for the hypoxic inducible factor (HIF) signalling axis to inhibit SARS-CoV-2 infection, epithelial damage and respiratory symptoms in Syrian hamsters. Pharmacological activation of HIF with the prolyl-hydroxylase inhibitor FG-4592 significantly reduced the levels of infectious virus in the upper and lower respiratory tract. Nasal and lung epithelia showed a reduction in SARS-CoV-2 RNA and nucleocapsid expression in treated animals. Transcriptomic and pathological analysis showed reduced epithelial damage and increased expression of ciliated cells. Our study provides new insights on the intrinsic antiviral properties of the HIF signalling pathway in SARS-CoV-2 replication that may be applicable to other respiratory pathogens and identifies new therapeutic opportunities.
]]></description>
<dc:creator>Wing, P. A.</dc:creator>
<dc:creator>Prange-Barczynska, M.</dc:creator>
<dc:creator>Cross, A.</dc:creator>
<dc:creator>Crotta, S.</dc:creator>
<dc:creator>Orbegozo Rubio, C.</dc:creator>
<dc:creator>Cheng, X.</dc:creator>
<dc:creator>Harris, J. M.</dc:creator>
<dc:creator>Zhuang, X.</dc:creator>
<dc:creator>Johnson, R. L.</dc:creator>
<dc:creator>Ryan, K. A.</dc:creator>
<dc:creator>Hall, Y.</dc:creator>
<dc:creator>Carroll, M. W.</dc:creator>
<dc:creator>Issa, F.</dc:creator>
<dc:creator>Balfe, P.</dc:creator>
<dc:creator>Wack, A.</dc:creator>
<dc:creator>Bishop, T.</dc:creator>
<dc:creator>Salguero, F. J.</dc:creator>
<dc:creator>McKeating, J. A.</dc:creator>
<dc:date>2022-03-15</dc:date>
<dc:identifier>doi:10.1101/2022.03.15.484379</dc:identifier>
<dc:title><![CDATA[Hypoxia inducible factors regulate infectious SARS-CoV-2, epithelial damage and respiratory symptoms in a hamster COVID-19 model.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.26.554941v1?rss=1">
<title>
<![CDATA[
Defective cytoskeletal dynamics underlies the essential role of MRTF-SRF in IL-2 delivery to CD8+ T cells during infectious challenge 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.26.554941v1?rss=1"
</link>
<description><![CDATA[
Paracrine IL-2 signalling underpins late primary CD8+ T cell expansion and differentiation that allow protection against viral infections, yet the requirements for effective delivery of IL-2 to recipient cells remain poorly understood. We show that the SRF transcription factor, a master regulator of cytoskeletal dynamics, is essential for the response to L. monocytogenes infection. SRF acts cell-autonomously with its actin-regulated MRTF cofactors Mrtfa and Mrtfb to sustain CD8+ effector T cell expansion and persistence of memory cells. MRTF-SRF activity is not required for initial TCR-mediated CD8+ T cell proliferation, but is necessary for subsequent IL-2 dependent expansion. Following TCR activation in vitro, Mrtfab-null CD8+ T cells produce IL-2 normally, but exhibit defective paracrine IL-2 signalling. Cluster formation by activated Mrtfab-null CD8+ T cells is impaired: clusters are smaller and less dense, have substantially reduced F-actin content, retain less IL-2, and exhibit defective cytoskeletal gene expression. Activated Mrtfab-null CD8+ T cells also exhibit defective homotypic clustering in vivo. The requirement for MRTF-SRF signalling for CD8+ T cell proliferation during infection thus reflects its involvement in cytoskeletal dynamics.
]]></description>
<dc:creator>Maurice, D.</dc:creator>
<dc:creator>Costello, P.</dc:creator>
<dc:creator>Gualdrini, F.</dc:creator>
<dc:creator>Frederico, B.</dc:creator>
<dc:creator>Treisman, R.</dc:creator>
<dc:date>2023-08-27</dc:date>
<dc:identifier>doi:10.1101/2023.08.26.554941</dc:identifier>
<dc:title><![CDATA[Defective cytoskeletal dynamics underlies the essential role of MRTF-SRF in IL-2 delivery to CD8+ T cells during infectious challenge]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.22.544179v1?rss=1">
<title>
<![CDATA[
Flow: a web platform and open database to analyse, store, curate and share bioinformatics data at scale 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.22.544179v1?rss=1"
</link>
<description><![CDATA[
Ever-increasing volumes of sequencing data offer potential for large-scale meta-analyses to address significant biological questions. However, challenges such as insufficient data processing information, data quality concerns, and issues related to accessibility and curation often present obstacles. Additionally, most experimental biologists lack the time and expertise needed to independently analyse, manage and share their own data. To overcome these hurdles, we present Flow, a web-based platform that links bioinformatic analysis and database solutions with a user-friendly interface and web API. Flow currently accommodates a range of genomics methods and further DSL2-compliant Nextflow pipelines can be added via a simple JSON schema file. Deployable on local systems or cloud services, an instance is freely accessible to academic researchers at https://flow.bio.
]]></description>
<dc:creator>Capitanchik, C.</dc:creator>
<dc:creator>Ireland, S.</dc:creator>
<dc:creator>Harston, A.</dc:creator>
<dc:creator>Cheshire, C.</dc:creator>
<dc:creator>Jones, D. M.</dc:creator>
<dc:creator>Lee, F. C. Y.</dc:creator>
<dc:creator>Ruiz de los Mozos, I.</dc:creator>
<dc:creator>Iosub, I. A.</dc:creator>
<dc:creator>Kuret, K.</dc:creator>
<dc:creator>Faraway, R.</dc:creator>
<dc:creator>Wilkins, O. G.</dc:creator>
<dc:creator>Arora, R.</dc:creator>
<dc:creator>Hallegger, M.</dc:creator>
<dc:creator>Modic, M.</dc:creator>
<dc:creator>Chakrabarti, A. M.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:date>2023-08-29</dc:date>
<dc:identifier>doi:10.1101/2023.08.22.544179</dc:identifier>
<dc:title><![CDATA[Flow: a web platform and open database to analyse, store, curate and share bioinformatics data at scale]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.29.555265v1?rss=1">
<title>
<![CDATA[
Unanticipated interacting features of human gut-associated lymphoid tissues link microbiota, intestinal immunity and autoimmunity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.29.555265v1?rss=1"
</link>
<description><![CDATA[
Gut-associated lymphoid tissue (GALT) is organised lymphoid tissue that is chronically activated by the intestinal microbiota. It generates the IgA response that is critical for intestinal homeostasis. By iterative application of multiplexed technologies, we identify enrichment of double-negative 2 (DN2:CD27-IgD-CD21loCD11chi) B cells in GALT, where they comprise the majority of intraepithelial and subepithelial B cells. We show that DN2 B cells in GALT interact with DC in the sub-epithelial dome that express DNASE1L3 and microbicides. Unlike in mice, DNASE1L3 in humans does not associate with apoptotic debris, but is located between sampled bacteria and host tissue where it is co-expressed with C1Q, consistent with management of bacterial debris. Thus we demonstrate that DN2 B cells that are otherwise associated with lupus nephritis, and DNASE1L3 and C1q that are lupus autoantigens, are microbiota-associated, interacting components of normal intestinal immunity.
]]></description>
<dc:creator>Montorsi, L.</dc:creator>
<dc:creator>Pitcher, M. J.</dc:creator>
<dc:creator>Zhao, Y.</dc:creator>
<dc:creator>Demonti, A.</dc:creator>
<dc:creator>Tull, T. J.</dc:creator>
<dc:creator>Dionisi, C.</dc:creator>
<dc:creator>Dhami, P.</dc:creator>
<dc:creator>Ellis, R. J.</dc:creator>
<dc:creator>Bishop, C.</dc:creator>
<dc:creator>Sanderson, J.</dc:creator>
<dc:creator>Jain, S.</dc:creator>
<dc:creator>D'Cruz, D.</dc:creator>
<dc:creator>Gibbons, D.</dc:creator>
<dc:creator>Winkler, T. H.</dc:creator>
<dc:creator>Bemark, M.</dc:creator>
<dc:creator>Ciccarelli, F. D.</dc:creator>
<dc:creator>Spencer, J.</dc:creator>
<dc:date>2023-08-31</dc:date>
<dc:identifier>doi:10.1101/2023.08.29.555265</dc:identifier>
<dc:title><![CDATA[Unanticipated interacting features of human gut-associated lymphoid tissues link microbiota, intestinal immunity and autoimmunity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.02.28.431778v1?rss=1">
<title>
<![CDATA[
DLL4 and PDGF-BB regulate migration of human iPSC-derived skeletal myogenic progenitors 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.02.28.431778v1?rss=1"
</link>
<description><![CDATA[
Muscle satellite stem cells (MuSCs) are responsible for skeletal muscle growth and regeneration. Despite their differentiation potential, human MuSCs have limited in vitro expansion and in vivo migration capacity, limiting their use in cell therapies for diseases affecting multiple skeletal muscle groups such as muscular dystrophies. Several protocols have been developed to derive progenitor cells similar to MuSCs from human induced pluripotent stem cells (hiPSCs), in order to establish a source of myogenic cells with controllable proliferation and differentiation capacity. However, currently available hiPSC myogenic derivatives also suffer from limitations of cell migration, ultimately delaying their clinical translation. Here we provide evidence that activation of NOTCH and PDGF pathways with DLL4 and PDGF-BB improves migration of hiPSC-derived myogenic progenitors in vitro. Transcriptomic and functional analyses demonstrate that this property is conserved across species and multiple hiPSC lines, including genetically-corrected hiPSC derivatives from a patient with Duchenne muscular dystrophy. DLL4 and PDGF-BB treatment had no negative impact on cell proliferation; cells maintained their myogenic memory, with differentiation fully rescued by NOTCH inhibition. RNAseq analysis indicate that pathways involved in cell migration are modulated in treated myogenic progenitors, consistent with results from functional profiling of cell motility at single cell resolution. Notably, treated cells also showed enhanced trans-endothelial migration in transwell assays. Enhancing extravasation is a key translational milestone for intravascular delivery of hiPSC myogenic derivatives: our study establishes the foundations of a transgene-free, developmentally inspired strategy to achieve this goal, moving hiPSCs one step closer to future muscle gene and cell therapies.
]]></description>
<dc:creator>Ferrari, G.</dc:creator>
<dc:creator>Choi, S.</dc:creator>
<dc:creator>Moyle, L. A.</dc:creator>
<dc:creator>Mackinlay, K.</dc:creator>
<dc:creator>Naouar, N.</dc:creator>
<dc:creator>Wells, C.</dc:creator>
<dc:creator>Muntoni, F.</dc:creator>
<dc:creator>Tedesco, F. S.</dc:creator>
<dc:date>2021-02-28</dc:date>
<dc:identifier>doi:10.1101/2021.02.28.431778</dc:identifier>
<dc:title><![CDATA[DLL4 and PDGF-BB regulate migration of human iPSC-derived skeletal myogenic progenitors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.09.06.556522v1?rss=1">
<title>
<![CDATA[
GOT1 primes the cellular response to hypoxia by supporting glycolysis and HIF1α stabilisation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.09.06.556522v1?rss=1"
</link>
<description><![CDATA[
Adaptation to chronic hypoxia occurs through changes in protein expression, which are controlled by hypoxia inducible factor 1a (HIF1) and are necessary for cancer cell survival. However, the mechanisms that enable cancer cells to adapt in early hypoxia, prior to full activation of HIF1, remain poorly understood. Here we show that aspartate transaminase 1 (GOT1), which supports NAD+ production by malate dehydrogenase 1 (MDH1), is required, in addition to reserve lactate dehydrogenase (LDH) capacity, for the HIF1-independent increase in glycolysis we observe early upon exposure of cells to hypoxia. Additionally, GOT1 maintains low -ketoglutarate levels, thereby limiting prolyl hydroxylase activity to promote HIF1 stabilisation in early hypoxia and robust HIF1 target gene expression in later hypoxia. Our findings reveal that, in normoxia, GOT1 maintains cells in a primed state and ready to support increased glycolysis and HIF1 stabilisation upon oxygen limitation, until other adaptive processes that require more time, are fully established.
]]></description>
<dc:creator>Anastasiou, D.</dc:creator>
<dc:creator>Grimm, F.</dc:creator>
<dc:creator>Asuaje, A.</dc:creator>
<dc:creator>Jain, A.</dc:creator>
<dc:creator>Silva dos Santos, M.</dc:creator>
<dc:creator>Kleinjung, J.</dc:creator>
<dc:creator>Nunes, P. M.</dc:creator>
<dc:creator>Gehrig, S.</dc:creator>
<dc:creator>Fets, L.</dc:creator>
<dc:creator>MacRae, J.</dc:creator>
<dc:date>2023-09-07</dc:date>
<dc:identifier>doi:10.1101/2023.09.06.556522</dc:identifier>
<dc:title><![CDATA[GOT1 primes the cellular response to hypoxia by supporting glycolysis and HIF1α stabilisation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.09.04.556195v1?rss=1">
<title>
<![CDATA[
Telomere-to-telomere Schizosaccharomyces japonicus genome assembly reveals hitherto unknown genome features 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.09.04.556195v1?rss=1"
</link>
<description><![CDATA[
Schizosaccharomyces japonicus belongs to the single-genus class Schizosaccharomycetes, otherwise known as  fission yeasts. As part of a composite model system with its widely studied S. pombe sister species, S. japonicus has provided critical insights into the workings and the evolution of cell biological mechanisms. Furthermore, its divergent biology makes S. japonicus a valuable model organism in its own right. However, the currently available short-read genome assembly contains gaps and has been unable to resolve centromeres and other repeat-rich chromosomal regions. Here we present a telomere-to-telomere long-read genome assembly of the S japonicus genome. This includes the three megabase-length chromosomes, with centromeres hundreds of kilobases long, rich in 5S ribosomal RNAs, transfer RNAs, long terminal repeats, and short repeats. We identify a gene-sparse region on chromosome 2 that resembles a 331 kb centromeric duplication. We revise the genome size of S. japonicus to at least 16.6 Mb and possibly up to 18.12 Mb, at least 30% larger than previous estimates. Our whole genome assembly will support the growing S. japonicus research community and facilitate research in new directions, including centromere and DNA repeat evolution, and yeast comparative genomics.

Take-awayO_LIA telomere-to-telomere genome assembly of the fission yeast S. japonicus
C_LIO_LIChromosome 2 harbours a previously unknown second centromere-like region
C_LIO_LIThe estimated genome size of S. japonicus may be up to 18.12 Mb
C_LI
]]></description>
<dc:creator>Etherington, G. J.</dc:creator>
<dc:creator>Nieduszynski, C.</dc:creator>
<dc:creator>Oliferenko, S.</dc:creator>
<dc:creator>Wu, P.-S.</dc:creator>
<dc:creator>Uhlmann, F.</dc:creator>
<dc:date>2023-09-06</dc:date>
<dc:identifier>doi:10.1101/2023.09.04.556195</dc:identifier>
<dc:title><![CDATA[Telomere-to-telomere Schizosaccharomyces japonicus genome assembly reveals hitherto unknown genome features]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.09.04.556188v1?rss=1">
<title>
<![CDATA[
Schizosaccharomyces versatilis represents a distinct evolutionary lineage. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.09.04.556188v1?rss=1"
</link>
<description><![CDATA[
The fission yeast species Schizosaccharomyces japonicus is currently divided into two varieties - S. japonicus var. japonicus and S. japonicus var. versatilis. Here we examine the var. versatilis isolate CBS5679. The CBS5679 genome shows 88% coding sequence identity to the reference genome of S. japonicus var. japonicus at the coding sequence level, with phylogenetic analyses suggesting that it has split from the S. japonicus lineage 25 million years ago. The CBS5679 genome contains a reciprocal translocation between chromosomes 1 and 2, together with several large inversions. The products of genes linked to the major translocation are associated with "metabolism" and "cellular assembly" ontology terms. We further show that CBS5679 does not generate viable progeny with the reference strain of S. japonicus. Although CBS5679 shares closer similarity to the "type" strain of var. versatilis as compared to S. japonicus, it is not identical to the type strain, suggesting population structure within var. versatilis. We recommend that the taxonomic status of S. japonicus var. versatilis is raised, with it being treated as a separate species, Schizosaccharomyces versatilis.

Take-awayO_LIThe taxonomic status of Schizosaccharomyces versatilis is addressed.
C_LIO_LIS. versatilis diverged from S. japonicus around 25 million years ago.
C_LIO_LIS. versatilis does not produce viable progeny in crosses with S. japonicus.
C_LIO_LIS. versatilis has a reciprocal translocation between chromosomes 1 and 2.
C_LIO_LIThe Gene Ontology terms for genes in the translocations are enriched for terms connected to "metabolism" and "cellular assembly".
C_LI
]]></description>
<dc:creator>Etherington, G. J.</dc:creator>
<dc:creator>Nieduszynski, C.</dc:creator>
<dc:creator>Oliferenko, S.</dc:creator>
<dc:creator>Gomez Gil, E.</dc:creator>
<dc:creator>Haerty, W.</dc:creator>
<dc:date>2023-09-06</dc:date>
<dc:identifier>doi:10.1101/2023.09.04.556188</dc:identifier>
<dc:title><![CDATA[Schizosaccharomyces versatilis represents a distinct evolutionary lineage.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.09.08.556779v1?rss=1">
<title>
<![CDATA[
Molecular model of a bacterial flagellar motor in situ reveals a "parts-list" of protein adaptations to increase torque 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.09.08.556779v1?rss=1"
</link>
<description><![CDATA[
One hurdle to understanding how molecular machines work, and how they evolve, is our inability to see their structures in situ. Here we describe a minicell system that enables in situ cryogenic electron microscopy imaging and single particle analysis to investigate the structure of an iconic molecular machine, the bacterial flagellar motor, which spins a helical propeller for propulsion. We determine the structure of the high-torque Campylobacter jejuni motor in situ, including the subnanometre-resolution structure of the periplasmic scaffold, an adaptation essential to high torque. Our structure enables identification of new proteins, and interpretation with molecular models highlights origins of new components, reveals modifications of the conserved motor core, and explain how these structures both template a wider ring of motor proteins, and buttress the motor during swimming reversals. We also acquire insights into universal principles of flagellar torque generation. This approach is broadly applicable to other membrane-residing bacterial molecular machines complexes.
]]></description>
<dc:creator>Drobnic, T.</dc:creator>
<dc:creator>Cohen, E. J.</dc:creator>
<dc:creator>Alzheimer, M.</dc:creator>
<dc:creator>Froschauer, K.</dc:creator>
<dc:creator>Svensson, S.</dc:creator>
<dc:creator>Singh, N.</dc:creator>
<dc:creator>Garg, S. G.</dc:creator>
<dc:creator>Henderson, L.</dc:creator>
<dc:creator>Umrekar, T.</dc:creator>
<dc:creator>Nans, A.</dc:creator>
<dc:creator>Ribardo, D.</dc:creator>
<dc:creator>Hochberg, G.</dc:creator>
<dc:creator>Hendrixson, D. R.</dc:creator>
<dc:creator>Sharma, C. M.</dc:creator>
<dc:creator>Rosenthal, P.</dc:creator>
<dc:creator>Beeby, M.</dc:creator>
<dc:date>2023-09-08</dc:date>
<dc:identifier>doi:10.1101/2023.09.08.556779</dc:identifier>
<dc:title><![CDATA[Molecular model of a bacterial flagellar motor in situ reveals a "parts-list" of protein adaptations to increase torque]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.09.12.557394v1?rss=1">
<title>
<![CDATA[
m6a methylation orchestrates IMP1 regulation of microtubules during human neuronal differentiation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.09.12.557394v1?rss=1"
</link>
<description><![CDATA[
Neuronal differentiation requires building a complex intracellular architecture, and therefore the coordinated regulation of defined sets of genes. RNA-binding proteins (RBPs) play a key role in this regulation. However, while their action on individual mRNAs has been explored in depth, the mechanisms used to coordinate expression of the gene programs shaping neuronal morphology are poorly understood. To address this, we analysed how the paradigmatic RBP IMP1 (IGF2BP1), an essential developmental factor, selects and regulates its RNA targets during the differentiation of human neurons. We performed a combination of system-wide and molecular analyses, revealing that IMP1 developmentally transitions to and directly regulates the expression of mRNAs encoding essential regulators of the microtubule network, a key component of neuronal morphology. Furthermore, we showed that m6A methylation drives the selection of specific IMP1 mRNA targets and their protein expression during the developmental transition from neural precursors to neurons, providing a molecular principle for the onset of target selectivity.
]]></description>
<dc:creator>Klein, P.</dc:creator>
<dc:creator>Harley, J.</dc:creator>
<dc:creator>Crook, H.</dc:creator>
<dc:creator>Serna, S. E.</dc:creator>
<dc:creator>Roumeliotis, T. I.</dc:creator>
<dc:creator>Choudhary, J. S.</dc:creator>
<dc:creator>Chakrabarti, A. M.</dc:creator>
<dc:creator>Luisier, R.</dc:creator>
<dc:creator>Patani, R.</dc:creator>
<dc:creator>Ramos, A.</dc:creator>
<dc:date>2023-09-12</dc:date>
<dc:identifier>doi:10.1101/2023.09.12.557394</dc:identifier>
<dc:title><![CDATA[m6a methylation orchestrates IMP1 regulation of microtubules during human neuronal differentiation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.09.15.556782v1?rss=1">
<title>
<![CDATA[
Sensory neuron population expansion enhances odour tracking through relaxed projection neuron adaptation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.09.15.556782v1?rss=1"
</link>
<description><![CDATA[
The evolutionary expansion of sensory neuron populations detecting important environmental cues is widespread, but functionally enigmatic. We investigated this phenomenon through comparison of homologous neural pathways of Drosophila melanogaster and its close relative Drosophila sechellia, an extreme specialist for Morinda citrifolia noni fruit. D. sechellia has evolved species-specific expansions in select, noni-detecting olfactory sensory neuron (OSN) populations, through multigenic changes. Activation and inhibition of defined proportions of neurons demonstrate that OSN population increases contribute to stronger, more persistent, noni-odor tracking behavior. These sensory neuron expansions result in increased synaptic connections with their projection neuron (PN) partners, which are conserved in number between species. Surprisingly, having more OSNs does not lead to greater odor-evoked PN sensitivity or reliability. Rather, pathways with increased sensory pooling exhibit reduced PN adaptation, likely through weakened lateral inhibition. Our work reveals an unexpected functional impact of sensory neuron expansions to explain ecologically-relevant, species-specific behavior.
]]></description>
<dc:creator>Takagi, S.</dc:creator>
<dc:creator>Abuin, L.</dc:creator>
<dc:creator>Stupski, S. D.</dc:creator>
<dc:creator>Arguello, J. R.</dc:creator>
<dc:creator>Prieto-Godino, L. L.</dc:creator>
<dc:creator>Stern, D. L.</dc:creator>
<dc:creator>Cruchet, S.</dc:creator>
<dc:creator>Alvarez-Ocana, R.</dc:creator>
<dc:creator>Wienecke, C. F. R.</dc:creator>
<dc:creator>van Breugel, F.</dc:creator>
<dc:creator>Auer, T. O.</dc:creator>
<dc:creator>Benton, R.</dc:creator>
<dc:date>2023-09-15</dc:date>
<dc:identifier>doi:10.1101/2023.09.15.556782</dc:identifier>
<dc:title><![CDATA[Sensory neuron population expansion enhances odour tracking through relaxed projection neuron adaptation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.09.14.557705v1?rss=1">
<title>
<![CDATA[
Blimp-1 and c-Maf regulate common and unique gene networks to protect against distinct pathways of pathobiont-induced colitis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.09.14.557705v1?rss=1"
</link>
<description><![CDATA[
Intestinal immune responses to commensals and pathogens are controlled by IL-10 to avoid intestinal immune pathology. We show that the transcription factors Blimp-1 (Prdm-1) and c-Maf are co-dominant regulators of Il10 in Foxp3+ regulatory T cells, but also negatively regulate proinflammatory cytokines in effector T cells. Mice with T cell-specific deletion of Prdm-1, Maf or the combination of both transcription factors did not develop inflammatory intestinal pathologies at the steady state. Double deficient Prdm1fl/flMaffl/flCd4Cre mice infected with Helicobacter hepaticus developed severe colitis with a major increase in TH1/NK/ILC1 effector genes in lamina propria leucocytes (LPLs), while Prdm1fl/flCd4Cre and Maffl/flCd4Cre mice showed mild/moderate pathology and a less-marked Type I effector response. LPLs from infected Maffl/flCd4Cre mice showed increased Il17a expression and an accompanying increase in granulocytes and myeloid cells, which was less marked in Prdm1fl/flMaffl/flCd4Cre mice, with increased T cell-myeloid-neutrophil interactions inferred from scRNA-seq analysis and confirmed by immunofluorescent analysis of colon sections. Genes over-expressed in human IBD showed differential expression in the LPL from infected mice in the absence of Prdm1 or Maf, revealing potential pathobiologic mechanisms of human disease.
]]></description>
<dc:creator>Alvarez-Martinez, M.</dc:creator>
<dc:creator>Cox, L. S.</dc:creator>
<dc:creator>Pearson, C. F.</dc:creator>
<dc:creator>Branchett, W. J.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Wu, X.</dc:creator>
<dc:creator>Slawinski, H.</dc:creator>
<dc:creator>Al Dibouni, A.</dc:creator>
<dc:creator>Samelis, V.</dc:creator>
<dc:creator>Gabrysova, L.</dc:creator>
<dc:creator>Priestnall, S. L.</dc:creator>
<dc:creator>Suarez-Bonnet, A.</dc:creator>
<dc:creator>Mikolajczak, A.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Powrie, F.</dc:creator>
<dc:creator>O'Garra, A.</dc:creator>
<dc:date>2023-09-16</dc:date>
<dc:identifier>doi:10.1101/2023.09.14.557705</dc:identifier>
<dc:title><![CDATA[Blimp-1 and c-Maf regulate common and unique gene networks to protect against distinct pathways of pathobiont-induced colitis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.09.18.558244v1?rss=1">
<title>
<![CDATA[
Congenital heart defects in Down syndrome are caused by increased dosage of DYRK1A 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.09.18.558244v1?rss=1"
</link>
<description><![CDATA[
Down syndrome (DS), trisomy 21, is a gene dosage disorder which results in multiple phenotypes including congenital heart defects (CHD). This clinically important pathology is caused by a third copy of one or more of the [~]230 genes on human chromosome 21 (Hsa21), but the identity of the causative dosage-sensitive genes is unknown and hence pathological mechanisms remain obscure. We show that embryonic hearts from human fetuses with DS and mouse models of DS have reduced expression of mitochondrial respiration and cell proliferation genes correlating with CHD. Using systematic genetic mapping, we determine that three copies of the Dyrk1a gene, encoding a serine/threonine protein kinase, are required to cause CHD. Reducing Dyrk1a copy number from three to two reverses defects in proliferation and mitochondrial respiration in embryonic cardiomyocytes and rescues septation defects in DS hearts. Furthermore, treatment of pregnant mice with a DYRK1A inhibitor developed for clinical use partially reduces the incidence of CHD among Dp1Tyb embryos. Thus, increased dosage of DYRK1A is required to impair mitochondrial function and cause CHD in DS, revealing a therapeutic target for this common human condition.

One Sentence SummaryIncreased dosage of DYRK1A causes mitochondrial dysfunction and congenital heart defects in Down syndrome and is ameliorated in utero by a drug.
]]></description>
<dc:creator>Lana-Elola, E.</dc:creator>
<dc:creator>Aoidi, R.</dc:creator>
<dc:creator>Llorian, M.</dc:creator>
<dc:creator>Gibbins, D.</dc:creator>
<dc:creator>Buechsenschuetz, C.</dc:creator>
<dc:creator>Bussi, C.</dc:creator>
<dc:creator>Flynn, H.</dc:creator>
<dc:creator>Gilmore, T.</dc:creator>
<dc:creator>Watson-Scales, S.</dc:creator>
<dc:creator>Haugsten Hansen, M.</dc:creator>
<dc:creator>Hayward, D.</dc:creator>
<dc:creator>Song, O.-R.</dc:creator>
<dc:creator>Brault, V.</dc:creator>
<dc:creator>Herault, Y.</dc:creator>
<dc:creator>Deau, E.</dc:creator>
<dc:creator>Meijer, L.</dc:creator>
<dc:creator>Snijders, A.</dc:creator>
<dc:creator>Gutierrez, M.</dc:creator>
<dc:creator>Fisher, E.</dc:creator>
<dc:creator>Tybulewicz, V.</dc:creator>
<dc:date>2023-09-19</dc:date>
<dc:identifier>doi:10.1101/2023.09.18.558244</dc:identifier>
<dc:title><![CDATA[Congenital heart defects in Down syndrome are caused by increased dosage of DYRK1A]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.09.21.558844v1?rss=1">
<title>
<![CDATA[
Multi-signal regulation of the GSK-3β homolog Rim11 governs meiosis entry in yeast 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.09.21.558844v1?rss=1"
</link>
<description><![CDATA[
Starvation of budding yeast diploid cells induces the cell-fate program that drives meiosis and spore formation. Transcription activation of early meiotic genes (EMGs) requires the transcription activator Ime1, its DNA-binding partner Ume6, and GSK-3{beta} kinase Rim11. Phosphorylation of Ume6 by Rim11 is key for EMG activation. We report that Rim11 integrates multiple input signals to control Ume6 phosphorylation and EMG transcription. Under nutrient-rich conditions PKA represses Rim11 to low levels while TORC1 keeps Rim11 localized to the cytoplasm. Inhibiting PKA and TORC1 induces Rim11 expression and nuclear localization. Remarkably, nuclear Rim11 is required, but not sufficient, for Rim11-dependent Ume6 phosphorylation. Additionally, Ime1 is an essential anchor protein for phosphorylating Ume6. Subsequently, Ume6-Ime1 coactivator complexes form that drive EMG transcription. Our results demonstrate how varied signalling inputs (PKA/TORC1/Ime1) integrated by Rim11 determine EMG expression and entry into meiosis. We propose that the signalling-regulatory network described here generates robustness in cell-fate control.
]]></description>
<dc:creator>Kociemba, J.</dc:creator>
<dc:creator>Joergensen, A. C. S.</dc:creator>
<dc:creator>Tadic, N.</dc:creator>
<dc:creator>Harris, A.</dc:creator>
<dc:creator>Sideri, T.</dc:creator>
<dc:creator>Chan, W. Y.</dc:creator>
<dc:creator>Ibrahim, F.</dc:creator>
<dc:creator>Unal, E.</dc:creator>
<dc:creator>Skehel, M.</dc:creator>
<dc:creator>Shahrezaei, V.</dc:creator>
<dc:creator>Arguello-Miranda, O.</dc:creator>
<dc:creator>van Werven, F. J.</dc:creator>
<dc:date>2023-09-22</dc:date>
<dc:identifier>doi:10.1101/2023.09.21.558844</dc:identifier>
<dc:title><![CDATA[Multi-signal regulation of the GSK-3β homolog Rim11 governs meiosis entry in yeast]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.09.22.558980v1?rss=1">
<title>
<![CDATA[
Regeneration of actin filament branches from the same Arp2/3 complex 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.09.22.558980v1?rss=1"
</link>
<description><![CDATA[
Branched actin filaments are found in many key cellular structures. Branches are nucleated by the Arp2/3 complex activated by nucleation-promoting factor (NPF) proteins and bound to the side of pre-existing  mother filaments. Over time, branches dissociate from their mother filament, leading to network reorganization and turnover, but this mechanism is less understood. Here, using microfluidics and purified proteins, we examined the dissociation of individual branches under controlled biochemical and mechanical conditions. We observe that Arp2/3 remains bound to the mother filament after most debranching events, even when accelerated by force. Unexpectedly, this mother-remaining Arp2/3 readily nucleates a new actin filament branch, without being activated anew by an NPF: it simply needs to exchange its nucleotide and bind an actin monomer. The protein GMF, which accelerates debranching, prevents branch re-nucleation. Our results suggest that actin filament re-nucleation can provide a self-repair mechanism, helping branched networks to sustain mechanical stress in cells over extended periods of time.
]]></description>
<dc:creator>Ghasemi, F.</dc:creator>
<dc:creator>Cao, L.</dc:creator>
<dc:creator>Mladenov, M.</dc:creator>
<dc:creator>Guichard, B.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:creator>Jegou, A.</dc:creator>
<dc:creator>Romet-Lemonne, G.</dc:creator>
<dc:date>2023-09-22</dc:date>
<dc:identifier>doi:10.1101/2023.09.22.558980</dc:identifier>
<dc:title><![CDATA[Regeneration of actin filament branches from the same Arp2/3 complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.09.25.559311v1?rss=1">
<title>
<![CDATA[
ARID3A coordinates the proliferation-differentiation switch of transit-amplifying cells in the intestine 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.09.25.559311v1?rss=1"
</link>
<description><![CDATA[
Intestinal stem cells (ISCs) at the crypt base divide and give rise to progenitor cells that have the capacity to proliferate and differentiate into various mature epithelial cell types in the transit-amplifying (TA) zone. Here, we identified the transcription factor ARID3A as a novel regulator of intestinal epithelial cell proliferation and differentiation at the TA compartment. We show that ARID3A forms an expression gradient from villus tip to the early progenitors at the crypts mediated by TGF-{beta} and WNT signalling. Intestinal epithelial-specific deletion of Arid3a reduces proliferation of TA cells. Bulk and single cell transcriptomic analysis shows increased enterocyte differentiation and reduced secretory cells in the Arid3a cKO intestine. Interestingly, upper-villus gene signatures of both enterocytes and secretory cells are enriched in the mutant intestine. We find that the enhanced enterocyte differentiation in the Arid3a cKO intestine is caused by increased binding of HNF1 and HNF4. Finally, we show that loss of Arid3a impairs irradiation-induced regenerative process by altering the dynamics of proliferation and apoptosis. Our findings imply that ARID3A may play a gatekeeping role in the TA compartment to maintain the "just-right" proliferation-to-differentiation ratio for tissue homeostasis and plasticity.
]]></description>
<dc:creator>Angelis, N.</dc:creator>
<dc:creator>Baulies, A.</dc:creator>
<dc:creator>Kucharska, A.</dc:creator>
<dc:creator>Kelly, G.</dc:creator>
<dc:creator>Sopena, M.</dc:creator>
<dc:creator>Boeing, S.</dc:creator>
<dc:creator>Li, V. S. W.</dc:creator>
<dc:date>2023-09-25</dc:date>
<dc:identifier>doi:10.1101/2023.09.25.559311</dc:identifier>
<dc:title><![CDATA[ARID3A coordinates the proliferation-differentiation switch of transit-amplifying cells in the intestine]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.09.25.557123v1?rss=1">
<title>
<![CDATA[
Bmp9 regulates Notch signaling and the temporal dynamics of angiogenesis via Lunatic Fringe 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.09.25.557123v1?rss=1"
</link>
<description><![CDATA[
In briefThe mechanisms regulating the signaling pathways involved in angiogenesis are not fully known. Ristori et al. show that Lunatic Fringe (LFng) mediates the crosstalk between Bone Morphogenic Protein 9 (Bmp9) and Notch signaling, thereby regulating the endothelial cell behavior and temporal dynamics of their identity during sprouting angiogenesis.

HighlightsO_LIBmp9 upregulates the expression of LFng in endothelial cells.
C_LIO_LILFng regulates the temporal dynamics of tip/stalk selection and rearrangement.
C_LIO_LILFng indicated to play a role in hereditary hemorrhagic telangiectasia.
C_LIO_LIBmp9 and LFng mediate the endothelial cell-pericyte crosstalk.
C_LI

Bone Morphogenic Protein 9 (Bmp9), whose signaling through Activin receptor-like kinase 1 (Alk1) is involved in several diseases, has been shown to independently activate Notch target genes in an additive fashion with canonical Notch signaling. Here, by integrating predictive computational modeling validated with experiments, we uncover that Bmp9 upregulates Lunatic Fringe (LFng) in endothelial cells (ECs), and thereby also regulates Notch activity in an inter-dependent, multiplicative fashion. Specifically, the Bmp9-upregulated LFng enhances Notch receptor activity creating a much stronger effect when Dll4 ligands are also present. During sprouting, this LFng regulation alters vessel branching by modulating the timing of EC phenotype selection and rearrangement. Our results further indicate that LFng can play a role in Bmp9-related diseases and in pericyte-driven vessel stabilization, since we find LFng contributes to Jag1 upregulation in Bmp9-stimulated ECs; thus, Bmp9-upregulated LFng results in not only enhanced EC Dll4-Notch1 activation, but also Jag1-Notch3 activation in pericytes.
]]></description>
<dc:creator>Ristori, T.</dc:creator>
<dc:creator>Thuret, R.</dc:creator>
<dc:creator>Hooker, E.</dc:creator>
<dc:creator>Quicke, P.</dc:creator>
<dc:creator>Lanthier, K.</dc:creator>
<dc:creator>Ntumba, K.</dc:creator>
<dc:creator>Aspalter, I. M.</dc:creator>
<dc:creator>Uroz, M.</dc:creator>
<dc:creator>Herbert, S. P.</dc:creator>
<dc:creator>Chen, C. S.</dc:creator>
<dc:creator>Larrivee, B.</dc:creator>
<dc:creator>Bentley, K.</dc:creator>
<dc:date>2023-09-25</dc:date>
<dc:identifier>doi:10.1101/2023.09.25.557123</dc:identifier>
<dc:title><![CDATA[Bmp9 regulates Notch signaling and the temporal dynamics of angiogenesis via Lunatic Fringe]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.09.27.559807v1?rss=1">
<title>
<![CDATA[
LRRK2 kinase dependent and independent function on endolysosomal repair promotes macrophage cell death 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.09.27.559807v1?rss=1"
</link>
<description><![CDATA[
LRRK2 is commonly mutated in Parkinsons disease and has cell type-specific mechanisms of activation and function. In macrophages, LRRK2 is associated with lysosomes and is activated following lysosomal damage. However, effects of pathogenic LRRK2-G2019S in macrophages are unknown. Here, using primary mouse and human iPSC-derived macrophage (iPSDM) models of LRRK2-G2019S, we defined the substrates of LRRK2 after lysosomal damage. Using phosphoproteomics we found that LRRK2-G2019S and wild-type macrophages showed similar levels of Rab phosphorylation after lysosomal damage, with the exceptions of Rab12 and Rab35, which were increased and decreased, respectively, in LRRK2-G2019S. LRRK2-G2019S macrophages showed a LRRK2 kinase activity-independent deficit in lysosomal membrane repair which resulted in more cell death and increased apoptosis. Importantly, we recapitulated this phenotype in iPSDM from patients carrying the G2019S mutation, but not in isogenic control iPSDM. Altogether, we define here the signaling downstream of G2019S in macrophages and identify susceptibility to cell death after lysosomal damage as an important phenotype of this mutation.
]]></description>
<dc:creator>Morrison, R.</dc:creator>
<dc:creator>Mihaylov, S. R. G.</dc:creator>
<dc:creator>Luk, C. H.</dc:creator>
<dc:creator>Rodgers, A.</dc:creator>
<dc:creator>Athanasiadi, N.</dc:creator>
<dc:creator>Morris, H.</dc:creator>
<dc:creator>Pellegrino, E.</dc:creator>
<dc:creator>Ultanir, S. K.</dc:creator>
<dc:creator>Gutierrez, M. G.</dc:creator>
<dc:date>2023-09-27</dc:date>
<dc:identifier>doi:10.1101/2023.09.27.559807</dc:identifier>
<dc:title><![CDATA[LRRK2 kinase dependent and independent function on endolysosomal repair promotes macrophage cell death]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.10.03.560673v1?rss=1">
<title>
<![CDATA[
Divergent downstream biosynthetic pathways are supported by L-cysteine synthases of Mycobacterium tuberculosis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.10.03.560673v1?rss=1"
</link>
<description><![CDATA[
Mycobacterium tuberculosiss (Mtb) autarkic lifestyle within the host involves rewiring its transcriptional networks to combat host-induced stresses. With the help of RNA-seq performed under various stress conditions, we identified that genes belonging to Mtb sulfur metabolism pathways are significantly upregulated during oxidative stress. Using an integrated approach of microbial genetics, transcriptomics, metabolomics, animal experiments, chemical inhibition, and rescue studies, we investigated the biological role of non-canonical L-cysteine synthases, CysM and CysK2. While transcriptome signatures of Rv{Delta}cysM and Rv{Delta}cysK2 appear similar under regular growth conditions, we observed unique transcriptional signatures when subjected to oxidative stress. We followed pool size and labelling (34S) of key downstream metabolites, viz. mycothiol and ergothioneine, to monitor L-cysteine biosynthesis and utilization. This revealed the significant role of distinct L-cysteine biosynthetic routes on redox stress and homeostasis. CysM and CysK2 independently facilitate Mtb survival by alleviating host-induced redox stress, suggesting they are not fully redundant during infection. With the help of genetic mutants and chemical inhibitors, we show that CysM and CysK2 serve as unique, attractive targets for adjunct therapy to combat mycobacterial infection.
]]></description>
<dc:creator>Khan, M.</dc:creator>
<dc:creator>Hunt, D. M.</dc:creator>
<dc:creator>Singha, B.</dc:creator>
<dc:creator>Kapoor, Y.</dc:creator>
<dc:creator>Singh, N.</dc:creator>
<dc:creator>Prasad, D. V. S.</dc:creator>
<dc:creator>Dharmarajan, S.</dc:creator>
<dc:creator>Sowpati, D. T.</dc:creator>
<dc:creator>de Carvalho, L.</dc:creator>
<dc:creator>Nandicoori, V. K.</dc:creator>
<dc:date>2023-10-05</dc:date>
<dc:identifier>doi:10.1101/2023.10.03.560673</dc:identifier>
<dc:title><![CDATA[Divergent downstream biosynthetic pathways are supported by L-cysteine synthases of Mycobacterium tuberculosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.10.11.561937v1?rss=1">
<title>
<![CDATA[
Aminoglycoside heteroresistance in Enterobacter cloacae is driven by the cell envelope stress response. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.10.11.561937v1?rss=1"
</link>
<description><![CDATA[
Enterobacter cloacae is a Gram-negative nosocomial pathogen of the ESKAPE priority group with increasing multi-drug resistance via the acquisition of resistance plasmids. However, E. cloacae can also display phenotypic antimicrobial resistance, such as heteroresistance or persistence. Here we report that E. cloacae ATCC 13047 and six strains isolated from patients with blood infections display heteroresistance or persistence to aminoglycosides. E. cloacae heteroresistance is transient, accompanied with formation of  petite colonies and increased MIC against gentamicin and other aminoglycosides used in the clinic, but not other antibiotic classes. To explore the underlying mechanisms, we performed RNA sequencing of heteroresistant bacteria, which revealed global gene-expression changes and a signature of the CpxRA cell envelope stress response. Deletion of the cpxRA two-component system abrogated aminoglycoside heteroresistance and petite colony formation, pointing to its indispensable role in phenotypic resistance. The introduction of a constitutively active allele of cpxA led to high aminoglycoside MICs, consistent with cell envelope stress driving these behaviours in E. cloacae. Cell envelope stress can be caused by environmental cues, including heavy metals. Indeed, bacterial exposure to copper increased gentamicin MIC in the wild type, but not the {Delta}cpxRA mutant. Moreover, copper exposure also elevated the gentamicin MICs of bloodstream isolates, suggesting that CpxRA- and copper-dependent aminoglycoside resistance is broadly conserved in E. cloacae strains. Altogether, we establish that E. cloacae relies on transcriptional reprogramming via the envelope stress response pathway for transient resistance to a major class of frontline antibiotic.

ImportanceEnterobacter cloacae is a bacterium that belongs to the ESKAPE priority group and an increasing threat worldwide due its multidrug resistance. E. cloacae can also display phenotypic resistance to antibiotics, leading to treatment failure. We report that sensitive strains of E. cloacae, including six strains isolated from patients with bloodstream infections, show heteroresistance or persistence to aminoglycoside antibiotics. These are important frontline microbicidal drugs used against Gram-negative bacterial infections, therefore understanding how resistance develops in sensitive strains is important. We show that aminoglycoside resistance is driven by the activation of the cell envelope stress response and transcriptional reprogramming via the CpxRA two-component system. Further, heterologous activation of envelope stress via copper, typically a heavy metal with antimicrobial actions, also increased aminoglycoside MICs in all tested strains of E. cloacae. Our study suggests phenotypic aminoglycoside resistance in E. cloacae could be broadly conserved and cautions against the undesirable effects of copper.
]]></description>
<dc:creator>Choi, A. J.</dc:creator>
<dc:creator>Bennison, D. J.</dc:creator>
<dc:creator>Kulkarni, E.</dc:creator>
<dc:creator>Sun, H.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Bradshaw, J.</dc:creator>
<dc:creator>Yeap, H. W.</dc:creator>
<dc:creator>Mishra, V.</dc:creator>
<dc:creator>Crespo-Puig, A.</dc:creator>
<dc:creator>Davies, F.</dc:creator>
<dc:creator>Sriskandan, S.</dc:creator>
<dc:creator>Shenoy, A. R.</dc:creator>
<dc:date>2023-10-11</dc:date>
<dc:identifier>doi:10.1101/2023.10.11.561937</dc:identifier>
<dc:title><![CDATA[Aminoglycoside heteroresistance in Enterobacter cloacae is driven by the cell envelope stress response.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.10.12.560630v1?rss=1">
<title>
<![CDATA[
Pan-cancer evolution signatures link clonal expansion to dynamic changes in the tumour immune microenvironment 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.10.12.560630v1?rss=1"
</link>
<description><![CDATA[
Cancer is an evolutionary process characterised by profound intra-tumour heterogeneity. Intra-tumour heterogeneity can be quantified using in silico estimates of cancer cell fractions of tumour-specific somatic mutations. Here we demonstrate a data-driven approach that uses cancer cell fraction distributions to identify 4 robust pan-cancer evolutionary signatures from an analysis of 4,146 individual tumour samples (TCGA) representing 17 distinct cancer types. Evolutionary signatures defined a continuum of cancer cell fractions representing neutral evolution, clonal expansion and fixation. Correlation of evolutionary signatures with programs representing distinct mutational and biological processes demonstrated that individual tumours enriched for clonal expansions and fixations were associated with immune evasion and distinct changes in the tumour immune microenvironment. We observed a dynamic switch between adaptive and innate immune processes as tumours undergo clonal fixation and escape immune surveillance. We also identify mutational processes underpinning different modes of tumour evolution and demonstrate that switching between adaptive and innate immune cell populations is accompanied by the clonal expansion of driver genes that modulate tumour-stroma interactions1.
]]></description>
<dc:creator>Yang, X.</dc:creator>
<dc:creator>Liu, W.</dc:creator>
<dc:creator>Mackintyre, G.</dc:creator>
<dc:creator>Loo, P. V.</dc:creator>
<dc:creator>Markowetz, F.</dc:creator>
<dc:creator>Bailey, P.</dc:creator>
<dc:creator>Yuan, K.</dc:creator>
<dc:date>2023-10-13</dc:date>
<dc:identifier>doi:10.1101/2023.10.12.560630</dc:identifier>
<dc:title><![CDATA[Pan-cancer evolution signatures link clonal expansion to dynamic changes in the tumour immune microenvironment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.10.19.563173v1?rss=1">
<title>
<![CDATA[
Cross-species imputation and comparison of single-cell transcriptomic profiles 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.10.19.563173v1?rss=1"
</link>
<description><![CDATA[
Cross-species comparison and prediction of gene expression profiles are important to understand regulatory changes during evolution and to transfer knowledge learned from model organisms to humans. Single-cell RNA-seq (scRNA-seq) profiles enable us to capture gene expression profiles with respect to variations among individual cells; however, cross-species comparison of scRNA-seq profiles is challenging because of data sparsity, batch effects, and the lack of one-to-one cell matching across species. Moreover, single-cell profiles are challenging to obtain in certain biological contexts, limiting the scope of hypothesis generation. Here we developed Icebear, a neural network framework that decomposes single-cell measurements into factors representing cell identity, species, and batch factors. Icebear enables accurate prediction of single-cell gene expression profiles across species, thereby providing high-resolution cell type and disease profiles in under-characterized contexts. Icebear also facilitates direct cross-species comparison of single-cell expression profiles for conserved genes that are located on the X chromosome in eutherian mammals but on autosomes in chicken. This comparison, for the first time, revealed evolutionary and diverse adaptations of X-chromosome upregulation in mammals.
]]></description>
<dc:creator>Zhang, R.</dc:creator>
<dc:creator>Yang, M.</dc:creator>
<dc:creator>Schreiber, J.</dc:creator>
<dc:creator>ODay, D. R.</dc:creator>
<dc:creator>Turner, J.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:creator>Disteche, C. M.</dc:creator>
<dc:creator>Deng, X.</dc:creator>
<dc:creator>Noble, W. S.</dc:creator>
<dc:date>2023-10-20</dc:date>
<dc:identifier>doi:10.1101/2023.10.19.563173</dc:identifier>
<dc:title><![CDATA[Cross-species imputation and comparison of single-cell transcriptomic profiles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.10.20.563362v1?rss=1">
<title>
<![CDATA[
Genome-wide CRISPR/Cas9 screen identifies MAT2A as a critical host factor for BK Polyomavirus 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.10.20.563362v1?rss=1"
</link>
<description><![CDATA[
BK polyomavirus (BKPyV) is a ubiquitous human pathogen that causes major complications for renal transplant patients (polyomavirus-associated nephropathy) and hematopoietic stem cell transplant patients (haemorrhagic cystitis). There are currently no effective antivirals available against BKPyV. As polyomaviruses are small DNA viruses that express very few proteins and utilise host DNA polymerases for their replication, there is limited possibility of targeting viral proteins for therapeutic intervention. As such, there is increasing interest in targeting host pathways to inhibit these viruses. To identify host genes required for BKPyV infection we have conducted the first genome-wide CRISPR/Cas9 screen in BKPyV infected cells. This led to the identification of methionine adenosyltransferase 2A (MAT2A), which we have validated as a host dependency factor for BKPyV infection. MAT2A is a druggable host enzyme that synthesises S-adenosylmethionine, the principal co-factor and methyl donor within cells. We have found that a small molecule inhibitor of MAT2A is a potent antiviral drug inhibiting BKPyV replication, offering a new therapeutic option for treatment of BKPyV diseases.
]]></description>
<dc:creator>Caller, L.</dc:creator>
<dc:creator>Ho, S.</dc:creator>
<dc:creator>Brown, K.</dc:creator>
<dc:creator>Dupont, L.</dc:creator>
<dc:creator>Zaletel, T.</dc:creator>
<dc:creator>Beale, R.</dc:creator>
<dc:creator>Lehner, P. J.</dc:creator>
<dc:creator>Crump, C. M.</dc:creator>
<dc:date>2023-10-21</dc:date>
<dc:identifier>doi:10.1101/2023.10.20.563362</dc:identifier>
<dc:title><![CDATA[Genome-wide CRISPR/Cas9 screen identifies MAT2A as a critical host factor for BK Polyomavirus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.10.22.563273v1?rss=1">
<title>
<![CDATA[
The history of chromosomal instability in genome doubled tumors 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.10.22.563273v1?rss=1"
</link>
<description><![CDATA[
Tumors frequently display high chromosomal instability (CIN) and contain multiple copies of genomic regions. Here, we describe GRITIC, a generic method for timing genomic gains leading to complex copy number states, using single-sample bulk whole-genome sequencing data. By applying GRITIC to 5,656 tumors, we found that non-parsimonious evolution is frequent in the formation of complex copy number states in genome-duplicated tumors. We measured CIN before and after genome duplication in human tumors and found that late genome doubling was followed by an increase in the rate of copy number gain. Copy number gains often accumulate as punctuated bursts, commonly after genome duplication. We infer that genome duplications typically affect the selection landscape of copy number losses, while only minimally impacting copy number gains. In summary, GRITIC is a novel copy number gain timing framework that permits the analysis of copy number evolution in chromosomally unstable tumors.

Statement of significanceComplex genomic gains are associated with whole-genome duplications, which are frequent across tumors, span a large fraction of their genomes, and are linked to poorer outcomes. GRITIC infers when these gains occur during tumor development, which will help to identify the genetic events that drive tumor evolution.
]]></description>
<dc:creator>Baker, T. M.</dc:creator>
<dc:creator>Lai, S.</dc:creator>
<dc:creator>Lesluyes, T.</dc:creator>
<dc:creator>Yan, H.</dc:creator>
<dc:creator>Verfaillie, A.</dc:creator>
<dc:creator>Dentro, S.</dc:creator>
<dc:creator>Lynch, A. R.</dc:creator>
<dc:creator>Bowes, A. L.</dc:creator>
<dc:creator>Pillay, N.</dc:creator>
<dc:creator>Flanagan, A. M.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Tarabichi, M.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:date>2023-10-25</dc:date>
<dc:identifier>doi:10.1101/2023.10.22.563273</dc:identifier>
<dc:title><![CDATA[The history of chromosomal instability in genome doubled tumors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.10.29.564563v1?rss=1">
<title>
<![CDATA[
Telomere protein arrays stall DNA loop extrusion by condensin 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.10.29.564563v1?rss=1"
</link>
<description><![CDATA[
DNA loop extrusion by SMC proteins is a key process underlying chromosomal organization. It is unknown how loop extruders interact with telomeres where chromosome ends are covered with a dense array of tens of neighboring DNA-binding proteins. Using complementary in vivo and in vitro single-molecule approaches, we study the interaction between loop-extruding condensin and arrays of Rap1, the double-stranded-DNA-binding telomeric protein of Saccharomyces cerevisiae. We show that dense linear Rap1 arrays can completely halt DNA loop extrusion, where the blocking efficiency depends on the array length and the DNA gap size between neighboring proteins. In cells, Rap1 arrays in the chromosome are found to act as contact insulators and to accumulate condensin at their borders, with direct implications for the resolution of dicentric chromosomes produced by telomere fusions. Our findings show that linear arrays of DNA-bound proteins can efficiently halt DNA loop extrusion by SMC proteins, which may impact a wide range of cellular processes from telomere functions to transcription and DNA repair.
]]></description>
<dc:creator>Analikwu, B. T.</dc:creator>
<dc:creator>Deshayes, A.</dc:creator>
<dc:creator>van der Torre, J.</dc:creator>
<dc:creator>Guerin, T.</dc:creator>
<dc:creator>Katan, A. J.</dc:creator>
<dc:creator>Beneut, C.</dc:creator>
<dc:creator>Barth, R.</dc:creator>
<dc:creator>Phipps, J.</dc:creator>
<dc:creator>Scolari, V.</dc:creator>
<dc:creator>Veaute, X.</dc:creator>
<dc:creator>Barrington, C.</dc:creator>
<dc:creator>Busso, D.</dc:creator>
<dc:creator>Uhlmann, F.</dc:creator>
<dc:creator>Dubrana, K.</dc:creator>
<dc:creator>Mattarocci, S.</dc:creator>
<dc:creator>Dekker, C.</dc:creator>
<dc:creator>Marcand, S.</dc:creator>
<dc:date>2023-10-29</dc:date>
<dc:identifier>doi:10.1101/2023.10.29.564563</dc:identifier>
<dc:title><![CDATA[Telomere protein arrays stall DNA loop extrusion by condensin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.10.27.563985v1?rss=1">
<title>
<![CDATA[
Atypical Protein Kinase C Promotes its own Asymmetric Localisation by Phosphorylating Cdc42 in Polarising Cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.10.27.563985v1?rss=1"
</link>
<description><![CDATA[
Atypical protein kinase C (aPKC) is a major regulator of cell polarity. Acting in conjunction with Par6, Par3 and the small GTPase Cdc42, aPKC becomes asymmetrically localised and drives the polarisation of cells. aPKC activity is crucial for its own asymmetric localisation, suggesting a hitherto unknown feedback mechanism contributing to polarisation. Here we show in the C. elegans zygote that the feedback relies on aPKC phosphorylation of Cdc42 at serine 71. The turnover of CDC-42 phosphorylation ensures optimal aPKC asymmetry and activity throughout polarisation by tuning Par6/aPKC association with Par3 and Cdc42. Moreover, turnover of Cdc42 phosphorylation regulates actomyosin cortex dynamics that are known to drive aPKC asymmetry. Given the widespread role of aPKC and Cdc42 in cell polarity, this form of self-regulation of aPKC may be vital for the robust control of polarisation in many cell types.

Key findings/graphical abstract- Phosphorylation of CDC-42 by aPKC accelerates aPKC dissociation from CDC-42, limiting aPKC activity
- CDC-42/aPKC dissociation promotes aPKC association with PAR-3 and, thereby, aPKC asymmetry due to actomyosin flow
- Cycling of CDC-42 phosphorylation fuels the exchange of aPKC between anteriorly transported PAR-3 and aPKC-active CDC-42 complexes
- Turnover of CDC-42 phosphorylation alternates its association with effectors, aPKC and MRCK-1, ensuring proper actomyosin dynamics




O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=136 SRC="FIGDIR/small/563985v2_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Packer, J.</dc:creator>
<dc:creator>Gubieda, A. G.</dc:creator>
<dc:creator>Brooks, A.</dc:creator>
<dc:creator>Deutz, L. N.</dc:creator>
<dc:creator>Squires, I.</dc:creator>
<dc:creator>Ellison, S.</dc:creator>
<dc:creator>Naganathan, S. R.</dc:creator>
<dc:creator>Wollman, A. J. M.</dc:creator>
<dc:creator>Dickinson, D. J.</dc:creator>
<dc:creator>Rodriguez, J.</dc:creator>
<dc:date>2023-10-28</dc:date>
<dc:identifier>doi:10.1101/2023.10.27.563985</dc:identifier>
<dc:title><![CDATA[Atypical Protein Kinase C Promotes its own Asymmetric Localisation by Phosphorylating Cdc42 in Polarising Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.11.06.565775v1?rss=1">
<title>
<![CDATA[
BDNF controls phosphorylation and transcriptional networks governing cytoskeleton organization and axonal regeneration 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.11.06.565775v1?rss=1"
</link>
<description><![CDATA[
The cell-intrinsic capacity of neurons to regenerate axons requires widespread coordination of the transcriptome, activation of multiple kinases, and reorganization of the cytoskeleton. Axonal repair is also influenced by extrinsic activating factors, such as neurotrophins. Here, we reveal that brain-derived neurotrophic factor (BDNF) amplifies multiple neuron-intrinsic programs to foster axonal regeneration in human motor neurons. Through metabolic RNA sequencing and phosphoproteomic profiling, we elucidate BDNF signalling and its role in axonal regeneration. We discover that BDNF controls RNA stability and transcriptional programs that converge with regeneration-associated gene (RAG) sets. We further unveil that BDNF governs the phosphorylation of multiple proteins essential for cytoskeletal dynamics, a major determinant of effective nerve regeneration. Using compartmentalized neuronal cultures, we demonstrate that the regeneration driven by BDNF depends on the axon-specific activation of ERK/RSK/S6K kinase pathway. We propose a model in which BDNF augments neuron-intrinsic pathways to drive axonal regeneration in human motor neurons.

TeaserBDNF aids nerve repair by fine-tuning the metabolism of RNA and by changing the building blocks of the nerve cell cytoskeleton.
]]></description>
<dc:creator>Vargas, J. N. S.</dc:creator>
<dc:creator>Brown, A.-L.</dc:creator>
<dc:creator>Sun, K.</dc:creator>
<dc:creator>Hagemann, C.</dc:creator>
<dc:creator>Geary, B.</dc:creator>
<dc:creator>Villarroel-Campos, D.</dc:creator>
<dc:creator>Bryce-Smith, S.</dc:creator>
<dc:creator>Zanovello, M.</dc:creator>
<dc:creator>Lombardo, M.</dc:creator>
<dc:creator>Ryandov, E.</dc:creator>
<dc:creator>Stepic, A.</dc:creator>
<dc:creator>Meysman, L.</dc:creator>
<dc:creator>Majewski, S.</dc:creator>
<dc:creator>Tosolini, A.</dc:creator>
<dc:creator>Secrier, M.</dc:creator>
<dc:creator>Keuss, M. J.</dc:creator>
<dc:creator>Serio, A.</dc:creator>
<dc:creator>Sleigh, J. N.</dc:creator>
<dc:creator>Fratta, P.</dc:creator>
<dc:creator>Schiavo, G.</dc:creator>
<dc:date>2023-11-06</dc:date>
<dc:identifier>doi:10.1101/2023.11.06.565775</dc:identifier>
<dc:title><![CDATA[BDNF controls phosphorylation and transcriptional networks governing cytoskeleton organization and axonal regeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.11.06.565813v1?rss=1">
<title>
<![CDATA[
Single cell RNA-seq reveals protracted germ line X chromosome reactivation dynamics directed by a PRC2 dependent mechanism 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.11.06.565813v1?rss=1"
</link>
<description><![CDATA[
Initiating soon after PGC specification, female germ cells undergo reactivation of the silenced X chromosome during genome wide reprogramming. However, the kinetics and dynamics of XCR in vivo have remained poorly understood. To address this here we perform a global appraisal of XCR using high-dimensional techniques. Using F1 B6 v CAST mouse embryos, we perform a detailed assessment, applying single-cell RNA-seq and chromatin profiling on germ cells purified from E10.5 to E16.5. While scRNA-seq profile showed that male and female germ cells are transcriptionally indistinct at E11.5, they are sexually dimorphic by E12.5, diverging further through development to E16.5. With allelic resolution, we show that the reactivating X chromosome is only partly active at E10.5, then reactivates gradually and reaches near parity in output to the constitutively active X chromosome at [~]E16.5 when developing oogonia are meiosis prophase I. Crucially, we show that sexually dimorphic dosage compensation patterns observed in germ cells, occur in tandem with an increase in the allelic proportion from the reactivating X chromosome. While Xist is extinguished from E10.5, the epigenetic memory of earlier XCI in female cells persists much longer, likely from self-sustained PRC2 complex (Ezh2 / Eed / Suz12) function. The reactivating X chromosome is enriched in the epigenetic silencing mark H3K27me3 at E13.5, which is removed by E16.5 permitting gene expression. Our findings link XCR, along with functional regulation of PRC2 in promoting female meiosis.
]]></description>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Lau, X.</dc:creator>
<dc:creator>Munusamy, P.</dc:creator>
<dc:creator>Sanchez, C. A. S.</dc:creator>
<dc:creator>Snell, D.</dc:creator>
<dc:creator>Sangrithi, M.</dc:creator>
<dc:date>2023-11-06</dc:date>
<dc:identifier>doi:10.1101/2023.11.06.565813</dc:identifier>
<dc:title><![CDATA[Single cell RNA-seq reveals protracted germ line X chromosome reactivation dynamics directed by a PRC2 dependent mechanism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.11.15.565967v1?rss=1">
<title>
<![CDATA[
Creation of de novo cryptic splicing for ALS/FTD precision medicine 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.11.15.565967v1?rss=1"
</link>
<description><![CDATA[
A system enabling the expression of therapeutic proteins specifically in diseased cells would be transformative, providing greatly increased safety and the possibility of pre-emptive treatment. Here we describe "TDP-REG", a precision medicine approach primarily for amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), which exploits the cryptic splicing events that occur in cells with TDP-43 loss-of-function (TDP-LOF) in order to drive expression specifically in diseased cells. In addition to modifying existing cryptic exons for this purpose, we develop a deep-learning-powered algorithm for generating customisable cryptic splicing events, which can be embedded within virtually any coding sequence. By placing part of a coding sequence within a novel cryptic exon, we tightly couple protein expression to TDP-LOF. Protein expression is activated by TDP-LOF in vitro and in vivo, including TDP-LOF induced by cytoplasmic TDP-43 aggregation. In addition to generating a variety of fluorescent and luminescent reporters, we use this system to perform TDP-LOF-dependent genomic prime editing to ablate the UNC13A cryptic donor splice site. Furthermore, we design a panel of tightly gated, autoregulating vectors encoding a TDP-43/Raver1 fusion protein, which rescue key pathological cryptic splicing events. In summary, we combine deep-learning and rational design to create sophisticated splicing sensors, resulting in a platform that provides far safer therapeutics for neurodegeneration, potentially even enabling preemptive treatment of at-risk individuals.

One-Sentence SummaryWe engineer TDP-43-regulated cryptic exons, enabling exceptionally precise activation of gene therapies in diseased neurons.
]]></description>
<dc:creator>Wilkins, O. G.</dc:creator>
<dc:creator>Chien, M. Z. Y. J.</dc:creator>
<dc:creator>Wlaschin, J. J.</dc:creator>
<dc:creator>Pisliakova, M.</dc:creator>
<dc:creator>Thompson, D.</dc:creator>
<dc:creator>Digby, H.</dc:creator>
<dc:creator>Simkin, R. L.</dc:creator>
<dc:creator>Diaz, J. A.</dc:creator>
<dc:creator>Mehta, P. R.</dc:creator>
<dc:creator>Keuss, M. J.</dc:creator>
<dc:creator>Zanovello, M.</dc:creator>
<dc:creator>Brown, A.-L.</dc:creator>
<dc:creator>Harley, P.</dc:creator>
<dc:creator>Darbey, A.</dc:creator>
<dc:creator>Karda, R.</dc:creator>
<dc:creator>Fisher, E. M. C.</dc:creator>
<dc:creator>Cunningham, T. J.</dc:creator>
<dc:creator>Le Pichon, C. E.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Fratta, P.</dc:creator>
<dc:date>2023-11-15</dc:date>
<dc:identifier>doi:10.1101/2023.11.15.565967</dc:identifier>
<dc:title><![CDATA[Creation of de novo cryptic splicing for ALS/FTD precision medicine]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.11.16.567403v1?rss=1">
<title>
<![CDATA[
3D-Imaging of synapses in neuronal tissues with synchrotron X-ray ptychography 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.11.16.567403v1?rss=1"
</link>
<description><![CDATA[
Maps of dense subcellular features in biological tissue are the key to understanding the structural basis of organ function. Electron microscopy provides the necessary resolution, yet - as electrons penetrate samples for only a few 100s of nm - requires physical sectioning or ablation, which strongly challenges anatomical investigations of entire organs such as mammalian brains. As demonstrated for the engineering and physical sciences, X-ray nanotomography represents a promising alternative for ultrastructural 3d imaging without physical sectioning1-15. Leveraging the high brilliance of 4th generation synchrotron X-ray sources, it has the potential to non-destructively image mm3-sized samples at ultrastructural resolution within a few days16. A fundamental barrier to application to the life sciences is that, when irradiated with high-intensity X-rays, biological samples deform and ultimately disintegrate, prohibiting reaching sufficient resolution. Here, we introduce a combination of engineering solutions which defeat this barrier for X-ray ptychography17, a coherent diffractive X-ray imaging technique. The solutions include a cryogenic sample stage with high stability, high-precision interferometric positioners and tailored non-rigid tomographic reconstruction algorithms18. Furthermore, adapting an epoxy resin developed for the nuclear and aerospace industry, we demonstrate radiation resistance to X-ray doses exceeding 1010 Gy. The resulting sub-40 nm isotropic resolution makes it possible to densely resolve axon bundles, boutons, dendrites and reliably identify synapses without physical sectioning. Moreover, we validated the X-ray technique using the current gold standard, namely focused ion beam scanning electron microscopy (FIB-SEM)19,20 to demonstrate intact ultrastructure in tissue volumes first imaged by X-rays. This unlocks the potential of X-ray tomography for high-resolution tissue imaging, coinciding with the transformative advancements of next-generation synchrotrons worldwide21.
]]></description>
<dc:creator>Bosch, C.</dc:creator>
<dc:creator>Diaz, A.</dc:creator>
<dc:creator>Holler, M.</dc:creator>
<dc:creator>Guizar-Sicairos, M.</dc:creator>
<dc:creator>Aidukas, T.</dc:creator>
<dc:creator>Pacureanu, A.</dc:creator>
<dc:creator>Mueller, E.</dc:creator>
<dc:creator>Peddie, C. J.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Menzel, A.</dc:creator>
<dc:creator>Wanner, A.</dc:creator>
<dc:creator>Schaefer, A.</dc:creator>
<dc:date>2023-11-17</dc:date>
<dc:identifier>doi:10.1101/2023.11.16.567403</dc:identifier>
<dc:title><![CDATA[3D-Imaging of synapses in neuronal tissues with synchrotron X-ray ptychography]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.11.21.568006v1?rss=1">
<title>
<![CDATA[
Nonlinear readout of spatial cues underlies robustness of asymmetric cell division 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.11.21.568006v1?rss=1"
</link>
<description><![CDATA[
A key challenge in the development of an organism is to maintain robust phenotypic outcomes in the face of perturbation. Yet, how such robust outcomes are encoded by developmental networks remains poorly explored. Here we use the C. elegans zygote as a model to understand sources of developmental robustness during PAR polarity-dependent asymmetric cell division. By quantitatively linking alterations in protein dosage to phenotype in individual embryos, we show that spatial information in the zygote is read out in a highly nonlinear fashion and, as a result, phenotypes are highly canalized against substantial variation in input signals. Specifically, our data point towards an intrinsic robustness of the conserved PAR polarity network that renders polarity axis specification resistant to variations in both the strength of upstream symmetry-breaking cues and PAR protein dosage. At the same time, we find that downstream pathways involved in cell size and fate asymmetry are similarly robust to dosage-dependent changes in the local concentrations of PAR proteins, implying non-trivial complexity in translating PAR signals into pathway outputs. We propose that "quantitative decoupling" of symmetry-breaking, polarity, and asymmetric division modules acts to suppress the accumulation of error as embryos move along this developmental trajectory, thereby ensuring that asymmetric division is robust to perturbation. Such modular organization of developmental networks is likely to be a general mechanism to achieve robust developmental outcomes.
]]></description>
<dc:creator>Rodrigues, N. T. L.</dc:creator>
<dc:creator>Bland, T.</dc:creator>
<dc:creator>Ng, K.</dc:creator>
<dc:creator>Hirani, N.</dc:creator>
<dc:creator>Goehring, N.</dc:creator>
<dc:date>2023-11-21</dc:date>
<dc:identifier>doi:10.1101/2023.11.21.568006</dc:identifier>
<dc:title><![CDATA[Nonlinear readout of spatial cues underlies robustness of asymmetric cell division]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.11.26.568719v1?rss=1">
<title>
<![CDATA[
CK-666 and CK-869 differentially inhibit Arp2/3 iso-complexes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.11.26.568719v1?rss=1"
</link>
<description><![CDATA[
The inhibitors, CK-666 and CK-869, are widely used to probe the function of actin nucleation by the Arp2/3 complex in vitro and in cells. However, in mammals, the Arp2/3 complex consists of 8 iso-complexes, as three of its subunits (Arp3, ArpC1, ArpC5) are encoded by two different genes. Here, we used recombinant Arp2/3 with defined composition to assess the activity of CK-666 and CK-869 against iso-complexes. We demonstrate that both inhibitors prevent linear actin filament formation when ArpC1A- or ArpC1B-containing complexes are activated by SPIN90. In contrast, inhibition of actin branching depends on iso-complex composition. Both drugs prevent actin branch formation by complexes containing ArpC1A, but only CK-869 can inhibit ArpC1B-containing complexes. Consistent with this, in bone marrow-derived macrophages which express low levels of ArpC1A, CK-869 but not CK-666, impacted phagocytosis and cell migration. CK-869 is also only able to inhibit Arp3-but not Arp3B-containing iso-complexes. Our findings have important implications for the interpretation of results using CK-666 and CK-869, given that the relative expression levels of ArpC1 and Arp3 isoforms in cells and tissues remains largely unknown.
]]></description>
<dc:creator>Cao, L.</dc:creator>
<dc:creator>Huang, S.</dc:creator>
<dc:creator>Basant, A.</dc:creator>
<dc:creator>Mladenov, M.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:date>2023-11-26</dc:date>
<dc:identifier>doi:10.1101/2023.11.26.568719</dc:identifier>
<dc:title><![CDATA[CK-666 and CK-869 differentially inhibit Arp2/3 iso-complexes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.11.30.569410v1?rss=1">
<title>
<![CDATA[
Cohesin chromatin loop formation by an extrinsic motor 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.11.30.569410v1?rss=1"
</link>
<description><![CDATA[
The ring-shaped cohesin complex topologically entraps two DNAs to establish sister chromatid cohesion1-3. Cohesin also shapes the interphase chromatin landscape with wide-ranging implications for gene regulation4-7, which cohesin is thought to achieve by actively extruding DNA loops without topologically entrapping DNA8-11. The  loop extrusion hypothesis finds motivation from in vitro observations12-14 - whether this process underlies in vivo chromatin loop formation remains untested. Here, using the budding yeast S. cerevisiae, we generate cohesin variants that have lost their ability to extrude DNA loops but retain their ability to topologically entrap DNA. Analysis of these variants suggests that in vivo chromatin loops form independently of loop extrusion. Instead, we find that transcription promotes loop formation, as well as acts as an extrinsic motor that expands these loops and defines their ultimate positions. Our results necessitate a re-evaluation of the loop extrusion model and point to an alternative mechanism for cohesin-dependent chromatin organisation. We propose that cohesin, akin to sister chromatid cohesion establishment at replication forks, forms chromatin loops by DNA-DNA capture at places of transcription, thus unifying cohesins two roles in chromosome segregation and interphase genome organisation.
]]></description>
<dc:creator>Guerin, T. M.</dc:creator>
<dc:creator>Barrington, C.</dc:creator>
<dc:creator>Pobegalov, G.</dc:creator>
<dc:creator>Molodtsov, M. I.</dc:creator>
<dc:creator>Uhlmann, F.</dc:creator>
<dc:date>2023-11-30</dc:date>
<dc:identifier>doi:10.1101/2023.11.30.569410</dc:identifier>
<dc:title><![CDATA[Cohesin chromatin loop formation by an extrinsic motor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.11.29.569216v1?rss=1">
<title>
<![CDATA[
Reconstructing oral cavity tumor evolution through brush biopsy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.11.29.569216v1?rss=1"
</link>
<description><![CDATA[
Oral premalignant lesions (OPLs) with genomic alterations have a heightened risk of evolving into oral squamous cell carcinoma (OSCC). Currently, genomic data are obtained through invasive tissue biopsy. Brush biopsy has been utilized for diagnosing dysplasia but its effectiveness in reflecting the complete genomic landscape of OPLs remains uncertain. This study investigates the potential of brush biopsy samples in accurately reconstructing the genomic profile of OPLs. We analyzed single nucleotide variants (SNVs), copy number aberrations (CNAs), and subclonal structures in paired tissue and brush biopsy samples from a patient with both OPL and OSCC lesions. The results showed that brush biopsy can effectively reflect about 90% of SNVs and similar CNA profiles as those found in tissue biopsies. It was specific, as normal oral epithelium didnt share these genomic alterations. Interestingly, brush biopsy revealed shared SNVs and CNAs between the distinct OPL and OSCC lesions, indicating a common ancestral origin. Subclonal reconstruction confirmed this shared ancestry, followed by divergent evolution of the lesions. These findings highlight the potential of brush biopsies in accurately representing the genomic profile of OPL and OSCC, proving useful in understanding tumor evolution.
]]></description>
<dc:creator>John, E.</dc:creator>
<dc:creator>Lesluyes, T.</dc:creator>
<dc:creator>Baker, T.</dc:creator>
<dc:creator>Tarabichi, M.</dc:creator>
<dc:creator>Gillenwater, A.</dc:creator>
<dc:creator>Wang, J. R.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>Zhao, X.</dc:creator>
<dc:date>2023-12-01</dc:date>
<dc:identifier>doi:10.1101/2023.11.29.569216</dc:identifier>
<dc:title><![CDATA[Reconstructing oral cavity tumor evolution through brush biopsy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.01.569635v1?rss=1">
<title>
<![CDATA[
Antigenic and genetic diversity of H1 and H3 influenza A viruses in swine in Brazil 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.01.569635v1?rss=1"
</link>
<description><![CDATA[
BackgroundInfluenza A virus (IAV) circulates within human and swine populations, and pigs are considered intermediate hosts for the generation of IAV with pandemic potential. Surveillance and characterization of IAVs circulating in pig populations are crucial to strain match vaccines to control IAV transmission in pigs and quantify pandemic potential to humans.

MethodsHere, we characterized the genetic and antigenic diversity of IAVs circulating in Brazilian swine between 2010-2018. Phylogenetic maximum-likelihood trees were generated for 84 Brazilian hemagglutinin (HA) gene segments. Hemagglutination inhibition (HI) assay data was used with antigenic cartography to quantify the antigenic differences among representative H1 and H3 swine viruses and relative cross-reactivity between these viruses and human seasonal vaccine strains.

ResultsWe identified two genetic lineages of H1 viruses derived from separate human-to-swine transmission events (H1 1B lineage, clades 1B.2.3 and 1B.2.4), an H3 lineage that has diversified into two genetic clades (H3 1990.5.1 and 1990.5.2), and HA genes associated with the 2009 H1N1 pandemic. There was limited cross-reactivity between circulating swine lineages and significant antigenic variation within lineage.

ConclusionsThe antigenic diversity among endemic IAV in swine indicates a need for regional strain-specific vaccination strategies in Brazil. Our data supports the need for systematic genomic surveillance and characterization in Brazil to improve the efficacy of swine vaccines and quantify the pandemic potential of endemic swine influenza A viruses.
]]></description>
<dc:creator>Lopes, S.</dc:creator>
<dc:creator>Anderson, T. K.</dc:creator>
<dc:creator>Schaefer, R.</dc:creator>
<dc:creator>Tochetto, C.</dc:creator>
<dc:creator>Gava, D.</dc:creator>
<dc:creator>Cantao, M. E.</dc:creator>
<dc:creator>Ciacci-Zanella, J. R.</dc:creator>
<dc:creator>Vincent Baker, A. L.</dc:creator>
<dc:creator>Lewis, N. S.</dc:creator>
<dc:date>2023-12-02</dc:date>
<dc:identifier>doi:10.1101/2023.12.01.569635</dc:identifier>
<dc:title><![CDATA[Antigenic and genetic diversity of H1 and H3 influenza A viruses in swine in Brazil]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.04.569933v1?rss=1">
<title>
<![CDATA[
Four-Core Genotypes mice harbour a 3.2MB X-Y translocation that perturbs Tlr7 dosage 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.04.569933v1?rss=1"
</link>
<description><![CDATA[
The Four Core Genotypes (FCG) is a mouse model system heavily used to disentangle the function of sex chromosomes and hormones. We report that a copy of a 3.2 MB region of the X chromosome has translocated to the YSry- chromosome and thus increased the expression of multiple genes including the auto-immune master regulator Tlr7. This previously-unreported X-Y translocation complicates the interpretation of studies reliant on FCG mice.
]]></description>
<dc:creator>Panten, J.</dc:creator>
<dc:creator>Del Prete, S.</dc:creator>
<dc:creator>Cleland, J. P.</dc:creator>
<dc:creator>Saunders, L. M.</dc:creator>
<dc:creator>van Riet, J.</dc:creator>
<dc:creator>Schneider, A.</dc:creator>
<dc:creator>Ginno, P. A.</dc:creator>
<dc:creator>Schneider, N.</dc:creator>
<dc:creator>Koch, M.-L.</dc:creator>
<dc:creator>Gerstung, M.</dc:creator>
<dc:creator>Stegle, O.</dc:creator>
<dc:creator>Turner, J. M. A.</dc:creator>
<dc:creator>Heard, E.</dc:creator>
<dc:creator>Odom, D. T.</dc:creator>
<dc:date>2023-12-05</dc:date>
<dc:identifier>doi:10.1101/2023.12.04.569933</dc:identifier>
<dc:title><![CDATA[Four-Core Genotypes mice harbour a 3.2MB X-Y translocation that perturbs Tlr7 dosage]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.10.570962v1?rss=1">
<title>
<![CDATA[
The Legionella collagen-like protein employs a unique binding mechanism for the recognition of host glycosaminoglycans 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.10.570962v1?rss=1"
</link>
<description><![CDATA[
Bacterial adhesion is a fundamental process which enables colonisation of niche environments and is key for infection. However, in Legionella pneumophila, the causative agent of Legionnaires disease, these processes are not well understood. The Legionella collagen-like protein (Lcl) is an extracellular peripheral membrane protein that recognises sulphated glycosaminoglycans (GAGs) on the surface of eukaryotic cells, but also stimulates bacterial aggregation in response to divalent cations. Here we report the crystal structure of the Lcl C-terminal domain (Lcl-CTD) and present a model for intact Lcl. Our data reveal that Lcl-CTD forms an unusual dynamic trimer arrangement with a positively charged external surface and a negatively charged solvent exposed internal cavity. Through Molecular Dynamics (MD) simulations, we show how the GAG chondroitin-4-sulphate associates with the Lcl-CTD surface via unique binding modes. Our findings show that Lcl homologs are present across both the Pseudomonadota and Fibrobacterota-Chlorobiota-Bacteroidota phyla and suggest that Lcl may represent a versatile carbohydrate binding mechanism.
]]></description>
<dc:creator>Rehman, S.</dc:creator>
<dc:creator>Antonovic, A.</dc:creator>
<dc:creator>McIntire, I.</dc:creator>
<dc:creator>Zheng, H.</dc:creator>
<dc:creator>Cleaver, L.</dc:creator>
<dc:creator>Adams, C.</dc:creator>
<dc:creator>Portlock, T.</dc:creator>
<dc:creator>Richardson, K.</dc:creator>
<dc:creator>Shaw, R.</dc:creator>
<dc:creator>Oregioni, A.</dc:creator>
<dc:creator>Mastroianni, G.</dc:creator>
<dc:creator>Whittaker, S.</dc:creator>
<dc:creator>Kelly, G.</dc:creator>
<dc:creator>Fornili, A.</dc:creator>
<dc:creator>Cianciotto, N.</dc:creator>
<dc:creator>Garnett, J.</dc:creator>
<dc:date>2023-12-10</dc:date>
<dc:identifier>doi:10.1101/2023.12.10.570962</dc:identifier>
<dc:title><![CDATA[The Legionella collagen-like protein employs a unique binding mechanism for the recognition of host glycosaminoglycans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.10.570980v1?rss=1">
<title>
<![CDATA[
Brain-wide activity-identity mapping of neural networks associated with prosocial motivation in rats 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.10.570980v1?rss=1"
</link>
<description><![CDATA[
A prosocial response to others in distress is an important driver of behavior across social species. To investigate the neurobiological mechanism leading to prosocial behavior, we use a helping behavior test wherein rats may release a trapped conspecific by opening a restrainer door. To ensure rats were not acting for social interaction, a separation divider prevented post-release contact ("separated" test). Despite the divider, most rats consistently opened the restrainer, demonstrating prosocial motivation. Brain-wide c-Fos analysis conducted via our opensource software "Brainways", revealed activity in empathy-related regions, including the anterior cingulate and insular cortices. Nucleus accumbens activity, previously recorded during helping, was not significant in the "separated" test. Chemogenetic manipulations of the accumbens did not prevent helping, suggesting that its activity reflects contact seeking. Mapping of oxytocin and dopamine receptors on active cells revealed region-specific recruitment of these subpopulations, depending on the social context. Network connectivity analysis highlights context-dependent functional subcircuits.
]]></description>
<dc:creator>Ruzal, K.</dc:creator>
<dc:creator>Trachtenberg, E.</dc:creator>
<dc:creator>Kantor, B.</dc:creator>
<dc:creator>Flumin, H.</dc:creator>
<dc:creator>Roemer, A.</dc:creator>
<dc:creator>Crespo, A.</dc:creator>
<dc:creator>Kohl, J.</dc:creator>
<dc:creator>Ben-Ami Bartal, I.</dc:creator>
<dc:date>2023-12-11</dc:date>
<dc:identifier>doi:10.1101/2023.12.10.570980</dc:identifier>
<dc:title><![CDATA[Brain-wide activity-identity mapping of neural networks associated with prosocial motivation in rats]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.12.571217v1?rss=1">
<title>
<![CDATA[
A cyclic peptide toolkit reveals mechanistic principles of peptidylarginine deiminase IV (PADI4) regulation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.12.571217v1?rss=1"
</link>
<description><![CDATA[
Peptidylarginine deiminase IV (PADI4) deregulation promotes the development of autoimmunity, cancer, atherosclerosis and age-related tissue fibrosis. Genetic or pharmacological PADI4 inhibition is therapeutically efficacious in mice, but no clinically relevant inhibitors currently exist. PADI4 additionally mediates immune responses and cellular reprogramming, although the full extent of its physiological roles is unexplored. Despite detailed molecular knowledge of PADI4 activation in vitro, we lack understanding of its regulation within cells, largely due to lack of appropriate systems and tools. Here, we developed and applied a set of potent and selective PADI4 modulators. Using the mRNA- display-based RaPID system, we screened >1012 cyclic peptides for high-affinity, conformation-selective binders. We report PADI4_3, an inhibitor specific for the active conformation of PADI4; PADI4_7, an inert binder, which we functionalised for the isolation and study of cellular PADI4; and PADI4_11, a first-in-class activator. Using a newly developed method for the quantification of cellular PADI4 activity, we show that PADI4_3 and PADI4_11 are effective in cells. Structural studies with PADI4_11 reveal an allosteric binding mode that may reflect the mechanism that promotes cellular PADI4 activation. This work offers new understanding of PADI4 regulation and a toolkit for the study and modulation of PADI4 across physiological and pathophysiological contexts.
]]></description>
<dc:creator>Bertran, M. T.</dc:creator>
<dc:creator>Walmsley, R.</dc:creator>
<dc:creator>Cummings, T. F. M.</dc:creator>
<dc:creator>Aramburu, I.</dc:creator>
<dc:creator>Benton, D. J.</dc:creator>
<dc:creator>Assalaarachchi, J.</dc:creator>
<dc:creator>Chasampalioti, M.</dc:creator>
<dc:creator>Swanton, T.</dc:creator>
<dc:creator>Joshi, D.</dc:creator>
<dc:creator>Federico, S.</dc:creator>
<dc:creator>Okkenhaug, H.</dc:creator>
<dc:creator>Yu, L.</dc:creator>
<dc:creator>Oxley, D.</dc:creator>
<dc:creator>Walker, S.</dc:creator>
<dc:creator>Papayannopoulos, V.</dc:creator>
<dc:creator>Suga, H.</dc:creator>
<dc:creator>Christophorou, M. A.</dc:creator>
<dc:creator>Walport, L. J.</dc:creator>
<dc:date>2023-12-12</dc:date>
<dc:identifier>doi:10.1101/2023.12.12.571217</dc:identifier>
<dc:title><![CDATA[A cyclic peptide toolkit reveals mechanistic principles of peptidylarginine deiminase IV (PADI4) regulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.19.572445v1?rss=1">
<title>
<![CDATA[
Live-imaging reveals Coordinated Cell Migration and Cardiac Fate Determination during Mammalian Gastrulation. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.19.572445v1?rss=1"
</link>
<description><![CDATA[
Heart development involves the specification of cardiac progenitors at distinct stages and locations. Using live-imaging of mouse embryos between gastrulation and heart tube formation, we tracked individual mesodermal cells and reconstructed their lineage trees for up to five cell divisions. We found independent unipotent progenitors emerging at specific times, contributing exclusively to either left ventricle/atrioventricular canal (LV/AVC) or atrial myocytes. LV/AVC progenitors differentiated early to form the cardiac crescent, while atrial progenitors later generated the heart tubes inflow tract during morphogenesis. We also identified short-lived bipotent progenitors with broad potential, illustrating early developmental plasticity. Sister cells from bipotent progenitors displayed greater dispersion and more diverse migratory trajectories within the anterior mesoderm than those from unipotent progenitors. Bipotent progenitors contributing to extraembryonic mesoderm (ExEm) exhibited the fastest and most dispersed migrations, whereas those giving rise to endocardial, LV/AVC, and pericardial cells showed a more gradual divergence, with late-stage behavioural shifts: endocardial cells increased in speed, while pericardial cells slowed relative to LV/AVC cells. Together the data reveal the regulation of individual cell directionality and cardiac fate allocation within the seemingly unorganised migratory pattern of mesoderm cells.
]]></description>
<dc:creator>Abukar, S.</dc:creator>
<dc:creator>Embacher, P.</dc:creator>
<dc:creator>Cicarrelli, A.</dc:creator>
<dc:creator>Varsani-Brown, S.</dc:creator>
<dc:creator>Dean, J. A.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Ivanovitch, k.</dc:creator>
<dc:date>2023-12-19</dc:date>
<dc:identifier>doi:10.1101/2023.12.19.572445</dc:identifier>
<dc:title><![CDATA[Live-imaging reveals Coordinated Cell Migration and Cardiac Fate Determination during Mammalian Gastrulation.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.20.572522v1?rss=1">
<title>
<![CDATA[
Xylosyltransferase Bump-and-hole Engineering to Chemically Manipulate Proteoglycans in Mammalian Cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.20.572522v1?rss=1"
</link>
<description><![CDATA[
Mammalian cells orchestrate signalling through interaction events on their surfaces. Proteoglycans are an intricate part of these interactions, carrying large glycosaminoglycan polysaccharides that recruit signalling molecules. Despite their importance in development, cancer and neurobiology, a relatively small number of proteoglycans have been identified. In addition to the complexity of glycan extension, biosynthetic redundancy in the first protein glycosylation step by two xylosyltransferase isoenzymes XT1 and XT2 complicates annotation of proteoglycans. Here, we develop a chemical genetic strategy that manipulates the glycan attachment site of cellular proteoglycans. By employing a tactic termed bump- and-hole engineering, we engineer the two isoenzymes XT1 and XT2 to specifically transfer a chemically modified xylose analogue to target proteins. The chemical modification contains a bioorthogonal tag, allowing the ability to visualise and profile target proteins modified by both transferases in mammalian cells. The versatility of our approach allows pinpointing glycosylation sites by tandem mass spectrometry, and exploiting the chemical handle to manufacture proteoglycans with defined GAG chains for cellular applications. Engineered XT enzymes permit a view into proteoglycan biology that is orthogonal to conventional techniques in biochemistry.
]]></description>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Di Vagno, L.</dc:creator>
<dc:creator>Ni Cheallaigh, A.</dc:creator>
<dc:creator>Sammon, D.</dc:creator>
<dc:creator>Briggs, D. C.</dc:creator>
<dc:creator>Chung, N.</dc:creator>
<dc:creator>Chang, V.</dc:creator>
<dc:creator>Mahoney, K. E.</dc:creator>
<dc:creator>Cioce, A.</dc:creator>
<dc:creator>Murphy, L. D.</dc:creator>
<dc:creator>Chen, Y.-H.</dc:creator>
<dc:creator>Narimatsu, Y.</dc:creator>
<dc:creator>Miller, R. L.</dc:creator>
<dc:creator>Willems, L. I.</dc:creator>
<dc:creator>Malaker, S. A.</dc:creator>
<dc:creator>Miller, G. J.</dc:creator>
<dc:creator>Hohenester, E.</dc:creator>
<dc:creator>Schumann, B.</dc:creator>
<dc:date>2023-12-20</dc:date>
<dc:identifier>doi:10.1101/2023.12.20.572522</dc:identifier>
<dc:title><![CDATA[Xylosyltransferase Bump-and-hole Engineering to Chemically Manipulate Proteoglycans in Mammalian Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.19.572309v1?rss=1">
<title>
<![CDATA[
The V-ATPase/ATG16L1 axis is controlled by the V1H subunit 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.19.572309v1?rss=1"
</link>
<description><![CDATA[
Defects in organellar acidification indicate compromised or infected compartments. Recruitment of the autophagy-related ATG16L1 complex to pathologically de-acidified compartments targets ubiquitin-like ATG8 molecules to perturbed membranes. How this process is coupled to pH gradient disruption is unclear. Here, we reveal a direct role for the V1H subunit of the V-ATPase proton pump in recruiting ATG16L1. The interaction between V1H and ATG16L1 occurs within assembled V-ATPases, but not dissociated V1 complexes. This selectivity allows recruitment to be coupled to changes in V-ATPase assembly that follow pH dissipation. Cells lacking V1H undergo canonical macroautophagy but are unable to recruit ATG16L1 in response to influenza infection or ionophore drugs. We identify a loop within V1H that mediates ATG16L1 binding, which is absent in a neuronal isoform of V1H. Thus, V1H controls ATG16L1 recruitment in response to proton gradient dissipation, suggesting that the V-ATPase acts autonomously as a cell-intrinsic damage sensor.
]]></description>
<dc:creator>Timimi, L.</dc:creator>
<dc:creator>Wrobel, A. G.</dc:creator>
<dc:creator>Chiduza, G. N.</dc:creator>
<dc:creator>Maslen, S. L.</dc:creator>
<dc:creator>Torres-Mendez, A.</dc:creator>
<dc:creator>Montaner, B.</dc:creator>
<dc:creator>Davis, C.</dc:creator>
<dc:creator>Skehel, M.</dc:creator>
<dc:creator>Rubinstein, J. L.</dc:creator>
<dc:creator>Schreiber, A. L.</dc:creator>
<dc:creator>Beale, R.</dc:creator>
<dc:date>2023-12-19</dc:date>
<dc:identifier>doi:10.1101/2023.12.19.572309</dc:identifier>
<dc:title><![CDATA[The V-ATPase/ATG16L1 axis is controlled by the V1H subunit]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.16.572007v1?rss=1">
<title>
<![CDATA[
Imaging the master regulator of the antioxidant response in non-small cell lung cancer with positron emission tomography 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.16.572007v1?rss=1"
</link>
<description><![CDATA[
Mutations in the NRF2-KEAP1 pathway are common in non-small cell lung cancer (NSCLC) and confer broad-spectrum therapeutic resistance, leading to poor outcomes. The cystine/glutamate antiporter, system xc-, is one of the >200 cytoprotective proteins controlled by NRF2, which can be non-invasively imaged by (S)-4-(3-18F-fluoropropyl)--glutamate ([18F]FSPG) positron emission tomography (PET). Through genetic and pharmacologic manipulation, we show that [18F]FSPG provides a sensitive and specific marker of NRF2 activation in advanced preclinical models of NSCLC. We validate imaging readouts with metabolomic measurements of system xc- activity and their coupling to intracellular glutathione concentration. A redox gene signature was measured in patients from the TRACERx 421 cohort, suggesting an opportunity for patient stratification prior to imaging. Furthermore, we reveal that system xc- is a metabolic vulnerability that can be therapeutically targeted for sustained tumour growth suppression in aggressive NSCLC. Our results establish [18F]FSPG as predictive marker of therapy resistance in NSCLC and provide the basis for the clinical evaluation of both imaging and therapeutic agents that target this important antioxidant pathway.
]]></description>
<dc:creator>Greenwood, H. E.</dc:creator>
<dc:creator>Edwards, R. S.</dc:creator>
<dc:creator>Tyrrell, W. E.</dc:creator>
<dc:creator>Barber, A. R.</dc:creator>
<dc:creator>Baark, F.</dc:creator>
<dc:creator>Tanc, M.</dc:creator>
<dc:creator>Khalil, E.</dc:creator>
<dc:creator>Falzone, A.</dc:creator>
<dc:creator>Ward, N. P.</dc:creator>
<dc:creator>DeBlasi, J. M.</dc:creator>
<dc:creator>Torrente, L.</dc:creator>
<dc:creator>Pearce, D. R.</dc:creator>
<dc:creator>Firth, G.</dc:creator>
<dc:creator>Smith, L. M.</dc:creator>
<dc:creator>Vilhelmsson Timmermand, O.</dc:creator>
<dc:creator>Huebner, A.</dc:creator>
<dc:creator>George, M. E.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Hynds, R. E.</dc:creator>
<dc:creator>DeNicola, G. M.</dc:creator>
<dc:creator>Witney, T. H.</dc:creator>
<dc:date>2023-12-17</dc:date>
<dc:identifier>doi:10.1101/2023.12.16.572007</dc:identifier>
<dc:title><![CDATA[Imaging the master regulator of the antioxidant response in non-small cell lung cancer with positron emission tomography]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.18.572148v1?rss=1">
<title>
<![CDATA[
RASP: Optimal single fluorescent puncta detection in complex cellular backgrounds 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.18.572148v1?rss=1"
</link>
<description><![CDATA[
Super-resolution and single-molecule microscopy are increasingly applied to complex biological systems. A major challenge of this approach is that fluorescent puncta must be detected in the low signal, high noise, heterogeneous background environments of cells and tissue. We present RASP, Radiality Analysis of Single Puncta, a bioimaging-segmentation method that solves this problem. RASP removes false positive puncta that other analysis methods detect, and detects features over a broad range of spatial scales: from single proteins to complex cell phenotypes. RASP outperforms the state-of-the-art in precision and speed, using image gradients to separate Gaussian-shaped objects from background. We demonstrate RASPs power by showing it can extract spatial correlations between microglia, neurons, and -synuclein oligomers in the human brain. This sensitive, computationally efficient approach enables fluorescent puncta and cellular features to be distinguished in cellular and tissue environments with a sensitivity down to the level of the single protein.
]]></description>
<dc:creator>Fu, B.</dc:creator>
<dc:creator>Brock, E. E.</dc:creator>
<dc:creator>Breiter, J. C.</dc:creator>
<dc:creator>Tian, R.</dc:creator>
<dc:creator>Andrews, R.</dc:creator>
<dc:creator>Toomey, C. E.</dc:creator>
<dc:creator>Lachica, J.</dc:creator>
<dc:creator>Lashley, T.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:creator>Wood, N. W.</dc:creator>
<dc:creator>Vendruscolo, M.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Weiss, L. E.</dc:creator>
<dc:creator>Beckwith, J. S.</dc:creator>
<dc:creator>Lee, S. F.</dc:creator>
<dc:date>2023-12-18</dc:date>
<dc:identifier>doi:10.1101/2023.12.18.572148</dc:identifier>
<dc:title><![CDATA[RASP: Optimal single fluorescent puncta detection in complex cellular backgrounds]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.20.572534v1?rss=1">
<title>
<![CDATA[
Shieldin and CST co-orchestrate DNA polymerase-dependent tailed-end joining reactions independently of 53BP1-governed repair pathway choice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.20.572534v1?rss=1"
</link>
<description><![CDATA[
53BP1 regulates DNA end-joining in lymphocytes, diversifying immune antigen receptors. This involves nucleosome-bound 53BP1 at DNA double-stranded breaks (DSBs) recruiting RIF1 and shieldin, a poorly understood DNA-binding complex. The 53BP1-RIF1-shieldin axis is pathological in BRCA1-mutated cancers, blocking homologous recombination (HR) and driving illegitimate non-homologous end-joining (NHEJ). However, how this axis regulates DNA end-joining and HR suppression remains unresolved.

We investigated shieldin and its interplay with CST, a complex recently implicated in 53BP1-dependent activities. Immunophenotypically, mice lacking shieldin or CST are equivalent, with class-switch recombination co-reliant on both complexes. ATM-dependent DNA damage signalling underpins this cooperation, inducing physical interactions between these complexes that reveal shieldin as a DSB-responsive CST adaptor. Furthermore, DNA polymerase {zeta} functions downstream of shieldin, establishing DNA fill-in synthesis as the physiological function of shieldin-CST. Lastly, 53BP1 suppresses HR and promotes NHEJ in BRCA1-deficient mice and cells independently of shieldin. These findings showcase the resilience of the 53BP1 pathway, achieved through the collaboration of chromatin-bound 53BP1 complexes and DNA end-processing effector proteins.
]]></description>
<dc:creator>King, A.</dc:creator>
<dc:creator>Reichl, P.</dc:creator>
<dc:creator>Metson, J. S.</dc:creator>
<dc:creator>Parker, R.</dc:creator>
<dc:creator>Munro, D.</dc:creator>
<dc:creator>Oliveira, C.</dc:creator>
<dc:creator>Becker, J. R.</dc:creator>
<dc:creator>Biggs, D.</dc:creator>
<dc:creator>Preece, C.</dc:creator>
<dc:creator>Davies, B.</dc:creator>
<dc:creator>Chapman, J. R.</dc:creator>
<dc:date>2023-12-21</dc:date>
<dc:identifier>doi:10.1101/2023.12.20.572534</dc:identifier>
<dc:title><![CDATA[Shieldin and CST co-orchestrate DNA polymerase-dependent tailed-end joining reactions independently of 53BP1-governed repair pathway choice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.23.573207v1?rss=1">
<title>
<![CDATA[
Oncogenic PIK3CA corrupts growth factor signaling specificity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.23.573207v1?rss=1"
</link>
<description><![CDATA[
Pathological activation of the PI3K/AKT pathway is among the most frequent defects in human cancer and is also the cause of rare overgrowth disorders. Yet, there is currently no systematic understanding of the quantitative flow of information within PI3K/AKT signaling and how it is perturbed by disease-causing mutations. Here, we develop scalable, single-cell approaches for systematic analyses of signal processing within the PI3K pathway, enabling precise calculations of its information transfer for different growth factors. Using genetically-engineered human cell models with allele dose-dependent expression of PIK3CAH1047R, we show that this oncogene is not a simple, constitutive pathway activator but a context-dependent modulator of extracellular signal transfer. PIK3CAH1047Rreduces information transmission downstream of IGF1 while selectively enhancing EGF-induced signaling and transcriptional responses. This leads to a gross reduction in signaling specificity, akin to "blurred" signal perception. The associated increase in signaling heterogeneity promotes phenotypic diversity in a human cervical cancer cell line model and in human induced pluripotent stem cells. Collectively, these findings and the accompanying methodological advances lay the foundations for a systematic mapping of the quantitative mechanisms of PI3K/AKT-dependent signal processing and phenotypic control in health and disease.

One-sentence summarySingle-cell signaling and information theoretic analyses reveal that oncogenic PI3K/AKT activation leads to a gross reduction in signaling specificity, context-dependent EGF response amplification as well as increased phenotypic heterogeneity.
]]></description>
<dc:creator>Madsen, R. R.</dc:creator>
<dc:creator>Le Marois, A.</dc:creator>
<dc:creator>Voliotis, M.</dc:creator>
<dc:creator>Yin, S.</dc:creator>
<dc:creator>Sufi, J.</dc:creator>
<dc:creator>Qin, X.</dc:creator>
<dc:creator>Zhao, S. J.</dc:creator>
<dc:creator>Gorczynska, J.</dc:creator>
<dc:creator>Morelli, D.</dc:creator>
<dc:creator>Davidson, L.</dc:creator>
<dc:creator>Sahai, E.</dc:creator>
<dc:creator>Korolchuk, V. I.</dc:creator>
<dc:creator>Tape, C. J.</dc:creator>
<dc:creator>Vanhaesebroeck, B.</dc:creator>
<dc:date>2023-12-23</dc:date>
<dc:identifier>doi:10.1101/2023.12.23.573207</dc:identifier>
<dc:title><![CDATA[Oncogenic PIK3CA corrupts growth factor signaling specificity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.04.574237v1?rss=1">
<title>
<![CDATA[
Glutaminase as a metabolic target of choice to counter acquired resistance to Palbociclib by colorectal cancer cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.04.574237v1?rss=1"
</link>
<description><![CDATA[
Several mechanisms of resistance of cancer cells to cyclin-dependent kinase inhibitors (CDKi) have been identified, including the upregulation of metabolic regulators such as glutaminase. However, whether such mechanisms and targets are optimal has not been determined. Here, we have systematically analyzed metabolic reprogramming in colorectal cancer cells exposed to Palbociclib, a CDKi selectively targeting CDK4/6, or Telaglenestat, a selective glutaminase inhibitor. Through multiple approaches, we show that Palbociclib and Telaglenestat elicit complementary metabolic responses and are thus uniquely suited to counter the metabolic reprogramming induced by the reciprocal drug. As such, while Palbociclib induced reduced tumor growth in vivo, and Telaglenestat did not show a significant effect, the drug combination displayed a strong synergistic effect on tumor growth. Likewise, initial responses to Palbociclib were followed by signs of adaptation and resistance, which were prevented by combining Palbociclib with Telaglenestat. In conclusion, combination with Telaglenestat optimally forestalls acquired resistance to Palbociclib in cancer cells.
]]></description>
<dc:creator>Tarrado-Castellarnau, M.</dc:creator>
<dc:creator>Foguet, C.</dc:creator>
<dc:creator>Tarrago-Celada, J.</dc:creator>
<dc:creator>Palobart, M.</dc:creator>
<dc:creator>Hernandez-Carro, C.</dc:creator>
<dc:creator>Perarnau, J.</dc:creator>
<dc:creator>Zodda, E.</dc:creator>
<dc:creator>Polat, I. H.</dc:creator>
<dc:creator>Marin, S.</dc:creator>
<dc:creator>Suarez-Bonnet, A.</dc:creator>
<dc:creator>Lozano, J. J.</dc:creator>
<dc:creator>Yuneva, M.</dc:creator>
<dc:creator>Thomson, T. M.</dc:creator>
<dc:creator>Cascante, M.</dc:creator>
<dc:date>2024-01-05</dc:date>
<dc:identifier>doi:10.1101/2024.01.04.574237</dc:identifier>
<dc:title><![CDATA[Glutaminase as a metabolic target of choice to counter acquired resistance to Palbociclib by colorectal cancer cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.05.574362v1?rss=1">
<title>
<![CDATA[
Deciphering the diversity and sequence of extracellular matrix and cellular spatial patterns in lung adenocarcinoma using topological data analysis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.05.574362v1?rss=1"
</link>
<description><![CDATA[
Extracellular matrix (ECM) organization influences cancer development and progression. It modulates the invasion of cancer cells and can hinder the access of immune cells to cancer cells. Effective quantification of ECM architecture and its relationship to the position of different cell types is, therefore, important when investigating the role of ECM in cancer development. Using topological data analysis (TDA), particularly persistent homology and Dowker persistent homology, we develop a novel analysis pipeline for quantifying ECM architecture, spatial patterns of cell positions, and the spatial relationships between distinct constituents of the tumour microenvironment. We apply the pipeline to 44 surgical specimens of lung adenocarcinoma from the lung TRACERx study stained with picrosirius red and haematoxylin. We show that persistent homology effectively encodes the architectural features of the tumour microenvironment. Inference using pseudo-time analysis and spatial mapping to centimetre scale tissues suggests a gradual and progressive route of change in ECM architecture, with two different end states. Dowker persistent homology enables the analysis of spatial relationship between any pair of constituents of the tumour microenvironment, such as ECM, cancer cells, and leukocytes. We use Dowker persistent homology to quantify the spatial segregation of cancer and immune cells over different length scales. A combined analysis of both topological and non-topological features of the tumour microenvironment indicates that progressive changes in the ECM are linked to increased immune exclusion and reduced oxidative metabolism.
]]></description>
<dc:creator>Yoon, I. H. R.</dc:creator>
<dc:creator>Jenkins, R.</dc:creator>
<dc:creator>Colliver, E.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Novo, D.</dc:creator>
<dc:creator>Moore, D.</dc:creator>
<dc:creator>Ramsden, Z.</dc:creator>
<dc:creator>Rullan, A.</dc:creator>
<dc:creator>Fu, X.</dc:creator>
<dc:creator>Yuan, Y.</dc:creator>
<dc:creator>Harrington, H. A.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Byrne, H. M.</dc:creator>
<dc:creator>Sahai, E.</dc:creator>
<dc:date>2024-01-07</dc:date>
<dc:identifier>doi:10.1101/2024.01.05.574362</dc:identifier>
<dc:title><![CDATA[Deciphering the diversity and sequence of extracellular matrix and cellular spatial patterns in lung adenocarcinoma using topological data analysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.03.574108v1?rss=1">
<title>
<![CDATA[
Single-cell transcriptional dynamics in a living vertebrate 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.03.574108v1?rss=1"
</link>
<description><![CDATA[
The ability to quantify transcriptional dynamics in individual cells via live imaging has revolutionized our understanding of gene regulation. However, such measurements are lacking in the context of vertebrate embryos. We addressed this deficit by applying MS2-MCP mRNA labeling to the quantification of transcription in zebrafish, a model vertebrate. We developed a platform of transgenic organisms, light sheet fluorescence microscopy, and optimized image analysis that enables visualization and quantification of MS2 reporters. With these tools, we obtained single-cell, real-time measurements of transcriptional dynamics of the segmentation clock. Our measurements challenge the traditional view of smooth clock oscillations and instead suggest a model of discrete transcriptional bursts that are organized in space and time. Together, these results highlight how measuring single-cell transcriptional activity in the context of vertebrate organisms can reveal unexpected features of gene regulation and how this data can fuel the dialogue between theory and experiment.
]]></description>
<dc:creator>Eck, E.</dc:creator>
<dc:creator>Moretti, B.</dc:creator>
<dc:creator>Schlomann, B. H.</dc:creator>
<dc:creator>Bragantini, J.</dc:creator>
<dc:creator>Lange, M.</dc:creator>
<dc:creator>Zhao, X.</dc:creator>
<dc:creator>VijayKumar, S.</dc:creator>
<dc:creator>Valentin, G.</dc:creator>
<dc:creator>Loureiro, C.</dc:creator>
<dc:creator>Soroldoni, D.</dc:creator>
<dc:creator>Royer, L. A.</dc:creator>
<dc:creator>Oates, A. C.</dc:creator>
<dc:creator>Garcia, H. H.</dc:creator>
<dc:date>2024-01-05</dc:date>
<dc:identifier>doi:10.1101/2024.01.03.574108</dc:identifier>
<dc:title><![CDATA[Single-cell transcriptional dynamics in a living vertebrate]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.10.575016v1?rss=1">
<title>
<![CDATA[
Cell Cycle Regulation has Shaped Budding Yeast Replication Origin Structure and Function 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.10.575016v1?rss=1"
</link>
<description><![CDATA[
Eukaryotic DNA replication initiates from multiple genomic loci known as origins. At budding yeast origins like ARS1, a double hexamer (DH) of the MCM replicative helicase is assembled by Origin Recognition Complex (ORC), Cdc6 and Cdt1 via sequential hexamer loading from two opposed ORC binding sites. Cyclin Dependent Kinase (CDK) inhibits DH assembly, which prevents re-replication by restricting helicase loading to G1 phase. Here we show that an intrinsically disordered region (IDR) in the Orc2 subunit promotes interaction between ORC and the first loaded, closed-ring MCM hexamer (the MO intermediate); CDK phosphorylation of this IDR blocks MO formation and DH assembly. We show that MO functions by stabilising ORC at the lower affinity binding sites required for second hexamer loading. Origins comprising two high affinity ORC sites can assemble DH efficiently without MO by independently loading single hexamers; these origins escape CDK inhibition in vitro and in vivo. Our work reveals mechanistic plasticity in MCM loading with implications for understanding how CDK regulation has shaped yeast origin evolution and how natural origins might escape cell cycle regulation. We also identify key steps common to loading pathways, with implications for understanding how MCM is loaded in other eukaryotes.
]]></description>
<dc:creator>Diffley, J.</dc:creator>
<dc:creator>Lim, C. T.</dc:creator>
<dc:creator>Miller, T. C.</dc:creator>
<dc:creator>Tan, K. W.</dc:creator>
<dc:creator>Talele, s.</dc:creator>
<dc:creator>Early, A.</dc:creator>
<dc:creator>East, P.</dc:creator>
<dc:creator>Sanchez, H.</dc:creator>
<dc:creator>Dekker, N. H.</dc:creator>
<dc:creator>Costa, A.</dc:creator>
<dc:date>2024-01-10</dc:date>
<dc:identifier>doi:10.1101/2024.01.10.575016</dc:identifier>
<dc:title><![CDATA[Cell Cycle Regulation has Shaped Budding Yeast Replication Origin Structure and Function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.11.575224v1?rss=1">
<title>
<![CDATA[
Mixed Alkyl/Aryl Phosphonates Identify Metabolic Serine Hydrolases as Antimalarial Targets 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.11.575224v1?rss=1"
</link>
<description><![CDATA[
Malaria, caused by Plasmodium falciparum, remains a significant health burden. A barrier for developing anti-malarial drugs is the ability of the parasite to rapidly generate resistance. We demonstrated that Salinipostin A (SalA), a natural product, kills parasites by inhibiting multiple lipid metabolizing serine hydrolases, a mechanism with a low propensity for resistance. Given the difficulty of employing natural products as therapeutic agents, we synthesized a library of lipidic mixed alkyl/aryl phosphonates as bioisosteres of SalA. Two constitutional isomers exhibited divergent anti-parasitic potencies which enabled identification of therapeutically relevant targets. We also confirm that this compound kills parasites through a mechanism that is distinct from both SalA and the pan-lipase inhibitor, Orlistat. Like SalA, our compound induces only weak resistance, attributable to mutations in a single protein involved in multidrug resistance. These data suggest that mixed alkyl/aryl phosphonates are a promising, synthetically tractable anti-malarials with a low-propensity to induce resistance.
]]></description>
<dc:creator>Bennett, J. M.</dc:creator>
<dc:creator>Narwal, S. K.</dc:creator>
<dc:creator>Kabeche, S.</dc:creator>
<dc:creator>Abegg, D.</dc:creator>
<dc:creator>Hackett, F.</dc:creator>
<dc:creator>Yeo, T.</dc:creator>
<dc:creator>Li, V.</dc:creator>
<dc:creator>Muir, R. K.</dc:creator>
<dc:creator>Faucher, F. F.</dc:creator>
<dc:creator>Lovell, S.</dc:creator>
<dc:creator>Blackman, M.</dc:creator>
<dc:creator>Adibekian, A.</dc:creator>
<dc:creator>Yeh, E.</dc:creator>
<dc:creator>Fidock, D. A.</dc:creator>
<dc:creator>Bogyo, M.</dc:creator>
<dc:date>2024-01-11</dc:date>
<dc:identifier>doi:10.1101/2024.01.11.575224</dc:identifier>
<dc:title><![CDATA[Mixed Alkyl/Aryl Phosphonates Identify Metabolic Serine Hydrolases as Antimalarial Targets]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.11.575158v1?rss=1">
<title>
<![CDATA[
Targeting the Plasmodium falciparum UCHL3 ubiquitin hydrolase using chemically constrained peptides 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.11.575158v1?rss=1"
</link>
<description><![CDATA[
The ubiquitin-proteasome system is essential to all eukaryotes and has been shown to be critical to parasite survival as well, including Plasmodium falciparum, the causative agent of the deadliest form of malarial disease. Despite the central role of the ubiquitin-proteasome pathway to parasite viability across its entire life-cycle, specific inhibitors targeting the individual enzymes mediating ubiquitin attachment and removal do not currently exist. The ability to disrupt P. falciparum growth at multiple developmental stages is particularly attractive as this could potentially prevent both disease pathology, caused by asexually dividing parasites, as well as transmission which is mediated by sexually differentiated parasites. The deubiquitinating enzyme PfUCHL3 is an essential protein, transcribed across both human and mosquito developmental stages. PfUCHL3 is considered hard to drug by conventional methods given the high level of homology of its active site to human UCHL3 as well as to other UCH domain enzymes. Here, we apply the RaPID mRNA display technology and identify constrained peptides capable of binding to PfUCHL3 with nanomolar affinities. The two lead peptides were found to selectively inhibit the deubiquitinase activity of PfUCHL3 versus HsUCHL3. NMR spectroscopy revealed that the peptides do not act by binding to the active site but instead block binding of the ubiquitin substrate. We demonstrate that this approach can be used to target essential protein-protein interactions within the Plasmodium ubiquitin pathway, enabling the application of chemically constrained peptides as a novel class of anti-malarial therapeutics.
]]></description>
<dc:creator>King, H. R.</dc:creator>
<dc:creator>Bycroft, M.</dc:creator>
<dc:creator>Nguyen, T.-B.</dc:creator>
<dc:creator>Kelly, G.</dc:creator>
<dc:creator>Vinogradov, A. A.</dc:creator>
<dc:creator>Rowling, P. J.</dc:creator>
<dc:creator>Stott, K.</dc:creator>
<dc:creator>Ascher, D. B.</dc:creator>
<dc:creator>Suga, H.</dc:creator>
<dc:creator>Itzhaki, L. S.</dc:creator>
<dc:creator>Artavanis-Tsakonas, K.</dc:creator>
<dc:date>2024-01-13</dc:date>
<dc:identifier>doi:10.1101/2024.01.11.575158</dc:identifier>
<dc:title><![CDATA[Targeting the Plasmodium falciparum UCHL3 ubiquitin hydrolase using chemically constrained peptides]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.14.575607v1?rss=1">
<title>
<![CDATA[
A scaleable inducible knockout systemfor studying essential gene function inthe malaria parasite 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.14.575607v1?rss=1"
</link>
<description><![CDATA[
The malaria parasite needs nearly half of its genes to propagate normally within red blood cells. Inducible ways to interfere with gene expression like the DiCre-lox system is necessary to study the function of these essential genes. However, the existing DiCre-lox strategy is not well-suited to be deployed at scale to study several genes simultaneously. To overcome this, we have developed SHIFTiKO (frameshift-based trackable inducible knockout), a novel scaleable strategy that uses short, easy-to-construct, barcoded repair templates to insert loxP sites around short regions in the target genes. Induced DiCre-mediated excision of the flanked region causes a frameshift mutation resulting in genetic ablation of gene function. Dual DNA barcodes inserted into each mutant enables verification of successful modification and induced excision at each locus and collective phenotyping of the mutants, not only across multiple replication cycles to assess growth fitness but also within a single cycle to identify the specific phenotypic impairment they exhibit. As a proof of concept, we have applied SHIFTiKO to screen the functions of malarial rhomboid proteases, successfully identifying their blood stage-specific essentiality. SHIFTiKO, thus offers a powerful platform to conduct inducible phenotypic screens to study essential gene function at scale in the malaria parasite.
]]></description>
<dc:creator>Ramaprasad, A.</dc:creator>
<dc:creator>Blackman, M.</dc:creator>
<dc:date>2024-01-15</dc:date>
<dc:identifier>doi:10.1101/2024.01.14.575607</dc:identifier>
<dc:title><![CDATA[A scaleable inducible knockout systemfor studying essential gene function inthe malaria parasite]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.14.575605v1?rss=1">
<title>
<![CDATA[
Quantifying spectral information about source separation in multisource odour plumes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.14.575605v1?rss=1"
</link>
<description><![CDATA[
Odours released by objects in natural environments can contain information about their spatial locations. In particular, the correlation of odour concentration fields produced by two spatially separated sources contains information about the distance between the sources. Mice are able to distinguish correlated and anti-correlated odour fluctuations at frequencies up to 40 Hz. Can this high-frequency acuity support odour source localization? Here we answer this question by quantifying the spatial information about source separation contained in the spectral constituents of correlations. We used computational fluid dynamics simulations of multisource plumes in two-dimensional chaotic flow environments to generate temporally complex, covarying odour concentration fields. By relating the correlation of these fields to the spectral decompositions of the associated odour concentration timeseries, and making simplifying assumptions about the statistics of these decompositions, we derived analytic expressions for the Fisher information contained in the spectral components of the correlations about source separation. We computed the Fisher information for a broad range of frequencies and source separations and found that high frequencies were more informative than low frequencies when sources were close relative to the sizes of the large eddies in the flow. We observed a qualitatively similar effect in an independent set of simulations with different geometry, but not for surrogate data with a similar power spectrum to our simulations but in which all frequencies were a priori equally informative. Our work suggests that the high-frequency acuity of the murine olfactory system may support high-resolution spatial localization of odour sources. We also provide a model of the distribution of the spectral components of correlations that is accurate over a broad range of frequencies and source separations. More broadly, our work establishes an approach for the quantification of the spatial information in odour concentration timeseries.
]]></description>
<dc:creator>Tootoonian, S.</dc:creator>
<dc:creator>True, A. C.</dc:creator>
<dc:creator>Crimaldi, J. P.</dc:creator>
<dc:creator>Schaefer, A. T.</dc:creator>
<dc:date>2024-01-15</dc:date>
<dc:identifier>doi:10.1101/2024.01.14.575605</dc:identifier>
<dc:title><![CDATA[Quantifying spectral information about source separation in multisource odour plumes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.15.575765v1?rss=1">
<title>
<![CDATA[
Combining RASG12C(ON) inhibitor with SHP2 inhibition sensitises immune excluded lung tumours to immune checkpoint blockade: a strategy for turning cold tumours hot 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.15.575765v1?rss=1"
</link>
<description><![CDATA[
Mutant selective drugs targeting the inactive, GDP-bound form of KRASG12C have been approved for use in lung cancer, but responses are short-lived due to rapid development of resistance. In this study we use a novel covalent tri-complex inhibitor, RMC-4998, that targets RASG12C in its active, GTP-bound form to investigate treatment of KRAS mutant lung cancer in various immune competent mouse models. While this RASG12C(ON) inhibitor was more potent than the KRASG12C(OFF) inhibitor adagrasib, rapid pathway reactivation was still observed. This could be delayed using combined treatment with a SHP2 inhibitor, RMC-4550, which not only impacted RAS pathway signalling within the tumour cells but also remodelled the tumour microenvironment (TME) to be less immunosuppressive and promoted interferon responses. In an inflamed, "hot", mouse model of lung cancer, RASG12C(ON) and SHP2 inhibitors in combination drive durable responses by suppressing tumour relapse and inducing development of immune memory, which can also be induced by combination of RASG12C(ON) and PD-1 inhibitors. In contrast, in an immune excluded, "cold", mouse model of lung cancer, combined RASG12C(ON) and SHP2 inhibition does not cause durable responses, but does sensitise tumours to immune checkpoint blockade, enabling efficient tumour rejection, accompanied by significant TME reorganization, including depletion of immunosuppressive innate immune cells and recruitment and activation of T and NK cells. These preclinical results demonstrate the potential of the combination of RASG12C(ON) inhibitors with SHP2 inhibitors to sensitize anti-PD-1 refractory tumours to immune checkpoint blockade by stimulating anti-tumour immunity as well as by targeting KRAS-driven proliferation in tumour cells.
]]></description>
<dc:creator>Anastasiou, P.</dc:creator>
<dc:creator>Moore, C.</dc:creator>
<dc:creator>Rana, S.</dc:creator>
<dc:creator>de Castro, A.</dc:creator>
<dc:creator>Tomaschko, M.</dc:creator>
<dc:creator>Boumelha, J.</dc:creator>
<dc:creator>Mugarza, E.</dc:creator>
<dc:creator>Blaj, C.</dc:creator>
<dc:creator>de Carne Trecesson, S.</dc:creator>
<dc:creator>Goldstone, R.</dc:creator>
<dc:creator>Smith, J.</dc:creator>
<dc:creator>Quintana, E.</dc:creator>
<dc:creator>Molina-Arcas, M.</dc:creator>
<dc:creator>Downward, J.</dc:creator>
<dc:date>2024-01-16</dc:date>
<dc:identifier>doi:10.1101/2024.01.15.575765</dc:identifier>
<dc:title><![CDATA[Combining RASG12C(ON) inhibitor with SHP2 inhibition sensitises immune excluded lung tumours to immune checkpoint blockade: a strategy for turning cold tumours hot]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.17.576008v1?rss=1">
<title>
<![CDATA[
Cryo-electron tomography reveals how COPII assembles on cargo-containing membranes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.17.576008v1?rss=1"
</link>
<description><![CDATA[
Proteins traverse the eukaryotic secretory pathway through membrane trafficking between organelles. The COPII coat mediates the anterograde transport of newly synthesised proteins from the endoplasmic reticulum, engaging cargoes with a wide range of size and biophysical properties. The native architecture of the COPII coat and how cargo might influence COPII carrier morphology remain poorly understood. Here, we have reconstituted COPII-coated membrane carriers using purified S. cerevisiae proteins and cell-derived microsomes as a native membrane source. Using cryo-electron tomography with subtomogram averaging, we demonstrate that the COPII coat binds cargo and forms largely spherical vesicles from native membranes. We reveal the architecture of the inner and outer coat layers and shed light on how spherical carriers are formed. Our results provide insights into the architecture and regulation of the COPII coat and advance our current understanding of how membrane curvature is generated.
]]></description>
<dc:creator>Pyle, E.</dc:creator>
<dc:creator>Zanetti, G.</dc:creator>
<dc:date>2024-01-17</dc:date>
<dc:identifier>doi:10.1101/2024.01.17.576008</dc:identifier>
<dc:title><![CDATA[Cryo-electron tomography reveals how COPII assembles on cargo-containing membranes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.15.575790v1?rss=1">
<title>
<![CDATA[
ImmCellTyper: an integrated computational pipeline for systematic mining of Mass Cytometry data to assist deep immune profiling 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.15.575790v1?rss=1"
</link>
<description><![CDATA[
Mass cytometry, also known as Cytometry by time-of-flight (CyTOF), is a cutting-edge high-dimensional technology for profiling marker expression at the single-cell level. This technology significantly advances clinical research in immune monitoring and the interrogation of immune cell populations. Nevertheless, the vast amount of data generated by CyTOF poses a daunting challenge for analysis. To address this, we describe ImmCellTyper (https://github.com/JingAnyaSun/ImmCellTyper), a novel and robust toolkit designed for CyTOF data analysis. The analytical framework incorporates an in-house developed semi-supervised clustering tool named BinaryClust, which first characterises main cell lineages, followed by in-depth interrogation for population of interest using unsupervised methods.

BinaryClust was benchmarked with existing clustering tools and demonstrated superior accuracy and speed across two datasets comprising around 4 million cells, performing as good as manual gating by human experts. Furthermore, this computational pipeline provides a variety of visualization and analytical tools spanning from quality control to differential analysis, which can be tailored to users specific needs, aiming to provide a one-stop solution for CyTOF data analysis. The general workflow consists of five key steps: 1) Batch effect evaluation and correction, 2) Data quality control and pre-processing, 3) Main cell lineage characterisation and quantification, 4) Extraction and in-depth investigation of cell type of interest; 5) Differential analysis of cell abundance and functional marker expression (supporting multiple study groups). Overall, ImmCellTyper integrates experts biological knowledge in a semi-supervised fashion to accurately deconvolute well-defined main cell lineages, while also preserving the potential of unsupervised approaches to discover novel cell subsets and providing a user-friendly toolset to remove the analytical barrier for high-dimensional immune profiling.
]]></description>
<dc:creator>Sun, J.</dc:creator>
<dc:creator>Choy, D.</dc:creator>
<dc:creator>Sompairac, N.</dc:creator>
<dc:creator>Jamshidi, S.</dc:creator>
<dc:creator>Mishto, M.</dc:creator>
<dc:creator>Kordasti, S.</dc:creator>
<dc:date>2024-01-17</dc:date>
<dc:identifier>doi:10.1101/2024.01.15.575790</dc:identifier>
<dc:title><![CDATA[ImmCellTyper: an integrated computational pipeline for systematic mining of Mass Cytometry data to assist deep immune profiling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.22.576655v1?rss=1">
<title>
<![CDATA[
Mechanism of chaperone coordination during cotranslational protein folding in bacteria 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.22.576655v1?rss=1"
</link>
<description><![CDATA[
Protein folding is assisted by molecular chaperones that bind nascent polypeptides during mRNA translation. Several structurally-distinct classes of chaperone promote de novo folding, suggesting that their activities are coordinated at the ribosome. We used biochemical reconstitution and structural proteomics to explore the molecular basis for cotranslational chaperone action in bacteria. We found that chaperone binding is disfavoured close to the ribosome, allowing folding to precede chaperone recruitment. Trigger factor subsequently recognises compact folding intermediates exposing extensive non-native surface and dictates DnaJ access to nascent chains. DnaJ uses a large surface to bind structurally diverse intermediates, and recruits DnaK to sequence-diverse solvent-accessible sites. Neither Trigger factor, DnaJ nor DnaK destabilize cotranslational folding intermediates. Instead, the chaperones collaborate to create a protected space for protein maturation that extends well beyond the ribosome exit tunnel. Our findings show how the chaperone network selects and modulates cotranslational folding intermediates.
]]></description>
<dc:creator>Roeselova, A.</dc:creator>
<dc:creator>Maslen, S. L.</dc:creator>
<dc:creator>Shivakumaraswamy, S.</dc:creator>
<dc:creator>Pellowe, G. A.</dc:creator>
<dc:creator>Howell, S.</dc:creator>
<dc:creator>Joshi, D.</dc:creator>
<dc:creator>Redmond, J.</dc:creator>
<dc:creator>Kjaer, S.</dc:creator>
<dc:creator>Skehel, M.</dc:creator>
<dc:creator>Balchin, D.</dc:creator>
<dc:date>2024-01-23</dc:date>
<dc:identifier>doi:10.1101/2024.01.22.576655</dc:identifier>
<dc:title><![CDATA[Mechanism of chaperone coordination during cotranslational protein folding in bacteria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.23.576717v1?rss=1">
<title>
<![CDATA[
Smad4 is essential for epiblast scaling and morphogenesis after implantation, but nonessential prior to implantation in the mouse 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.23.576717v1?rss=1"
</link>
<description><![CDATA[
Bone Morphogenic Protein (BMP) signaling plays an essential and highly conserved role in axial patterning in embryos of many externally developing animal species. However, in mammalian embryos, which develop inside the mother, early development includes an additional stage known as preimplantation. During preimplantation, the epiblast lineage is segregated from the extraembryonic lineages that enable implantation and development in utero. Yet, the requirement for BMP signaling in mouse preimplantation is imprecisely defined. We show that, in contrast to prior reports, BMP signaling (as reported by SMAD1/5/9 phosphorylation) is not detectable until implantation, when it is detected in the primitive endoderm - an extraembryonic lineage. Moreover, preimplantation development appears normal following deletion of maternal and zygotic Smad4, an essential effector of BMP signaling. In fact, mice lacking maternal Smad4 are viable. Finally, we uncover a new requirement for zygotic Smad4 in epiblast scaling and cavitation immediately after implantation, via a mechanism involving FGFR/ERK attenuation. Altogether, our results demonstrate no role for BMP4/SMAD4 in the first lineage decisions during mouse development. Rather, multi-pathway signaling among embryonic and extraembryonic cell types drives epiblast morphogenesis post-implantation.

Summary StatementGene expression, gene deletion, and pathway visualization evidence show that Smad4-dependent signaling is first active after mouse embryo implantation, when it promotes epiblast morphogenesis non-cell autonomously.
]]></description>
<dc:creator>Kruger, R.</dc:creator>
<dc:creator>Frum, T.</dc:creator>
<dc:creator>Brumm, A. S.</dc:creator>
<dc:creator>Hickey, S. L.</dc:creator>
<dc:creator>Niakan, K.</dc:creator>
<dc:creator>Aziz, F.</dc:creator>
<dc:creator>Shammami, M.</dc:creator>
<dc:creator>Roberts, J.</dc:creator>
<dc:creator>Ralston, A.</dc:creator>
<dc:date>2024-01-24</dc:date>
<dc:identifier>doi:10.1101/2024.01.23.576717</dc:identifier>
<dc:title><![CDATA[Smad4 is essential for epiblast scaling and morphogenesis after implantation, but nonessential prior to implantation in the mouse]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.24.576991v1?rss=1">
<title>
<![CDATA[
Context-dependent effects of CDKN2A and other 9p21 gene losses during the evolution of oesophageal cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.24.576991v1?rss=1"
</link>
<description><![CDATA[
CDKN2A is a tumour suppressor located in chromosome 9p21 and frequently lost in Barretts oesophagus (BO) and oesophageal adenocarcinoma (OAC). How CDKN2A and other 9p21 gene co-deletions affect OAC evolution remains understudied. We explored the effects of 9p21 loss in OACs and cancer progressor and non-progressor BOs with matched genomic, transcriptomic, and clinical data. Despite its cancer driver role, CDKN2A loss in BO prevents OAC initiation by counter-selecting subsequent TP53 alterations. 9p21 gene co-deletions predict poor patient survival in OAC but not BO through context-dependent effects on cell cycle, oxidative phosphorylation, and interferon response. Immune quantifications using bulk transcriptome, RNAscope and high-dimensional tissue imaging showed that IFNE loss reduces immune infiltration in BO but not OAC. Mechanistically, CDKN2A loss suppresses the maintenance of squamous epithelium, contributing to a more aggressive phenotype. Our study demonstrates context-dependent roles of cancer genes during disease evolution, with consequences for cancer detection and patient management.
]]></description>
<dc:creator>Ganguli, P.</dc:creator>
<dc:creator>Colomina-Basanta, C.</dc:creator>
<dc:creator>Mendez, A.</dc:creator>
<dc:creator>Armero-Pineiro, M.</dc:creator>
<dc:creator>Zahra, A.</dc:creator>
<dc:creator>Acha-Sagrado, A.</dc:creator>
<dc:creator>Misetic, H.</dc:creator>
<dc:creator>Devonshire, G.</dc:creator>
<dc:creator>Kelly, G.</dc:creator>
<dc:creator>Freeman, A.</dc:creator>
<dc:creator>Green, M.</dc:creator>
<dc:creator>Nye, E.</dc:creator>
<dc:creator>Oesophageal Cancer Clinical and Molecular Stratification (OCCAMS) Consortium,</dc:creator>
<dc:creator>Rodriguez-Justo, M.</dc:creator>
<dc:creator>Spencer, J.</dc:creator>
<dc:creator>Fitzgerald, R. C.</dc:creator>
<dc:creator>Ciccarelli, F. D.</dc:creator>
<dc:date>2024-01-26</dc:date>
<dc:identifier>doi:10.1101/2024.01.24.576991</dc:identifier>
<dc:title><![CDATA[Context-dependent effects of CDKN2A and other 9p21 gene losses during the evolution of oesophageal cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.26.577387v1?rss=1">
<title>
<![CDATA[
The structural and mechanistic bases for the viral resistance to allosteric HIV-1 integrase inhibitor pirmitegravir 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.26.577387v1?rss=1"
</link>
<description><![CDATA[
Allosteric HIV-1 integrase (IN) inhibitors (ALLINIs) are investigational antiretroviral agents which potently impair virion maturation by inducing hyper-multimerization of IN and inhibiting its interaction with viral genomic RNA. The pyrrolopyridine-based ALLINI pirmitegravir (PIR) has recently advanced into Phase 2a clinical trials. Previous cell culture based viral breakthrough assays identified the HIV-1(Y99H/A128T IN) variant that confers substantial resistance to this inhibitor. Here, we have elucidated the unexpected mechanism of viral resistance to PIR. While both Tyr99 and Ala128 are positioned within the inhibitor binding V-shaped cavity at the IN catalytic core domain (CCD) dimer interface, the Y99H/A128T IN mutations did not substantially affect direct binding of PIR to the CCD dimer or functional oligomerization of full-length IN. Instead, the drug-resistant mutations introduced a steric hindrance at the inhibitor mediated interface between CCD and C-terminal domain (CTD) and compromised CTD binding to the CCDY99H/A128T + PIR complex. Consequently, full-length INY99H/A128T was substantially less susceptible to the PIR induced hyper-multimerization than the WT protein, and HIV-1(Y99H/A128T IN) conferred >150- fold resistance to the inhibitor compared to the WT virus. By rationally modifying PIR we have developed its analog EKC110, which readily induced hyper-multimerization of INY99H/A128T in vitro and was [~]14-fold more potent against HIV-1(Y99H/A128T IN) than the parent inhibitor. These findings suggest a path for developing improved PIR chemotypes with a higher barrier to resistance for their potential clinical use.

IMPORTANCEAntiretroviral therapies save the lives of millions of people living with HIV (PLWH). However, evolution of multi-drug-resistant viral phenotypes is a major clinical problem, and there are limited or no treatment options for heavily treatment-experienced PLWH. Allosteric HIV-1 integrase inhibitors (ALLINIs) are a novel class of antiretroviral compounds which work by a unique mechanism of binding to the non-catalytic site on the viral protein and inducing aberrant integrase multimerization. Accordingly, ALLINIs potently inhibit both wild type HIV-1 and all drug-resistant viral phenotypes that have so far emerged against currently used therapies. Pirmitegravir, a highly potent and safe investigational ALLINI, is currently advancing through clinical trials. Here we have elucidated structural and mechanistic bases behind the emergence of HIV-1 integrase mutations in infected cell that confer resistance to pirmitegravir. In turn, our findings allowed us to rationally develop an improved ALLINI with substantially enhanced potency against the pirmitegravir resistant virus.
]]></description>
<dc:creator>Dinh, T.</dc:creator>
<dc:creator>Tber, Z.</dc:creator>
<dc:creator>Rey, J. S.</dc:creator>
<dc:creator>Mengshetti, S.</dc:creator>
<dc:creator>Annamalai, A. S.</dc:creator>
<dc:creator>Haney, R.</dc:creator>
<dc:creator>Briganti, L.</dc:creator>
<dc:creator>Amblard, F.</dc:creator>
<dc:creator>Fuchs, J. R.</dc:creator>
<dc:creator>Cherepanov, P.</dc:creator>
<dc:creator>Kim, K.</dc:creator>
<dc:creator>Schinazi, R. F.</dc:creator>
<dc:creator>Perilla, J. R.</dc:creator>
<dc:creator>Kim, B.</dc:creator>
<dc:creator>Kvaratskhelia, M.</dc:creator>
<dc:date>2024-01-26</dc:date>
<dc:identifier>doi:10.1101/2024.01.26.577387</dc:identifier>
<dc:title><![CDATA[The structural and mechanistic bases for the viral resistance to allosteric HIV-1 integrase inhibitor pirmitegravir]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.01.578443v1?rss=1">
<title>
<![CDATA[
Force-transducing molecular ensembles at growing microtubule tips control mitotic spindle size 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.01.578443v1?rss=1"
</link>
<description><![CDATA[
Mitotic spindle is a complex bipolar cellular structure that ensures chromosomes segregation between dividing cells. Correct spindle size is required for the accurate segregation and successful passing of genomes to the newly formed cells. The spindle size is believed to be controlled by mechanical forces generated by molecular motors and non-motor proteins acting in the spindle microtubule overlaps. However, how forces generated by individual proteins enable bipolar spindle organization is not well understood. Here, we developed tools to measure contributions of individual molecules to this force balance. We show that microtubule tip-trackers act synergetically at microtubule tips with minus-end directed motors to produce a system that can generate both pushing and pulling forces. We show that this system harnesses forces generated by growing tips of spindle microtubules and provides unique contribution to the force balance distinct from other force generators because it acts at microtubule tips rather than in microtubule overlaps. We show that this system alone can establish stable bipolar organization in vitro and in mitotic spindles in human cells. Our results pave the way for understanding how mechanical forces in spindles can be fine-tuned to control the fidelity of chromosome segregation.
]]></description>
<dc:creator>Chu, L.-Y.</dc:creator>
<dc:creator>Stedman, D.</dc:creator>
<dc:creator>Gannon, J.</dc:creator>
<dc:creator>Cox, S.</dc:creator>
<dc:creator>Pobegalov, G.</dc:creator>
<dc:creator>Molodtsov, M.</dc:creator>
<dc:date>2024-02-02</dc:date>
<dc:identifier>doi:10.1101/2024.02.01.578443</dc:identifier>
<dc:title><![CDATA[Force-transducing molecular ensembles at growing microtubule tips control mitotic spindle size]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.04.578801v1?rss=1">
<title>
<![CDATA[
Activation loop phosphorylation and cGMP saturation of PKG regulate egress of malaria parasites. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.04.578801v1?rss=1"
</link>
<description><![CDATA[
The cGMP-dependent protein kinase (PKG) is the sole cGMP sensor in malaria parasites, acting as an essential signalling hub to govern key developmental processes throughout the parasite life cycle. Despite the importance of PKG in the clinically relevant asexual blood stages, many aspects of malarial PKG regulation, including the importance of phosphorylation, remain poorly understood. Here we use genetic and biochemical approaches to show that reduced cGMP binding to cyclic nucleotide binding domain B does not affect in vitro kinase activity but prevents parasite egress. Similarly, we show that phosphorylation of a key threonine residue (T695) in the activation loop is dispensable for kinase activity in vitro but is essential for in vivo PKG function, with loss of T695 phosphorylation leading to aberrant phosphorylation events across the parasite proteome and changes to the substrate specificity of PKG. Our findings indicate that Plasmodium PKG is uniquely regulated to transduce signals crucial for malaria parasite development.
]]></description>
<dc:creator>Koussis, K.</dc:creator>
<dc:creator>Haase, S.</dc:creator>
<dc:creator>Withers-Martinez, C.</dc:creator>
<dc:creator>Flynn, H. R.</dc:creator>
<dc:creator>Kunzelmann, S.</dc:creator>
<dc:creator>Christodoulou, E.</dc:creator>
<dc:creator>Ibrahim, F.</dc:creator>
<dc:creator>Skehel, M.</dc:creator>
<dc:creator>Baker, D. A.</dc:creator>
<dc:creator>Blackman, M. J.</dc:creator>
<dc:date>2024-02-04</dc:date>
<dc:identifier>doi:10.1101/2024.02.04.578801</dc:identifier>
<dc:title><![CDATA[Activation loop phosphorylation and cGMP saturation of PKG regulate egress of malaria parasites.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.05.578160v1?rss=1">
<title>
<![CDATA[
Plasmodium NEK1 coordinates MTOC organisation and kinetochore attachment during rapid mitosis in male gamete formation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.05.578160v1?rss=1"
</link>
<description><![CDATA[
Mitosis is an important process in the cell cycle required for cells to divide. Never in mitosis (NIMA)-like kinases (NEKs) are regulators of mitotic functions in diverse organisms. Plasmodium spp, the causative agent of malaria is a divergent unicellular haploid eukaryote with some unusual features in terms of its mitotic and nuclear division cycle that presumably facilitate proliferation in varied environments. For example, during the sexual stage of male gametogenesis that occurs within the mosquito host, an atypical rapid closed endomitosis is observed. Three rounds of genome replication from 1N to 8N and successive cycles of multiple spindle formation and chromosome segregation occur within eight minutes followed by karyokinesis to generate haploid gametes.

Our previous Plasmodium berghei kinome screen identified four Nek genes, of which two, NEK2 and NEK4, are required for meiosis. NEK1 is likely to be essential for mitosis in asexual blood stage schizogony in the vertebrate host, but its function during male gametogenesis is unknown. Here, we study NEK1 location and function, using live cell imaging, ultrastructure expansion microscopy (U-ExM) and electron microscopy, together with conditional gene knockdown and proteomic approaches. We report spatiotemporal NEK1 location in real-time, coordinated with microtubule organising centre (MTOC) dynamics during the unusual mitoses at various stages of the Plasmodium spp. life cycle. Knockdown studies reveal NEK1 to be an essential component of the MTOC in male cell differentiation, associated with rapid mitosis, spindle formation and kinetochore attachment. These data suggest that Plasmodium berghei NEK1 kinase is an important component of MTOC organisation and essential regulator of chromosome segregation during male gamete formation.
]]></description>
<dc:creator>Zeeshan, M.</dc:creator>
<dc:creator>Rashpa, R.</dc:creator>
<dc:creator>Ferguson, D. J.</dc:creator>
<dc:creator>Mckeown, G.</dc:creator>
<dc:creator>Nugmanova, R.</dc:creator>
<dc:creator>Subodhi, A. K.</dc:creator>
<dc:creator>Beyeler, R.</dc:creator>
<dc:creator>Pashley, S. L.</dc:creator>
<dc:creator>Markus, R.</dc:creator>
<dc:creator>Brady, D.</dc:creator>
<dc:creator>Roques, M.</dc:creator>
<dc:creator>Bottrill, A. R.</dc:creator>
<dc:creator>Fry, A. M.</dc:creator>
<dc:creator>Pain, A.</dc:creator>
<dc:creator>vaughan, s.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:creator>Tromer, E. C.</dc:creator>
<dc:creator>Brochet, M.</dc:creator>
<dc:creator>Tewari, R.</dc:creator>
<dc:date>2024-02-05</dc:date>
<dc:identifier>doi:10.1101/2024.02.05.578160</dc:identifier>
<dc:title><![CDATA[Plasmodium NEK1 coordinates MTOC organisation and kinetochore attachment during rapid mitosis in male gamete formation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.05.578789v1?rss=1">
<title>
<![CDATA[
Preservation of memory B cell homeostasis in an individual producing broadly neutralising antibodies against HIV-1 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.05.578789v1?rss=1"
</link>
<description><![CDATA[
Immunological determinants favouring emergence of broadly neutralising antibodies are crucial to the development of HIV-1 vaccination strategies. Here, we combined RNAseq and B cell cloning approaches to isolate a broadly neutralising antibody (bnAb) ELC07 from an individual living with untreated HIV-1. Using single particle cryogenic electron microscopy (cryo-EM), we show that the antibody recognises a conformational epitope at the gp120-gp41 interface. ELC07 binds the closed state of the viral glycoprotein causing considerable perturbations to the gp41 trimer core structure. Phenotypic analysis of memory B cell subsets from the ELC07 bnAb donor revealed a lack of expected HIV-1-associated dysfunction, specifically no increase in CD21-/CD27- cells was observed whilst the resting memory (CD21+/CD27+) population appeared preserved despite uncontrolled HIV-1 viraemia. Moreover, single cell transcriptomes of memory B cells from this bnAb donor showed a resting memory phenotype irrespective of the epitope they targeted or their ability to neutralise diverse strains of HIV-1. Strikingly, single memory B cells from the ELC07 bnAb donor were transcriptionally similar to memory B cells from HIV-negative individuals. Our results demonstrate that potent bnAbs can arise without the HIV-1-induced dysregulation of the memory B cell compartment and suggest that sufficient levels of antigenic stimulation with a strategically designed immunogen could be effective in HIV-negative vaccine recipients.
]]></description>
<dc:creator>Griffith, S.</dc:creator>
<dc:creator>Muir, L.</dc:creator>
<dc:creator>Suchanek, O.</dc:creator>
<dc:creator>Hope, J.</dc:creator>
<dc:creator>Pade, C.</dc:creator>
<dc:creator>Gibbons, J. M.</dc:creator>
<dc:creator>Tuong, Z. K.</dc:creator>
<dc:creator>Fung, A.</dc:creator>
<dc:creator>Touizer, E.</dc:creator>
<dc:creator>Rees-Spear, C.</dc:creator>
<dc:creator>Nans, A.</dc:creator>
<dc:creator>Roustan, C.</dc:creator>
<dc:creator>Alguel, Y.</dc:creator>
<dc:creator>Fink, D.</dc:creator>
<dc:creator>Orkin, C.</dc:creator>
<dc:creator>Deayton, J.</dc:creator>
<dc:creator>Anderson, J.</dc:creator>
<dc:creator>Gupta, R. K.</dc:creator>
<dc:creator>Doores, K. J.</dc:creator>
<dc:creator>Cherepanov, P.</dc:creator>
<dc:creator>McKnight, A.</dc:creator>
<dc:creator>Clatworthy, M.</dc:creator>
<dc:creator>McCoy, L. E.</dc:creator>
<dc:date>2024-02-06</dc:date>
<dc:identifier>doi:10.1101/2024.02.05.578789</dc:identifier>
<dc:title><![CDATA[Preservation of memory B cell homeostasis in an individual producing broadly neutralising antibodies against HIV-1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.06.579027v1?rss=1">
<title>
<![CDATA[
Focal Deletions of a Promoter Tether Activate the IRX3 Oncogene in T Cell Acute Lymphoblastic Leukemia 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.06.579027v1?rss=1"
</link>
<description><![CDATA[
Oncogenes can be activated in cis through multiple mechanisms including enhancer hijacking events and noncoding mutations that create enhancers or promoters de novo. These paradigms have helped parse somatic variation of noncoding cancer genomes, thereby providing a rationale to identify noncanonical mechanisms of gene activation. Here we describe a novel mechanism of oncogene activation whereby focal copy number loss of an intronic element within the FTO gene leads to aberrant expression of IRX3, an oncogene in T cell acute lymphoblastic leukemia (T-ALL). Loss of this CTCF bound element downstream to IRX3 (+224 kb) leads to enhancer hijack of an upstream developmentally active super-enhancer of the CRNDE long noncoding RNA (-644 kb). Unexpectedly, the CRNDE super-enhancer interacts with the IRX3 promoter with no transcriptional output until it is untethered from the FTO intronic site. We propose that  promoter tethering of oncogenes to inert regions of the genome is a previously unappreciated biological mechanism preventing tumorigenesis.
]]></description>
<dc:creator>Rahman, S.</dc:creator>
<dc:creator>Bloye, G.</dc:creator>
<dc:creator>Farah, N.</dc:creator>
<dc:creator>Demeulemeester, J.</dc:creator>
<dc:creator>Costa, J. R.</dc:creator>
<dc:creator>O'Connor, D.</dc:creator>
<dc:creator>Pocock, R.</dc:creator>
<dc:creator>Turna, A.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Fielding, A. K.</dc:creator>
<dc:creator>Roels, J.</dc:creator>
<dc:creator>Jaksik, R.</dc:creator>
<dc:creator>Dawidowska, M.</dc:creator>
<dc:creator>Vlierberghe, P. V.</dc:creator>
<dc:creator>Hadjur, S.</dc:creator>
<dc:creator>Hughes, J. R.</dc:creator>
<dc:creator>Davies, J. O. J.</dc:creator>
<dc:creator>Gutierrez, A.</dc:creator>
<dc:creator>Kelliher, M. A.</dc:creator>
<dc:creator>Loo, P. V.</dc:creator>
<dc:creator>Dawson, M. A.</dc:creator>
<dc:creator>Mansour, M. R.</dc:creator>
<dc:date>2024-02-08</dc:date>
<dc:identifier>doi:10.1101/2024.02.06.579027</dc:identifier>
<dc:title><![CDATA[Focal Deletions of a Promoter Tether Activate the IRX3 Oncogene in T Cell Acute Lymphoblastic Leukemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.10.579757v1?rss=1">
<title>
<![CDATA[
Zebrafish reveal new roles for Fam83f in hatching and the DNA damage-mediated autophagic response 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.10.579757v1?rss=1"
</link>
<description><![CDATA[
The FAM83 (Family with sequence similarity 83) family is highly conserved in vertebrates, but little is known of the functions of these proteins beyond their association with oncogenesis. Of the family, FAM83F is of particular interest because it is the only membrane-targeted FAM83 protein. When over-expressed, FAM83F activates the canonical Wnt signalling pathway and binds to and stabilizes p53; it therefore interacts with two pathways often dysregulated in disease. Insights into gene function can often be gained by studying the roles they play during development, and here we report the generation of fam83f knock-out (KO) zebrafish, which we have used to study the role of Fam83f in vivo. We show that endogenous fam83f is most strongly expressed in the hatching gland of developing zebrafish embryos, and that fam83f KO embryos hatch earlier than their wild-type (WT) counterparts, despite developing at a comparable rate. We also demonstrate that fam83f KO embryos are more sensitive to ionizing radiation than WT embryos--an unexpected finding, bearing in mind the previously-reported ability of FAM83F to stabilize p53. Transcriptomic analysis shows that loss of fam83f leads to downregulation of phosphatidylinositol-3-phosphate (PI(3)P) binding proteins and impairment of cellular degradation pathways, particularly autophagy, a crucial component of the DNA damage response. Finally, we show that Fam83f protein is itself targeted to the lysosome when over-expressed in HEK293T cells, and that this localization is dependent upon a C terminal signal sequence. The zebrafish lines we have generated suggest that Fam83f plays an important role in autophagic/lysosomal processes, resulting in dysregulated hatching and increased sensitivity to genotoxic stress in vivo.
]]></description>
<dc:creator>Jones, R. A.</dc:creator>
<dc:creator>Cooper, F.</dc:creator>
<dc:creator>Kelly, G.</dc:creator>
<dc:creator>Barry, D.</dc:creator>
<dc:creator>Renshaw, M. J.</dc:creator>
<dc:creator>Sapkota, G.</dc:creator>
<dc:creator>Smith, J. C.</dc:creator>
<dc:date>2024-02-12</dc:date>
<dc:identifier>doi:10.1101/2024.02.10.579757</dc:identifier>
<dc:title><![CDATA[Zebrafish reveal new roles for Fam83f in hatching and the DNA damage-mediated autophagic response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.01.578414v1?rss=1">
<title>
<![CDATA[
Suppression of ERK signalling promotes pluripotent epiblast in the human blastocyst 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.01.578414v1?rss=1"
</link>
<description><![CDATA[
Studies in the mouse demonstrate the importance of fibroblast growth factor (FGF) and extra-cellular receptor tyrosine kinase (ERK) in specification of embryo-fated epiblast and yolk-sac-fated hypoblast cells from uncommitted inner cell mass (ICM) cells prior to implantation. Molecular mechanisms regulating specification of early lineages in human development are comparatively unclear. Here we show that exogenous FGF stimulation leads to expanded hypoblast molecular marker expression, at the expense of the epiblast. Conversely, we show that specifically inhibiting ERK activity leads to expansion of epiblast cells functionally capable of giving rise to naive human pluripotent stem cells. Single-cell transcriptomic analysis indicates that these epiblast cells downregulate FGF signalling and upregulate molecular markers associated with naive pluripotency. Our functional study demonstrates for the first time the molecular mechanisms governing ICM specification in human development, whereby segregation of the epiblast and hypoblast lineages occurs during maturation of the mammalian embryo in an ERK signal-dependent manner.
]]></description>
<dc:creator>Simon, C. S.</dc:creator>
<dc:creator>McCarthy, A.</dc:creator>
<dc:creator>Woods, L.</dc:creator>
<dc:creator>Staneva, D.</dc:creator>
<dc:creator>Huang, Q.</dc:creator>
<dc:creator>Linneberg-Agerholm, M.</dc:creator>
<dc:creator>Faulkner, A.</dc:creator>
<dc:creator>Papathanasiou, A.</dc:creator>
<dc:creator>Elder, K.</dc:creator>
<dc:creator>Snell, P.</dc:creator>
<dc:creator>Christie, L.</dc:creator>
<dc:creator>Garcia, P.</dc:creator>
<dc:creator>Shaikly, V.</dc:creator>
<dc:creator>Taranissi, M.</dc:creator>
<dc:creator>Choudhary, M.</dc:creator>
<dc:creator>Herbert, M.</dc:creator>
<dc:creator>Brickman, J. M.</dc:creator>
<dc:creator>Niakan, K. K.</dc:creator>
<dc:date>2024-02-04</dc:date>
<dc:identifier>doi:10.1101/2024.02.01.578414</dc:identifier>
<dc:title><![CDATA[Suppression of ERK signalling promotes pluripotent epiblast in the human blastocyst]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.12.579991v1?rss=1">
<title>
<![CDATA[
Micromolar fluoride contamination arising from glass NMR tubes and a simple solution for biomolecular applications 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.12.579991v1?rss=1"
</link>
<description><![CDATA[
Fluorine (19F) NMR is emerging as an invaluable analytical technique in chemistry, biochemistry, material science, and medicine, especially due to the inherent rarity of naturally occurring fluorine in biological, organic, and inorganic compounds. Thus, we were surprised to identify an unexpected peak in our 19F NMR spectra, corresponding to free fluoride, which appears to leach out from various types of new and unused glass NMR tubes over the course of several hours. We quantified this contaminant to be at micromolar concentrations for typical NMR sample volumes across multiple glass types and brands. We find that this artefact is undetectable for samples prepared in quartz NMR tubes within the timeframes of our experiments. We also observed that pre-soaking new glass NMR tubes combined with rinsing removes this contamination below micromolar levels. Given the increasing popularity of 19F NMR across a wide range of fields, the long collection times required for relaxation studies and samples of low concentrations, and the importance of avoiding contamination in all NMR experiments, we anticipate that our simple solution will be useful to biomolecular NMR spectroscopists.
]]></description>
<dc:creator>Matwani, K.</dc:creator>
<dc:creator>Cornish, J.</dc:creator>
<dc:creator>DeBenedictis, E. A.</dc:creator>
<dc:creator>Heller, G. T.</dc:creator>
<dc:date>2024-02-12</dc:date>
<dc:identifier>doi:10.1101/2024.02.12.579991</dc:identifier>
<dc:title><![CDATA[Micromolar fluoride contamination arising from glass NMR tubes and a simple solution for biomolecular applications]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.13.580055v1?rss=1">
<title>
<![CDATA[
Optical tweezers reveal that PfEBA and PfRH ligands, not PfMSP1, play a central role in Plasmodium-falciparum merozoite-erythrocyte attachment 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.13.580055v1?rss=1"
</link>
<description><![CDATA[
Malaria pathogenesis and parasite multiplication both depend on the ability of Plasmodium falciparum merozoites to invade human erythrocytes. Invasion is a complex multi-step process that is known to involve multiple P. falciparum proteins but dissecting the precise role of individual proteins has to date been limited by the availability of quantifiable phenotypic assays. In this study, we apply a new approach to assigning function to invasion proteins by using optical tweezers to directly manipulate recently egressed merozoites and erythrocytes and quantify the strength of attachment between them, as well as the frequency with which such attachments occur. Using a range of inhibitors, antibodies, and genetically modified P. falciparum strains, we quantitated the contribution of individual P. falciparum proteins to these merozoite-erythrocyte attachment phenotypes for the first time. Most of the interactions investigated did not affect the force needed to pull merozoites and erythrocytes apart, including loss of the major P. falciparum merozoite surface protein PfMSP1 and PfGAP45, part of the glideosome actinomyosin motor complex. The only factors that significantly reduced the strength of merozoite-erythrocyte attachment were ones that disrupted the function of members of the EBA-175 like Antigen (PfEBA) family and Reticulocyte Binding Protein Homologue (PfRH) invasion ligand families. While these assays also reinforced the known redundancy within these families, with the deletion of some ligands not impacting detachment force, it appears that the PfEBA/PfRH families play a central role in merozoite attachment, not the major merozoite surface protein PfMSP1.

Author summaryMalaria is a devastating disease caused by a parasitic infection. The deadliest species is Plasmodium falciparum, which causes more than 600,000 deaths annually. The parasites life cycle is complex, but all the symptoms of malaria are caused when the parasites replicate in human red blood cells. Replication depends on the invasion of the red blood cells by the parasites which is a complex process involving multiple molecular interactions and multiple steps. Invasion begins with the attachment of the parasite to the red blood cell, making this step of particular interest in the development of new therapeutics. We assessed which interactions are key to the strength of attachment using an optical tweezer assay, which allowed us to directly measure the binding force between individual parasites and red blood cells whilst using a range of molecular and genetic tools that target specific interactions known to have a role in invasion. This showed that loss of a protein commonly thought to be critical to the early stages of binding (PfMSP1) had no effect on attachment strength, whereas disruptions of several members from two families of proteins (the Erythrocyte Binding Like protein family and the reticulocyte binding-like protein family) affect attachment strength.
]]></description>
<dc:creator>Kals, E.</dc:creator>
<dc:creator>Kals, M.</dc:creator>
<dc:creator>Lees, R. A.</dc:creator>
<dc:creator>Introini, V.</dc:creator>
<dc:creator>Kemp, A.</dc:creator>
<dc:creator>Silvester, E.</dc:creator>
<dc:creator>Collins, C. R.</dc:creator>
<dc:creator>Umrekar, T.</dc:creator>
<dc:creator>Kotar, J.</dc:creator>
<dc:creator>Cicuta, P.</dc:creator>
<dc:creator>Rayner, J. C.</dc:creator>
<dc:date>2024-02-13</dc:date>
<dc:identifier>doi:10.1101/2024.02.13.580055</dc:identifier>
<dc:title><![CDATA[Optical tweezers reveal that PfEBA and PfRH ligands, not PfMSP1, play a central role in Plasmodium-falciparum merozoite-erythrocyte attachment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.17.580698v1?rss=1">
<title>
<![CDATA[
Large-scale visualisation of α-synuclein oligomers in Parkinson's disease brain tissue 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.17.580698v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is a common neurodegenerative condition characterised by the presence in the brain of large intraneuronal aggregates, known as Lewy bodies and Lewy neurites, containing fibrillar -synuclein. According to the amyloid hypothesis, these large end-stage species form from smaller soluble protein assemblies, often termed oligomers, which are proposed as early drivers of pathogenesis. To date, however, this hypothesis has remained controversial, at least in part because it has not been possible to directly visualise oligomeric aggregates in human brain tissue. Therefore, their presence, abundance and distributions have remained elusive. Here, we present ASA-PD (Advanced Sensing of Aggregates - Parkinsons Disease), an imaging method to generate large-scale -synuclein oligomer maps in post-mortem human brain tissue. We combined autofluorescence suppression with single-molecule fluorescence methods, which together, enable the detection of nanoscale -synuclein aggregates. To demonstrate the utility of this platform, we captured [~]1.2 million oligomers from the anterior cingulate cortex in human post-mortem brain samples from PD and healthy control patients. Our data revealed a specific subpopulation of nanoscale oligomers that represent an early hallmark of the proteinopathy that underlies PD. We anticipate that quantitative information about oligomer distributions provided by ASA-PD will enable mechanistic studies to reveal the pathological processes caused by -synuclein aggregation.
]]></description>
<dc:creator>Andrews, R.</dc:creator>
<dc:creator>Fu, B.</dc:creator>
<dc:creator>Toomey, C. E.</dc:creator>
<dc:creator>Breiter, J. C.</dc:creator>
<dc:creator>Lachica, J.</dc:creator>
<dc:creator>Tian, R.</dc:creator>
<dc:creator>Beckwith, J. S.</dc:creator>
<dc:creator>Needham, L.-M.</dc:creator>
<dc:creator>Chant, G. J.</dc:creator>
<dc:creator>Loiseau, C.</dc:creator>
<dc:creator>Deconfin, A.</dc:creator>
<dc:creator>Baspin, K.</dc:creator>
<dc:creator>Magill, P. J.</dc:creator>
<dc:creator>Jaunmuktane, Z.</dc:creator>
<dc:creator>Freeman, O. J.</dc:creator>
<dc:creator>Taylor, B. J. M.</dc:creator>
<dc:creator>Hardy, J.</dc:creator>
<dc:creator>Lashley, T.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:creator>Vendruscolo, M.</dc:creator>
<dc:creator>Wood, N. W.</dc:creator>
<dc:creator>Weiss, L. E.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Lee, S. F.</dc:creator>
<dc:date>2024-02-19</dc:date>
<dc:identifier>doi:10.1101/2024.02.17.580698</dc:identifier>
<dc:title><![CDATA[Large-scale visualisation of α-synuclein oligomers in Parkinson's disease brain tissue]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.14.580367v1?rss=1">
<title>
<![CDATA[
A single amino acid in the Salmonella effector SarA/SteE triggers supraphysiological activation of STAT3 for anti-inflammatory target gene expression 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.14.580367v1?rss=1"
</link>
<description><![CDATA[
Non-typhoidal Salmonella enterica cause an estimated 1 million cases of gastroenteritis annually in the United States. These serovars use secreted protein effectors to mimic and reprogram host cellular functions. We previously discovered that the secreted effector SarA (Salmonella anti-inflammatory response activator; also known as SteE) was required for increased intracellular replication of S. Typhimurium and production of the anti-inflammatory cytokine interleukin-10 (IL-10). SarA facilitates phosphorylation of STAT3 through a region of homology with the host cytokine receptor gp130. Here, we demonstrate that a single amino acid difference between SarA and gp130 is critical for the anti-inflammatory bias of SarA-STAT3 signaling. An isoleucine at the pY+1 position of the YxxQ motif in SarA (which binds the SH2 domain in STAT3) causes increased STAT3 phosphorylation and expression of anti-inflammatory target genes. This isoleucine, completely conserved in [~]4000 Salmonella isolates, renders SarA a better substrate for tyrosine phosphorylation by GSK-3. GSK-3 is canonically a serine/threonine kinase that nonetheless undergoes tyrosine autophosphorylation at a motif that has an invariant isoleucine at the pY+1 position. Our results provide a molecular basis for how a Salmonella secreted effector achieves supraphysiological levels of STAT3 activation to control host genes during infection.
]]></description>
<dc:creator>Gaggioli, M. R.</dc:creator>
<dc:creator>Jones, A. G.</dc:creator>
<dc:creator>Panagi, I.</dc:creator>
<dc:creator>Washington, E. J.</dc:creator>
<dc:creator>Loney, R. E.</dc:creator>
<dc:creator>Muench, J. H.</dc:creator>
<dc:creator>Brennan, R. G.</dc:creator>
<dc:creator>Thurston, T. L. M.</dc:creator>
<dc:creator>Ko, D. C.</dc:creator>
<dc:date>2024-02-14</dc:date>
<dc:identifier>doi:10.1101/2024.02.14.580367</dc:identifier>
<dc:title><![CDATA[A single amino acid in the Salmonella effector SarA/SteE triggers supraphysiological activation of STAT3 for anti-inflammatory target gene expression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.22.581535v1?rss=1">
<title>
<![CDATA[
Evolution and inhibition of the FIKK effector kinase family in P. falciparum 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.22.581535v1?rss=1"
</link>
<description><![CDATA[
Among the [~]200 Plasmodium species that infect vertebrates, six infect humans. Of these, P. falciparum causes >95% of all [~]500,000 annual fatalities. Phylogenetically, P. falciparum belongs to the Laverania subgenus, a group of Plasmodium species that infect great apes. Common to Laverania species is the family of FIKK kinases. One million years ago, a single FIKK kinase conserved in all Plasmodium species gained an export element in the Laverania subgenus and expanded into the family of [~]20 atypical FIKK kinases, most of which are exported into the host cell. The fikk genes are conserved in syntenic loci across the Laverania, arguing for a rapid expansion controlling important functions in host cell remodelling and pathogenesis. We provide evidence that the FIKK paralogues evolved specific and mutually exclusive phosphorylation motif preferences, conserved across their Laverania orthologues, in a short evolutionary timeframe. Surprisingly, we find that FIKK13 has evolved exclusive tyrosine-phosphorylation preference, which was thought to be absent in Plasmodium species. Combining a crystal structure with AlphaFold2 predictions, we identify residues that determine kinase-specificity within the FIKK family in a fast-evolving flexible loop. Finally, we show that all expressed members of the FIKK kinase family can be chemically inhibited in vitro using a single compound. Such a pan-specific inhibitor of this kinase family important for virulence could reduce the ability of the parasite to gain escape-mutations and resistance.
]]></description>
<dc:creator>Belda, H.</dc:creator>
<dc:creator>Bradley, D.</dc:creator>
<dc:creator>Christodoulou, E.</dc:creator>
<dc:creator>Nofal, S. D.</dc:creator>
<dc:creator>Broncel, M.</dc:creator>
<dc:creator>Jones, D. A.</dc:creator>
<dc:creator>Davies, H.</dc:creator>
<dc:creator>Bertran, M. T.</dc:creator>
<dc:creator>Purkiss, A. G.</dc:creator>
<dc:creator>Ogrodowicz, R. W.</dc:creator>
<dc:creator>Joshi, D.</dc:creator>
<dc:creator>O'Reilly, N.</dc:creator>
<dc:creator>Walport, L.</dc:creator>
<dc:creator>Claessens, A.</dc:creator>
<dc:creator>Powell, A.</dc:creator>
<dc:creator>House, D.</dc:creator>
<dc:creator>Kjaer, S.</dc:creator>
<dc:creator>Landry, C. R.</dc:creator>
<dc:creator>Treeck, M.</dc:creator>
<dc:date>2024-02-23</dc:date>
<dc:identifier>doi:10.1101/2024.02.22.581535</dc:identifier>
<dc:title><![CDATA[Evolution and inhibition of the FIKK effector kinase family in P. falciparum]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.26.580534v1?rss=1">
<title>
<![CDATA[
A targetable PREX2/RAC1/PI3Kβ signalling axis confers resistance to clinically relevant therapeutic approaches in melanoma 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.26.580534v1?rss=1"
</link>
<description><![CDATA[
Metastatic melanoma remains a major clinical challenge. Large-scale genomic sequencing of melanoma has identified bona fide activating mutations in RAC1, with mutations of its upstream regulator, the RAC-GEF PREX2, also commonly detected. Crucially, RAC1 mutations are associated with resistance to BRAF-targeting therapies. Despite the role of its homologue PREX1 in melanomagenesis, and evidence that some truncating PREX2 mutations drive increased RAC1 activity, no hotspot mutations have been identified, and the impact of PREX2 mutation remains contentious. Here, we use genetically engineered mouse models and patient-derived BRAFV600E-driven melanoma cell lines to dissect the role of PREX2 in melanomagenesis and response to therapy. We show that while PREX2 is dispensable for the initiation and progression of melanoma, its loss confers sensitivity to clinically relevant therapeutics. Importantly, genetic and pharmacological targeting of the RAC1 effector kinase PI3K{beta} phenocopies PREX2 loss, sensitizing our model systems to therapy. Our data reveal a druggable PREX2/RAC1/PI3K{beta} signalling axis in BRAF-mutant melanoma that could be exploited clinically.

Statement of SignificanceMetastatic melanoma remains both a clinical problem, and an opportunity for therapeutic benefit. Co-targeting of the MAPK pathway and the PREX2/RAC1/PI3K{beta} has remarkable efficacy and outperforms monotherapy MAPK targeting in vivo.
]]></description>
<dc:creator>Ford, C. A.</dc:creator>
<dc:creator>Koludrovic, D.</dc:creator>
<dc:creator>Centeno, P. P.</dc:creator>
<dc:creator>Foth, M.</dc:creator>
<dc:creator>Tsonou, E.</dc:creator>
<dc:creator>Vlahov, N.</dc:creator>
<dc:creator>Sphyris, N.</dc:creator>
<dc:creator>Gilroy, K.</dc:creator>
<dc:creator>Bull, C.</dc:creator>
<dc:creator>Nixon, C.</dc:creator>
<dc:creator>Serrels, B.</dc:creator>
<dc:creator>Munro, A. F.</dc:creator>
<dc:creator>Dawson, J. C.</dc:creator>
<dc:creator>Carragher, N. O.</dc:creator>
<dc:creator>Pavet, V.</dc:creator>
<dc:creator>Hornigold, D. C.</dc:creator>
<dc:creator>Dunne, P. D.</dc:creator>
<dc:creator>Downward, J.</dc:creator>
<dc:creator>Welch, H. C. E.</dc:creator>
<dc:creator>Barry, S. T.</dc:creator>
<dc:creator>Sansom, O. J.</dc:creator>
<dc:creator>Campbell, A. D.</dc:creator>
<dc:date>2024-02-28</dc:date>
<dc:identifier>doi:10.1101/2024.02.26.580534</dc:identifier>
<dc:title><![CDATA[A targetable PREX2/RAC1/PI3Kβ signalling axis confers resistance to clinically relevant therapeutic approaches in melanoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.26.582055v1?rss=1">
<title>
<![CDATA[
Alpha-synuclein aggregates trigger anti-viral immune pathways and RNA editing in human astrocytes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.26.582055v1?rss=1"
</link>
<description><![CDATA[
RNA editing is a post transcriptional mechanism that targets changes in RNA transcripts to modulate innate immune responses. We report the role of astrocyte specific, ADAR1 mediated RNA editing in neuroinflammation in Parkinsons disease. We generated hiPSC-derived astrocytes, neurons and co-cultures and exposed them to small soluble alpha-synuclein aggregates. Oligomeric alpha-synuclein triggered an inflammatory glial state associated with TLR activation, viral responses, and cytokine secretion. This reactive state resulted in loss of neurosupportive functions, and the induction of neuronal toxicity. Notably, interferon response pathways were activated leading to upregulation, and isoform switching of the RNA deaminase enzyme, ADAR1. ADAR1 mediates A-to-I RNA editing, and increases in RNA editing were observed in inflammatory pathways in cells, as well as in post-mortem human PD brain. Aberrant, or dysregulated, ADAR1 responses and RNA editing may lead to sustained inflammatory reactive states in astrocytes triggered by alpha-synuclein aggregation, and this may drive the neuroinflammatory cascade in Parkinsons.
]]></description>
<dc:creator>D'Sa, K.</dc:creator>
<dc:creator>Choi, M. L.</dc:creator>
<dc:creator>Wagen, A. Z.</dc:creator>
<dc:creator>Seto-Salvia, N.</dc:creator>
<dc:creator>Kopach, O.</dc:creator>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>Rodrigues, M.</dc:creator>
<dc:creator>Lopez-Garcia, P.</dc:creator>
<dc:creator>Ghareeb, A.</dc:creator>
<dc:creator>Bayne, J.</dc:creator>
<dc:creator>Grant-Peters, M.</dc:creator>
<dc:creator>Garcia-Ruiz, S.</dc:creator>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Rodriques, S.</dc:creator>
<dc:creator>Athauda, D.</dc:creator>
<dc:creator>Gustavsson, E.</dc:creator>
<dc:creator>Gagliano Taliun, S. A.</dc:creator>
<dc:creator>Reynolds, R. H.</dc:creator>
<dc:creator>Young, G.</dc:creator>
<dc:creator>Strohbuecker, S.</dc:creator>
<dc:creator>Warner, T.</dc:creator>
<dc:creator>Rusakov, D. A.</dc:creator>
<dc:creator>Patani, R.</dc:creator>
<dc:creator>Bryant, C.</dc:creator>
<dc:creator>Klenerman, D. A.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:date>2024-02-28</dc:date>
<dc:identifier>doi:10.1101/2024.02.26.582055</dc:identifier>
<dc:title><![CDATA[Alpha-synuclein aggregates trigger anti-viral immune pathways and RNA editing in human astrocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.28.582460v1?rss=1">
<title>
<![CDATA[
GLP-1 receptor agonism ameliorates Parkinsons disease through modulation of neuronal insulin signalling and glial suppression 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.28.582460v1?rss=1"
</link>
<description><![CDATA[
Neuronal insulin resistance is linked to the pathogenesis of Parkinsons disease through unclear, but potentially targetable, mechanisms. We delineated neuronal and glial mechanisms of insulin resistance and glucagon-like 1 peptide (GLP-1) receptor agonism in human iPSC models of synucleinopathy, and corroborated our findings in patient samples from a Phase 2 trial of a GLP-1R agonist in Parkinsons (NCT01971242). Human iPSC models of synucleinopathy exhibit neuronal insulin resistance and dysfunctional insulin signalling, which is associated with inhibition of the neuroprotective Akt pathways, and increased expression of the MAPK-associated p38 and JNK stress pathways. Ultimately, this imbalance is associated with cellular stress, impaired proteostasis, accumulation of -synuclein, and neuronal loss. The GLP-1R agonist exenatide led to restoration of insulin signalling, associated with restoration of Akt signalling and suppression of the MAPK pathways in neurons. GLP-1R agonism reverses the neuronal toxicity associated with the synucleinopathy, through reduction of oxidative stress, improved mitochondrial and lysosomal function, reduced aggregation of -synuclein, and enhanced neuronal viability. GLP-1R agonism further suppresses synuclein induced inflammatory states in glia, leading to neuroprotection through non cell autonomous effects. In the exenatide-PD2 clinical trial, exenatide treatment was associated with clinical improvement in individuals with higher baseline MAPK expression (and thus insulin resistance). Exenatide treatment led to a reduction of -synuclein aggregates, and a reduction in inflammatory cytokine IL-6. Taken together, our patient platform defines the mechanisms of GLP-1R action in neurons and astrocytes, identifies the population likely to benefit from GLP-1R agonism, and highlights the utility of GLP-1R agonism as a disease modifying strategy in synucleinopathies.
]]></description>
<dc:creator>Athauda, D.</dc:creator>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>Sanchez, L.</dc:creator>
<dc:creator>Virdi, G.</dc:creator>
<dc:creator>Lopez Garcia, P.</dc:creator>
<dc:creator>Wernick, A.</dc:creator>
<dc:creator>Wagen, A.</dc:creator>
<dc:creator>D'Sa, K.</dc:creator>
<dc:creator>Lachica, J.</dc:creator>
<dc:creator>Strohbuecker, S.</dc:creator>
<dc:creator>Vecchi, G.</dc:creator>
<dc:creator>Leighton, C.</dc:creator>
<dc:creator>Saleeb, R. S.</dc:creator>
<dc:creator>OShaughnessy, J.</dc:creator>
<dc:creator>Toomey, C.</dc:creator>
<dc:creator>Vijiaratnam, N.</dc:creator>
<dc:creator>Girges, C.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Mustapic, M.</dc:creator>
<dc:creator>Ismail, K.</dc:creator>
<dc:creator>Davies, M.</dc:creator>
<dc:creator>Kapogiannis, D.</dc:creator>
<dc:creator>Choi, M. L.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:creator>Horrocks, M. H.</dc:creator>
<dc:creator>Greig, N. H.</dc:creator>
<dc:creator>Foltynie, T.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:date>2024-02-28</dc:date>
<dc:identifier>doi:10.1101/2024.02.28.582460</dc:identifier>
<dc:title><![CDATA[GLP-1 receptor agonism ameliorates Parkinsons disease through modulation of neuronal insulin signalling and glial suppression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.26.582231v1?rss=1">
<title>
<![CDATA[
TrajectoryGeometry suggests cell fate decisions involve branches rather than bifurcations 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.26.582231v1?rss=1"
</link>
<description><![CDATA[
AO_SCPLOWBSTRACTC_SCPLOWDifferentiation of multipotential progenitor cells is a key process in the development of any multi-cellular organism and often continues throughout its life. It is often assumed that a bi-potential progenitor develops along a (relatively) straight trajectory until it reaches a decision point where the trajectory bifurcates. At this point one of two directions is chosen, each direction representing the unfolding of a new transcriptomic programme. However, we have lacked quantitative means for testing this model. Accordingly, we have developed the R package TrajectoryGeometry. Applying this to published data we find several examples where, rather than bifurcate, developmental pathways branch. That is, the bipotential progenitor develops along a relatively straight trajectory leading to one of its potential fates. A second relatively straight trajectory branches off from this towards the other potential fate. In this sense only cells that branch off to follow the second trajectory make a "decision". Our methods give precise descriptions of the genes and cellular pathways involved in these trajectories. We speculate that branching may be the more common behaviour and may have advantages from a control-theoretic viewpoint.
]]></description>
<dc:creator>Laddach, A. C.</dc:creator>
<dc:creator>Shapiro, M.</dc:creator>
<dc:date>2024-02-28</dc:date>
<dc:identifier>doi:10.1101/2024.02.26.582231</dc:identifier>
<dc:title><![CDATA[TrajectoryGeometry suggests cell fate decisions involve branches rather than bifurcations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.29.582718v1?rss=1">
<title>
<![CDATA[
The Molecular Landscape of Cellular Metal Ion Biology 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.29.582718v1?rss=1"
</link>
<description><![CDATA[
Metal ions play crucial roles in cells, yet the broader impact of metal availability on biological networks remains underexplored. We generated genome-wide resources, systematically quantifying yeast cell growth, metallomic, proteomic, and genetic responses upon varying each of its essential metal ions (Ca, Cu, Fe, K, Mg, Mn, Mo, Na, Zn), over several orders of magnitude. We find that metal ions deeply impact cellular networks, with 57.6% of the proteome, including most signalling pathways, responding. While the biological response to each metal is distinct, our data reveals common properties of metal responsiveness, such as concentration interdependencies and metal homeostasis. We describe a compendium of metal-dependent cellular processes and reveal that several understudied genes can be functionally annotated based on their metal responses. Furthermore, we report that metalloenzymes occupy central nodes in the metabolic network and are more likely to be encoded by isozymes, resulting in system-wide responsiveness to metal availability.
]]></description>
<dc:creator>Aulakh, S. K.</dc:creator>
<dc:creator>Lemke, O.</dc:creator>
<dc:creator>Szyrwiel, L.</dc:creator>
<dc:creator>Kamrad, S.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Hartl, J.</dc:creator>
<dc:creator>Muelleder, M.</dc:creator>
<dc:creator>Nielsen, J.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:date>2024-03-03</dc:date>
<dc:identifier>doi:10.1101/2024.02.29.582718</dc:identifier>
<dc:title><![CDATA[The Molecular Landscape of Cellular Metal Ion Biology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.03.06.581755v1?rss=1">
<title>
<![CDATA[
A male-essential microRNA is key for avian sex chromosome dosage compensation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.03.06.581755v1?rss=1"
</link>
<description><![CDATA[
Birds have a sex chromosome system in which females are heterogametic (ZW) and males are homogametic (ZZ). The differentiation of avian sex chromosomes from ancestral autosomes entailed the loss of most genes from the W chromosome during evolution. However, to what extent mechanisms evolved that counterbalance the consequences of this extensive gene dosage reduction in female birds has remained unclear. Here we report functional in vivo and evolutionary analyses of a Z-chromosome-linked microRNA (miR-2954) with strongly male-biased expression that was previously proposed to play a key role in sex chromosome dosage compensation1. We knocked out miR-2954 in chicken, which resulted in early embryonic lethality of homozygous knockout males, likely due to the highly specific upregulation of dosage-sensitive Z-linked target genes of miR-2954. Our evolutionary gene expression analyses further revealed that these dosage-sensitive target genes have become upregulated on the single Z in female birds during evolution. Altogether, our work unveils a scenario where evolutionary pressures on females following W gene loss led to the evolution of transcriptional upregulation of dosage-sensitive genes on the Z not only in female but also in male birds. The resulting overabundance of transcripts in males resulting from the combined activity of two dosage-sensitive Z gene copies was in turn offset by the emergence of a highly targeted miR-2954-mediated transcript degradation mechanism during avian evolution. Our findings demonstrate that birds have evolved a unique sex chromosome dosage compensation system in which a microRNA has become essential for male survival.
]]></description>
<dc:creator>Fallahshahroudi, A.</dc:creator>
<dc:creator>Rodriguez-Montes, L.</dc:creator>
<dc:creator>Yousefi Taemeh, S.</dc:creator>
<dc:creator>Trost, N.</dc:creator>
<dc:creator>Tellez, M.</dc:creator>
<dc:creator>Ballantyne, M.</dc:creator>
<dc:creator>Idoko-Akoh, A.</dc:creator>
<dc:creator>Taylor, L.</dc:creator>
<dc:creator>Sherman, A.</dc:creator>
<dc:creator>Sorato, E.</dc:creator>
<dc:creator>Johnsson, M.</dc:creator>
<dc:creator>Cardoso Moreira, M.</dc:creator>
<dc:creator>McGrew, M.</dc:creator>
<dc:creator>Kaessmann, H.</dc:creator>
<dc:date>2024-03-07</dc:date>
<dc:identifier>doi:10.1101/2024.03.06.581755</dc:identifier>
<dc:title><![CDATA[A male-essential microRNA is key for avian sex chromosome dosage compensation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.09.24.461247v1?rss=1">
<title>
<![CDATA[
Solvent Precipitation SP3 (SP4) enhances recovery for proteomics sample preparation without magnetic beads 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.09.24.461247v1?rss=1"
</link>
<description><![CDATA[
Complete, reproducible extraction of protein material is essential for comprehensive and unbiased proteome analyses. A current gold standard is single-pot, solid-phase-enhanced sample preparation (SP3), in which organic solvent and magnetic beads are used to denature and capture proteins, with subsequently washes allowing contaminant removal. However, SP3 is dependent on effective protein immobilisation onto beads, risks losses during wash steps, and experiences a drop-off in protein recovery at higher protein inputs. Magnetic beads may also contaminate samples and instruments, and become costly for larger scale protein preparations. Here, we propose solvent precipitation SP3 (SP4) as an alternative to SP3, omitting magnetic beads and employing brief centrifugation--either with or without low-cost inert glass beads--as the means of aggregated protein capture. SP4 recovered equivalent or greater protein yields for 1-5000 {micro}g preparations and improved reproducibility (median protein R2 0.99 (SP4) vs. 0.97 (SP3)). Deep proteome profiling (n = 9,076) also demonstrated improved recovery by SP4 and a significant enrichment of membrane and low-solubility proteins vs. SP3. The effectiveness of SP4 was verified in three other labs, each confirming equivalent or improved proteome characterisation over SP3. This work suggests that protein precipitation is the primary mechanism of SP3, and reliance on magnetic beads presents protein losses, especially at higher concentrations and amongst hydrophobic proteins. SP4 represents an efficient and effective alternative to SP3, provides the option to omit beads entirely, and offers virtually unlimited scalability of input and volume--all whilst retaining the speed and universality of SP3.

BRIEFSolvent precipitation SP3 (SP4) captures aggregated protein for proteomics sample clean-up by omitting magnetic beads, instead employing brief centrifugation--with or without low-cost inert glass beads. SP4 offers improvements to protein yields, higher reproducibility, and greater recovery of membrane proteins, with verifications from three labs. Protein precipitation appears to be the primary mechanism of SP3, with reliance on magnetic beads presenting protein losses, especially at higher concentrations. SP4 presents an effective alternative to SP3 with improved scalability and equal speed and universality.

GRAPHICAL ABSTRACT

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=135 SRC="FIGDIR/small/461247v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Johnston, H. E.</dc:creator>
<dc:creator>Yadav, K.</dc:creator>
<dc:creator>Kirkpatrick, J. M.</dc:creator>
<dc:creator>Biggs, G. S.</dc:creator>
<dc:creator>Oxley, D.</dc:creator>
<dc:creator>Kramer, H. B.</dc:creator>
<dc:creator>Samant, R. S.</dc:creator>
<dc:date>2021-09-24</dc:date>
<dc:identifier>doi:10.1101/2021.09.24.461247</dc:identifier>
<dc:title><![CDATA[Solvent Precipitation SP3 (SP4) enhances recovery for proteomics sample preparation without magnetic beads]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.03.15.585248v1?rss=1">
<title>
<![CDATA[
Physiologic media renders human iPSC-derived macrophages permissive for M. tuberculosis by rewiring organelle function and metabolism 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.03.15.585248v1?rss=1"
</link>
<description><![CDATA[
In vitro studies are crucial for our understanding of the human macrophage immune functions. However, traditional in vitro culture media poorly reflect the metabolic composition of blood, potentially affecting the outcomes of these studies. Here, we analysed the impact of a physiological medium on human induced pluripotent stem cell (iPSC)-derived macrophages (iPSDM) function. Macrophages cultured in a human plasma-like medium (HPLM) were more permissive to Mycobacterium tuberculosis (Mtb) replication and showed decreased lipid metabolism with increased metabolic polarisation. Functionally, we discovered that HPLM-differentiated macrophages showed different metabolic organelle content and activity. Specifically, HPLM-differentiated macrophages displayed reduced lipid droplet and peroxisome content, increased lysosomal proteolytic activity, and increased mitochondrial activity and dynamics. Inhibiting or inducing lipid droplet formation revealed that lipid droplet content is a key factor influencing macrophage permissiveness to Mtb. These findings underscore the importance of using physiologically relevant media in vitro for accurately studying human macrophage function.
]]></description>
<dc:creator>Bussi, C.</dc:creator>
<dc:creator>Lai, R.</dc:creator>
<dc:creator>Athanasiadi, N.</dc:creator>
<dc:creator>Gutierrez, M. G.</dc:creator>
<dc:date>2024-03-17</dc:date>
<dc:identifier>doi:10.1101/2024.03.15.585248</dc:identifier>
<dc:title><![CDATA[Physiologic media renders human iPSC-derived macrophages permissive for M. tuberculosis by rewiring organelle function and metabolism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.03.15.585102v1?rss=1">
<title>
<![CDATA[
High-resolution genomic ancestry reveals mobility in early medieval Europe 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.03.15.585102v1?rss=1"
</link>
<description><![CDATA[
Ancient DNA has unlocked new genetic histories and shed light on archaeological and historical questions, but many known and unknown historical events have remained below detection thresholds because subtle ancestry changes are challenging to reconstruct. Methods based on sharing of haplotypes1,2 and rare variants3,4 can improve power, but are not explicitly temporal and have not been adopted in unbiased ancestry models. Here, we develop Twigstats, a new approach of time-stratified ancestry analysis that can improve statistical power by an order of magnitude by focusing on coalescences in recent times, while remaining unbiased by population-specific drift. We apply this framework to 1,151 available ancient genomes, focussing on northern and central Europe in the historical period, and show that it allows modelling of individual-level ancestry using preceding genomes and provides previously unavailable resolution to detect broader ancestry transformations. In the first half of the first millennium [~]1-500 CE (Common Era), we observe an expansion of Scandinavian-related ancestry across western, central, and southern Europe. However, in the second half of the millennium [~]500-1000 CE, ancestry patterns suggest the regional disappearance or substantial admixture of these ancestries in multiple regions. Within Scandinavia itself, we document a major ancestry influx by [~]800 CE, when a large proportion of Viking Age individuals carried ancestry from groups related to continental Europe. This primarily affected southern Scandinavia, and was differentially represented in the western and eastern directions of the wider Viking world. We infer detailed ancestry portraits integrated with historical, archaeological, and stable isotope evidence, documenting mobility at an individual level. Overall, our results are consistent with substantial mobility in Europe in the early historical period, and suggest that time-stratified ancestry analysis can provide a new lens for genetic history.
]]></description>
<dc:creator>Speidel, L.</dc:creator>
<dc:creator>Silva, M.</dc:creator>
<dc:creator>Booth, T.</dc:creator>
<dc:creator>Raffield, B.</dc:creator>
<dc:creator>Anastasiadou, K.</dc:creator>
<dc:creator>Barrington, C.</dc:creator>
<dc:creator>Gotherstrom, A.</dc:creator>
<dc:creator>Heather, P.</dc:creator>
<dc:creator>Skoglund, P.</dc:creator>
<dc:date>2024-03-19</dc:date>
<dc:identifier>doi:10.1101/2024.03.15.585102</dc:identifier>
<dc:title><![CDATA[High-resolution genomic ancestry reveals mobility in early medieval Europe]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.03.20.585962v1?rss=1">
<title>
<![CDATA[
mRNA display in cell lysates enables identification of cyclic peptides targeting the BRD3 extraterminal domain 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.03.20.585962v1?rss=1"
</link>
<description><![CDATA[
mRNA display is a powerful technology to screen libraries of >1012 cyclic peptides against a protein target, enabling the rapid discovery of high affinity ligands. These cyclic peptides are particularly well suited to challenging protein targets that have been difficult to drug with small molecules. However, target choice can still be limited as screens are typically performed against purified proteins which often demands the use of isolated domains and precludes the use of aggregation-prone targets. Here, we report a method to perform mRNA display selections in mammalian cell lysates without the need for prior target purification, vastly expanding the potential target scope of mRNA display. We have applied the methodology to identify low to sub-nanomolar peptide binders for two targets; a NanoLuc subunit (LgBiT) and full-length bromodomain-containing protein 3 (BRD3). Our cyclic peptides for BRD3 were found to bind to the extraterminal (ET) domain of BRD3 and the closely related BRD proteins, BRD2 and BRD4. While many chemical probes exist for the bromodomains of BRD proteins, the ET domain is relatively underexplored, making these peptides valuable additions to the BRD toolbox.
]]></description>
<dc:creator>Hurd, C. A.</dc:creator>
<dc:creator>Bush, J.</dc:creator>
<dc:creator>Powell, A. J.</dc:creator>
<dc:creator>Walport, L. J.</dc:creator>
<dc:date>2024-03-20</dc:date>
<dc:identifier>doi:10.1101/2024.03.20.585962</dc:identifier>
<dc:title><![CDATA[mRNA display in cell lysates enables identification of cyclic peptides targeting the BRD3 extraterminal domain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.04.04.588074v1?rss=1">
<title>
<![CDATA[
Caspase cleavage of Influenza A virus M2 disrupts M2-LC3 interaction and regulates virion production 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.04.04.588074v1?rss=1"
</link>
<description><![CDATA[
Influenza A virus (IAV) Matrix 2 (M2) protein is an ion channel, required for efficient viral entry and egress. M2 interacts with the small ubiquitin-like LC3 protein through a cytoplasmic C-terminal LC3 interacting region (LIR). Here, we report that M2 is cleaved by caspases, abolishing the M2-LC3 interaction. A crystal structure of the M2 LIR in complex with LC3 indicates the caspase cleavage tetrapeptide motif (82SAVD85) is an unstructured linear motif that does not overlap with the LIR. Furthermore, an IAV mutant expressing a permanently truncated M2, mimicking caspase cleavage, is impaired in M2 plasma membrane transport and produces a significantly attenuated virus. Our results reveal a dynamic regulation of the M2-LC3 interaction by caspases. This highlights the role of host proteases in regulating IAV exit, relating virion production with host cell state.
]]></description>
<dc:creator>Beale, R.</dc:creator>
<dc:creator>Figueras-Novoa, C.</dc:creator>
<dc:creator>Akutsu, M.</dc:creator>
<dc:creator>Murata, D.</dc:creator>
<dc:creator>Jiang, M.</dc:creator>
<dc:creator>Montaner, B.</dc:creator>
<dc:creator>Dubois, C.</dc:creator>
<dc:creator>Shenoy, A. R.</dc:creator>
<dc:date>2024-04-04</dc:date>
<dc:identifier>doi:10.1101/2024.04.04.588074</dc:identifier>
<dc:title><![CDATA[Caspase cleavage of Influenza A virus M2 disrupts M2-LC3 interaction and regulates virion production]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.03.29.587340v1?rss=1">
<title>
<![CDATA[
HIV-1 Capsid Rapidly Induces Long Lived CPSF6 Puncta in Non-dividing Cells but Similar Puncta Already Exist in Uninfected T-cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.03.29.587340v1?rss=1"
</link>
<description><![CDATA[
The HIV-1 capsid (CA) protein forms the outer shell of the viral core that is released into the cytoplasm upon infection. CA binds various cellular proteins, including CPSF6, that directs HIV-1 integration into speckle associated domains in host chromatin. Upon HIV-1 infection, CPSF6 forms puncta in the nucleus. Here, we characterised these CPSF6 puncta further, in HeLa cells, T-cells and macrophages and confirm that integration and reverse transcription are not required for puncta formation. Indeed, we found that puncta formed very rapidly after infection, correlating with the time that CA entered the nucleus. In aphidicolin-treated HeLa cells and macrophages, puncta were detected for the length of the experiment, suggesting that puncta are only lost upon cell division. CA still co-localised with CPSF6 puncta at the latest time points, considerably after the peak of reverse transcription and integration. Intriguingly, the number of puncta induced in macrophages did not correlate with the MOI, or the total number of nuclear speckles present in each cell suggesting that CA/CPSF6 is only directed to a few nuclear speckles. Furthermore, we found that CPSF6 already co-localised with nuclear speckles in uninfected T-cells suggesting that HIV-1 promotes a natural behaviour of CPSF6.
]]></description>
<dc:creator>Guedan, A.</dc:creator>
<dc:creator>Burley, M.</dc:creator>
<dc:creator>Caroe, E.</dc:creator>
<dc:creator>Bishop, K. N.</dc:creator>
<dc:date>2024-03-29</dc:date>
<dc:identifier>doi:10.1101/2024.03.29.587340</dc:identifier>
<dc:title><![CDATA[HIV-1 Capsid Rapidly Induces Long Lived CPSF6 Puncta in Non-dividing Cells but Similar Puncta Already Exist in Uninfected T-cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.04.01.587366v1?rss=1">
<title>
<![CDATA[
High-throughput screening of human genetic variants by pooled prime editing 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.04.01.587366v1?rss=1"
</link>
<description><![CDATA[
Understanding the effects of rare genetic variants remains challenging, both in coding and non-coding regions. While multiplexed assays of variant effect (MAVEs) have enabled scalable functional assessment of variants, established MAVEs are limited by either exogenous expression of variants or constraints of genome editing. Here, we introduce a pooled prime editing (PE) platform in haploid human cells to scalably assay variants in their endogenous context. We first optimized delivery of variants to HAP1 cells, defining optimal pegRNA designs and establishing a co-selection strategy for improved efficiency. We characterize our platform in the context of negative selection by testing over 7,500 pegRNAs targeting SMARCB1 for editing activity and observing depletion of highly active pegRNAs installing loss-of-function variants. We next assess variants in MLH1 via 6-thioguanine selection, assaying 65.3% of all possible SNVs in a 200-bp region spanning exon 10 and distinguishing LoF variants with high accuracy. Lastly, we assay 362 non-coding MLH1 variants across a 60 kb region in a single experiment, identifying pathogenic variants acting via multiple mechanisms with high specificity. Our analyses detail how filtering for highly active pegRNAs can facilitate both positive and negative selection screens. Accordingly, our platform promises to enable highly scalable functional assessment of human variants.
]]></description>
<dc:creator>Herger, M.</dc:creator>
<dc:creator>Kajba, C. M.</dc:creator>
<dc:creator>Buckley, M.</dc:creator>
<dc:creator>Cunha, A.</dc:creator>
<dc:creator>Strom, M.</dc:creator>
<dc:creator>Findlay, G. M.</dc:creator>
<dc:date>2024-04-01</dc:date>
<dc:identifier>doi:10.1101/2024.04.01.587366</dc:identifier>
<dc:title><![CDATA[High-throughput screening of human genetic variants by pooled prime editing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.04.02.587683v1?rss=1">
<title>
<![CDATA[
HIV-1 Vpr causes separate cell cycle arrests in G2 and M that activate alternative DNA damage pathways 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.04.02.587683v1?rss=1"
</link>
<description><![CDATA[
Vpr is a conserved primate lentiviral accessory protein that induces cell cycle arrest in G2. The precise mechanism of this arrest and its benefit to viral replication is unknown. Here, we show that in addition to G2 arrest, Vpr from HIV-1/SIVcpz and HIV-2 lineages separately induce mitotic arrest through the spindle assembly checkpoint, in contrast to other Vpr proteins that only cause G2 arrest. The G2 arrest was mediated solely by ATR (ataxia telangiectasia and Rad3 related) and this activity caused elevated cellular dNTP levels. The mitotic arrest required ATM (ataxia-telangiectasia mutated) as well as ATR activity and resulted from the formation of HIV-1 Vpr-induced ultra-fine anaphase bridges. Moreover, ectopic expression of the DNA structure-specific endonuclease, MUS81, prevented mitotic but not G2 arrest. Importantly, virion-incorporated Vpr was sufficient to induce cellular changes within 12h post-infection, implying that these events benefit the early stages of HIV infection.

Author SummaryVpr is an accessory protein found in primate lentiviruses. Like other retroviral accessory proteins, it is not absolutely required for viral replication but is thought to overcome a cellular factor that negatively regulates infection. The most well-documented effect of Vpr expression is cell cycle arrest in G2. This has been linked to activation of the DNA damage response (DDR) pathway but there are conflicting reports in the literature as to the mechanism behind this. Here, we show that Vpr from some lentiviruses, in fact, cause two separate cell cycle blocks, in G2 and M, that require different DDR pathways. Other Vpr proteins only cause arrest in G2. Furthermore, we show that degradation of one reported target of Vpr, MUS81, is specifically linked to M but not G2 arrest. This indicates that not all Vpr functions are conserved and helps explain contradictory published results. Additionally, we found that virion-incorporated Vpr protein was able to induce cellular changes, including elevated dNTP levels, within 12 hours of infection suggesting that these events enhance early HIV-1 replication events.
]]></description>
<dc:creator>Hall, R.</dc:creator>
<dc:creator>Ahern, L. M.</dc:creator>
<dc:creator>Yap, M. W.</dc:creator>
<dc:creator>Tsai, M.-H. C.</dc:creator>
<dc:creator>Boucherit, V. C.</dc:creator>
<dc:creator>Takaki, T.</dc:creator>
<dc:creator>Boulton, S. J.</dc:creator>
<dc:creator>Bishop, K. N.</dc:creator>
<dc:date>2024-04-02</dc:date>
<dc:identifier>doi:10.1101/2024.04.02.587683</dc:identifier>
<dc:title><![CDATA[HIV-1 Vpr causes separate cell cycle arrests in G2 and M that activate alternative DNA damage pathways]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.03.26.586830v1?rss=1">
<title>
<![CDATA[
Robust encoding of sub-sniff temporal information in the mouse olfactory bulb 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.03.26.586830v1?rss=1"
</link>
<description><![CDATA[
SummaryThe sensory world is highly dynamic, and the temporal structure of stimuli contains rich information about the environment. Odour plumes are shaped by complex airflow that imprint information about the nature and spatial organisation of the olfactory environment onto their temporal dynamics. Whilst insects and mammals alike can discern high-frequency information, how temporal properties of the olfactory environment are represented in the brain remains largely unknown. Here, we presented temporally rich and systematically varying odour stimuli whilst electrically recording from the output neurons of the mouse olfactory bulb, mitral and tufted cells (MTC). We found that temporal aspects of odour stimuli could readily be read out from MTC responses, with a temporal resolution of up to 20 ms. Remarkably, temporal representation was virtually identical across three different odours. To understand which temporal features are encoded, we developed a single-cell model accurately describing both single-cell and population responses. Temporal receptive fields of MTCs translated between different odours, indicating that MTC tuning to odour quality and dynamics are partially separable. Together, this suggests a stereotypical representation of odour dynamics across projection neurons and can serve as an entry point into dissecting mechanisms underlying how information about the environment is extracted from temporally fluctuating odour plumes.
]]></description>
<dc:creator>Warner, T. P. A.</dc:creator>
<dc:creator>Tootoonian, S.</dc:creator>
<dc:creator>Schaefer, A. T.</dc:creator>
<dc:date>2024-03-29</dc:date>
<dc:identifier>doi:10.1101/2024.03.26.586830</dc:identifier>
<dc:title><![CDATA[Robust encoding of sub-sniff temporal information in the mouse olfactory bulb]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.04.08.588616v1?rss=1">
<title>
<![CDATA[
Plasmodium falciparum protein phosphatase PP7 is required for early ring-stage development 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.04.08.588616v1?rss=1"
</link>
<description><![CDATA[
We previously reported that the Plasmodium falciparum putative serine/threonine protein phosphatase 7 (PP7) is a high confidence substrate of the cAMP-dependent protein kinase (PKA). Here we explore the function of PP7 in asexual P. falciparum blood stage parasites. We show that conditional disruption of PP7 leads to a severe growth arrest. We show that PP7 is a calcium-dependent phosphatase which interacts with calmodulin and calcium-dependent protein kinase 1 (CDPK1), consistent with a role in calcium signalling. Notably, PP7 was found to be dispensable for erythrocyte invasion, but was crucial for ring-stage development, with PP7-null parasites arresting shortly following invasion and showing no transition to ameboid forms. Phosphoproteomic analysis revealed that PP7 may regulate certain PKAc substrates. Its interaction with calmodulin and CDPK1 further emphasise a role in calcium signalling, while its impact on early ring development and PKAc substrate phosphorylation underscores its importance in parasite development.
]]></description>
<dc:creator>Patel, A.</dc:creator>
<dc:creator>Rey, J. A.</dc:creator>
<dc:creator>Flynn, H. R.</dc:creator>
<dc:creator>Kousis, K.</dc:creator>
<dc:creator>Skehel, M. J.</dc:creator>
<dc:creator>Blackman, M.</dc:creator>
<dc:creator>Baker, D. A.</dc:creator>
<dc:date>2024-04-09</dc:date>
<dc:identifier>doi:10.1101/2024.04.08.588616</dc:identifier>
<dc:title><![CDATA[Plasmodium falciparum protein phosphatase PP7 is required for early ring-stage development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.04.10.588783v1?rss=1">
<title>
<![CDATA[
Monolayer force generation and transmission is dictated by focal adhesion distribution 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.04.10.588783v1?rss=1"
</link>
<description><![CDATA[
For tissue development, cells must generate contractile forces which are transmitted to their surrounding matrix or neighbouring cells via adhesion complexes. It is often envisaged that a simple linear counterbalance of cell generated stress with extracellular matrix (ECM) traction forces exists. However, experimental evidence indicates that modulating cell-ECM attachment does not necessarily lead to expected reciprocal changes in intercellular stresses. As ECM composition or mechanical properties are rarely uniform, it is important to understand the complexity of how focal adhesions alter stress transmission and the force-balance of a tissue. To address this, we confined monolayers on adhesive patterns altering focal adhesion distribution. Traction force microscopy and laser ablations of cell-cell junctions were used to examine stresses across epithelial monolayers whilst modulating substrate stiffness. We show that monolayers reach different force-balance states depending on focal adhesion distribution. Using an active matter model and confirmed experimentally, we reveal that a force-balance is generated by non-uniform patterns of cell contractility linked to adhesion patterning. This work highlights the importance of integrating the position of cell-ECM attachments into our vision of the mechanical landscape of living tissues.

TeaserTo infer a tissues force-balance, positional information of focal adhesion distribution needs to be integrated due to the emergence of non-uniform patterns of cell contractility.
]]></description>
<dc:creator>Davis, J. R.</dc:creator>
<dc:creator>Solowiej-Wedderburn, J.</dc:creator>
<dc:creator>Vega, S. L.</dc:creator>
<dc:creator>Burdick, J. A.</dc:creator>
<dc:creator>Dunlop, C.</dc:creator>
<dc:creator>Tapon, N.</dc:creator>
<dc:date>2024-04-10</dc:date>
<dc:identifier>doi:10.1101/2024.04.10.588783</dc:identifier>
<dc:title><![CDATA[Monolayer force generation and transmission is dictated by focal adhesion distribution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.04.10.588848v1?rss=1">
<title>
<![CDATA[
MCM Double Hexamer Loading Visualised with Human Proteins 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.04.10.588848v1?rss=1"
</link>
<description><![CDATA[
Eukaryotic DNA replication begins with the loading of the MCM replicative DNA helicase as a head-to-head double hexamer (DH) at origins of DNA replication1-3. Our current understanding of how DH is assembled by the Origin Recognition Complex (ORC), CDC6 and CDT1 comes mostly from budding yeast. Here we characterise human DH (hDH) loading using biochemical reconstitution and cryo-electron microscopy with purified proteins. We show that hDH engages DNA differently from yeast (yDH), and generates [~]5 base pairs of unwound DNA at the interface between hexamers, as seen in hDH isolated from cells4. We identify several differences from yeast in the order of factor recruitment and dependencies during hDH assembly. Unlike yeast5-8, the ORC6 subunit of ORC is not essential for initial MCM recruitment or hDH loading, but contributes to an alternative hDH assembly pathway requiring an intrinsically disordered region (IDR) in ORC1, which may work through a novel MCM-ORC (hMO*) intermediate. Our work presents a detailed view of how DHs are assembled in an organism utilising sequence-independent replication origins, it provides further evidence for diversity in eukaryotic DH assembly mechanisms9, and it represents the first step toward reconstitution of DNA replication initiation with purified human proteins.
]]></description>
<dc:creator>Weissmann, F.</dc:creator>
<dc:creator>Greiwe, J. F.</dc:creator>
<dc:creator>Pühringer, T.</dc:creator>
<dc:creator>Miller, T. C. R.</dc:creator>
<dc:creator>Diffley, J. F. X.</dc:creator>
<dc:creator>Costa, A.</dc:creator>
<dc:date>2024-04-10</dc:date>
<dc:identifier>doi:10.1101/2024.04.10.588848</dc:identifier>
<dc:title><![CDATA[MCM Double Hexamer Loading Visualised with Human Proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.04.11.588725v1?rss=1">
<title>
<![CDATA[
Spatial proteomic analysis of a lung cancer model reveals regulatory T cells attenuate KRAS-G12C inhibitor-induced immune responses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.04.11.588725v1?rss=1"
</link>
<description><![CDATA[
We recently showed that lung tumor specific KRAS-G12C inhibition causes remodelling of the tumor immune microenvironment from cold to hot. As a result, KRAS-G12C inhibition is able to synergise with anti-PD-1 treatment, but only in tumor models that were already moderately responsive to immune checkpoint blockade at baseline. To investigate mechanisms that restrain immunotherapy sensitivity in non-responsive tumors, we used multiplex imaging mass cytometry to explore spatial patterns in the tumor microenvironment of the highly immune evasive KRAS mutant murine Lewis Lung Cancer model. Clustering of close neighbour information per cell allowed characterisation of spatial patterns or  communities in the tissue. We identified a community harbouring features of localised T-cell activation, where CD4+ and CD8+ T cells and dendritic cells were gathered together. KRAS-G12C inhibition led to increased expression of PD-1 on T cells, CXCL9 expression by dendritic cells, together with increased proliferation and potential cytotoxicity of CD8+ T cells, indicating an effector response. However, we also observed a high incidence of regulatory T cells (Tregs) within this community, which had frequent contact with effector T cells, suggesting that Tregs may be able to dampen anti-tumoral immune responses following KRAS-G12C inhibition. Similar communities were detected in human lung adenocarcinoma clinical samples. Depleting Tregs in vivo with anti-CTLA-4 antibody rescued the anti-tumor immune response and led to enhanced tumor control in combination with anti-PD-1 and KRAS-G12C inhibitor. We therefore propose use of KRAS-G12C inhibitor in combination with Treg depletion as a therapeutic opportunity that increases anti-tumoral immune responses and initiates tumor regression.

One sentence summarySpatial analysis identified regulatory T cells as potential source of local T cell repression, mediating resistance to KRAS-G12Ci and anti-PD1 therapy.
]]></description>
<dc:creator>Cole, M.</dc:creator>
<dc:creator>Anastasiou, P.</dc:creator>
<dc:creator>Lee, C.</dc:creator>
<dc:creator>Moore, C.</dc:creator>
<dc:creator>Mugarza, E.</dc:creator>
<dc:creator>Jones, M.</dc:creator>
<dc:creator>Valand, K.</dc:creator>
<dc:creator>Rana, S.</dc:creator>
<dc:creator>Colliver, E.</dc:creator>
<dc:creator>Angelova, M.</dc:creator>
<dc:creator>Enfield, K.</dc:creator>
<dc:creator>Magness, A.</dc:creator>
<dc:creator>Mullokandov, A.</dc:creator>
<dc:creator>Kelly, G.</dc:creator>
<dc:creator>de Gruijl, T.</dc:creator>
<dc:creator>Molina-Arcas, M.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Downward, J.</dc:creator>
<dc:creator>van Maldegem, F.</dc:creator>
<dc:date>2024-04-13</dc:date>
<dc:identifier>doi:10.1101/2024.04.11.588725</dc:identifier>
<dc:title><![CDATA[Spatial proteomic analysis of a lung cancer model reveals regulatory T cells attenuate KRAS-G12C inhibitor-induced immune responses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.04.12.585865v1?rss=1">
<title>
<![CDATA[
Mutations in the IgG B cell receptor associated with class-switched B cell lymphomas 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.04.12.585865v1?rss=1"
</link>
<description><![CDATA[
Immunoglobulin class-switching from IgM to IgG enhances B cell receptor (BCR) signalling1,2 and promotes germinal centre (GC) B cell responses to antigens3,4. In contrast, non-Hodgkin lymphomas derived from GC B cells typically avoid IgG BCR expression and retain the unswitched IgM BCR, suggesting that the IgG BCR may protect B cells from malignant transformation5,6. However, the mechanism of this phenomenon and its significance for the pathogenicity of IgG-expressing lymphomas remains unclear. Here, we report that IgG-positive follicular lymphoma (FL) and the related EZB subset of diffuse large B cell lymphoma (DLBCL) acquire mutations in the IgG heavy chain, disrupting its unique intracellular tail. Enforced class switching of IgM-expressing EZB DLBCL cell lines to IgG reduces BCR surface levels, signalling via phosphoinositide-3 kinase (PI3K), levels of MYC, cell proliferation and in vivo growth. Inhibiting GSK3, a target of BCR-PI3K signalling, or stimulating the BCR rescues IgG+ cell proliferation. In contrast, IgG tail-truncating mutations enhance BCR surface expression, intracellular signalling and competitive growth. These findings suggest that the expansion of IgG-switched GC-like B lymphoma cells is limited by low tonic PI3K activity of the wild-type IgG BCR, but a subset of these cancers acquires mutations of the IgG intracellular tail that reverse this effect, promoting the oncogenicity of their BCRs. The presence of IgG tail mutations underscores the importance of isotype-specific BCR signalling in the pathogenesis of FL and EZB DLBCL and can potentially inform therapeutic targeting with BCR signalling inhibitors or antibody-drug conjugates.
]]></description>
<dc:creator>Wasim, L.</dc:creator>
<dc:creator>Chu, S. W. T.</dc:creator>
<dc:creator>Sale, B.</dc:creator>
<dc:creator>Pickard, L.</dc:creator>
<dc:creator>Leonard, S.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Tolarova, H.</dc:creator>
<dc:creator>Tean, Z. S.</dc:creator>
<dc:creator>Engels, N.</dc:creator>
<dc:creator>Calado, D. P.</dc:creator>
<dc:creator>Tarte, K.</dc:creator>
<dc:creator>Okosun, J.</dc:creator>
<dc:creator>Forconi, F.</dc:creator>
<dc:creator>Tolar, P.</dc:creator>
<dc:date>2024-04-13</dc:date>
<dc:identifier>doi:10.1101/2024.04.12.585865</dc:identifier>
<dc:title><![CDATA[Mutations in the IgG B cell receptor associated with class-switched B cell lymphomas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.11.23.568045v1?rss=1">
<title>
<![CDATA[
Detection and spread of high pathogenicity avian influenza virus H5N1 in the Antarctic Region 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.11.23.568045v1?rss=1"
</link>
<description><![CDATA[
Until recent events, the Antarctic was the only major geographical region in which high pathogenicity avian influenza virus (HPAIV) had never previously been detected. The current panzootic of H5N1 HPAIV has decimated wild bird populations across Europe, North America, and South America. Here we report on the detection of clade 2.3.4.4b H5N1 HPAIV in the Antarctic and sub-Antarctic regions of South Georgia and the Falkland Islands, respectively. We initially detected H5N1 HPAIV in samples collected from brown skuas at Bird Island, South Georgia on 8th October 2023. Since this detection, mortalities were observed in brown skuas, kelp gulls, South Georgia shag, Antarctic tern, elephant seals and fur seals at multiple sites across South Georgia. Subsequent testing confirmed H5N1 HPAIV across several sampling locations in multiple avian species and two seal species. Simultaneously, we also confirmed H5N1 HPAIV in southern fulmar and black-browed albatross in the Falkland Islands. Genetic assessment of the virus indicates spread from South America, likely through movement of migratory birds. Critically, genetic assessment of sequences from mammalian species demonstrates no increased risk to human populations. Here we describe the detection, species impact and genetic composition of the virus and propose both introductory routes and potential long-term impact on avian and mammalian species across the Antarctic region. We also speculate on the threat to specific populations following recent reports in the area.
]]></description>
<dc:creator>Bennison, A.</dc:creator>
<dc:creator>Byrne, A. M.</dc:creator>
<dc:creator>Reid, S. M.</dc:creator>
<dc:creator>Lynton-Jenkins, J. G.</dc:creator>
<dc:creator>Mollett, B.</dc:creator>
<dc:creator>De Sliva, D.</dc:creator>
<dc:creator>Peers-Dent, J.</dc:creator>
<dc:creator>Finlayson, K. E.</dc:creator>
<dc:creator>Hall, R.</dc:creator>
<dc:creator>Blockley, F.</dc:creator>
<dc:creator>Blyth, M.</dc:creator>
<dc:creator>Falchieri, M.</dc:creator>
<dc:creator>Fowler, Z.</dc:creator>
<dc:creator>Fitzcharles, E.</dc:creator>
<dc:creator>Brown, I. H.</dc:creator>
<dc:creator>James, J.</dc:creator>
<dc:creator>Banyard, A. C.</dc:creator>
<dc:date>2023-11-24</dc:date>
<dc:identifier>doi:10.1101/2023.11.23.568045</dc:identifier>
<dc:title><![CDATA[Detection and spread of high pathogenicity avian influenza virus H5N1 in the Antarctic Region]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.04.17.589864v1?rss=1">
<title>
<![CDATA[
The cis-regulatory logic integrating spatial and temporal patterning in the vertebrate neural tube 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.04.17.589864v1?rss=1"
</link>
<description><![CDATA[
The vertebrate neural tube generates a large diversity of molecularly and functionally distinct neurons and glia from a small progenitor pool. While the role of spatial patterning in organising cell fate specification has been extensively studied, temporal patterning, which controls the timing of cell type generation, is equally important. Here we define a global temporal programme operating in progenitors throughout the vertebrate nervous systems that governs cell fate choices by regulating chromatin accessibility. Perturbation of this cis-regulatory programme affects sequential cell fate transitions in neural progenitors and the identity of their progeny. The temporal programme operates in parallel to spatial patterning, ensuring the timely availability of regulatory elements for spatial determinants to direct cell-type specific gene expression. These findings identify a chronotopic spatiotemporal integration strategy in which a global temporal chromatin programme determines the output of a spatial gene regulatory network resulting in the temporally and spatially ordered allocation of cell type identity.
]]></description>
<dc:creator>Zhang, I.</dc:creator>
<dc:creator>Boezio, G. L.</dc:creator>
<dc:creator>Cornwall-Scoones, J.</dc:creator>
<dc:creator>Frith, T.</dc:creator>
<dc:creator>Jiang, M.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>Lovell-Badge, R.</dc:creator>
<dc:creator>Sagner, A.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Delas, M. J.</dc:creator>
<dc:date>2024-04-17</dc:date>
<dc:identifier>doi:10.1101/2024.04.17.589864</dc:identifier>
<dc:title><![CDATA[The cis-regulatory logic integrating spatial and temporal patterning in the vertebrate neural tube]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.04.17.589886v1?rss=1">
<title>
<![CDATA[
Accurate evaluation of live-virus microneutralisation for SARS-CoV-2 variant JN.1 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.04.17.589886v1?rss=1"
</link>
<description><![CDATA[
Emerging SARS-CoV-2 variants require rapid assessments of pathogenicity and evasion of existing immunity to inform policy. A crucial component of these assessments is accurate estimation of serum neutralising antibody titres using cultured live virus isolates. Here, we report our updated culture methods for Omicron sub-variant JN.1 using Caco-2 cells and the subsequent evaluation of neutralising antibody titres (nAbTs) in recipients of BNT162b2-XBB.1.5 monovalent and the Ancestral/BA.5 containing bivalent vaccines. We compared culture of JN.1 in either Vero V1 cells or Caco-2 cells, finding culture in Vero V1 either resulted in low-titre stocks or induced crucial mutations at the Spike furin cleavage site. Using the sequence-clean culture stocks generated in Caco-2 cells, we assessed serum samples from 71 healthy adults eligible for a COVID-19 vaccination given as a 5th dose booster: all participants had detectable nAbs against JN.1 prior to vaccination, with baseline/pre-existing nAbTs between both vaccine groups comparable (p = 0.240). However, nAbTs against JN.1 post-vaccination were 2.6-fold higher for recipients of the monovalent XBB1.5 vaccine than the BA.4/5 bivalent vaccine (p<0.001). Regular re-appraisal of methods involved in the evaluation of new variants is required to ensure robust data are used to underpin crucial severity assessments as variants arise and vaccine strain selection decisions.
]]></description>
<dc:creator>Dowgier, G.</dc:creator>
<dc:creator>Hobbs, A.</dc:creator>
<dc:creator>Greenwood, D.</dc:creator>
<dc:creator>Shawe-Taylor, M.</dc:creator>
<dc:creator>Stevenson-Leggett, P.</dc:creator>
<dc:creator>Bazire, J.</dc:creator>
<dc:creator>Penn, R.</dc:creator>
<dc:creator>Harvey, R.</dc:creator>
<dc:creator>Libri, V.</dc:creator>
<dc:creator>Kassiotis, G.</dc:creator>
<dc:creator>Gamblin, S.</dc:creator>
<dc:creator>Lewis, N. S.</dc:creator>
<dc:creator>Williams, B.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Bauer, D. L.</dc:creator>
<dc:creator>Carr, E. J.</dc:creator>
<dc:creator>Wall, E. C.</dc:creator>
<dc:creator>Wu, M. Y.</dc:creator>
<dc:date>2024-04-18</dc:date>
<dc:identifier>doi:10.1101/2024.04.17.589886</dc:identifier>
<dc:title><![CDATA[Accurate evaluation of live-virus microneutralisation for SARS-CoV-2 variant JN.1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.04.18.590032v1?rss=1">
<title>
<![CDATA[
Rapid mortality in captive bush dogs (Speothos venaticus) caused by influenza A of avian origin (H5N1) at a wildlife collection in the United Kingdom 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.04.18.590032v1?rss=1"
</link>
<description><![CDATA[
Europe has suffered unprecedented epizootics of high pathogenicity avian influenza (HPAI) clade 2.3.4.4b H5N1 since Autumn 2021. As well as impacting upon commercial and wild avian species, the virus has also infected mammalian species more than ever observed previously. Mammalian species involved in spill over events have primarily been scavenging terrestrial carnivores and farmed mammalian species although marine mammals have also been affected. Alongside reports of detections in mammalian species found dead through different surveillance schemes, several mass mortality events have been reported in farmed and wild animals. During November 2022, an unusual mortality event was reported in captive bush dogs (Speothos venaticus) with clade 2.3.4.4b H5N1 HPAIV of avian origin being the causative agent. The event involved an enclosure of fifteen bush dogs, ten of which succumbed during a nine-day period with some dogs exhibiting neurological disease. Ingestion of infected meat is proposed as the most likely infection route.
]]></description>
<dc:creator>Falchieri, M.</dc:creator>
<dc:creator>Reid, S.</dc:creator>
<dc:creator>Dastderji, A.</dc:creator>
<dc:creator>Cracknell, J.</dc:creator>
<dc:creator>Warren, C. J.</dc:creator>
<dc:creator>Mollett, B.</dc:creator>
<dc:creator>Peers-Dent, J.</dc:creator>
<dc:creator>Schlachter, A.-L.</dc:creator>
<dc:creator>Mcginn, N.</dc:creator>
<dc:creator>Hepple, R.</dc:creator>
<dc:creator>Thomas, S.</dc:creator>
<dc:creator>Ridout, S.</dc:creator>
<dc:creator>Quayle, J.</dc:creator>
<dc:creator>Pizzi, R.</dc:creator>
<dc:creator>Nunez, A.</dc:creator>
<dc:creator>Byrne, A. M. P.</dc:creator>
<dc:creator>James, J.</dc:creator>
<dc:creator>Banyard, A. C.</dc:creator>
<dc:date>2024-04-18</dc:date>
<dc:identifier>doi:10.1101/2024.04.18.590032</dc:identifier>
<dc:title><![CDATA[Rapid mortality in captive bush dogs (Speothos venaticus) caused by influenza A of avian origin (H5N1) at a wildlife collection in the United Kingdom]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.04.19.590313v1?rss=1">
<title>
<![CDATA[
Spherical Phenotype Clustering 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.04.19.590313v1?rss=1"
</link>
<description><![CDATA[
Phenotypic screening experiments produce many microscope images of cells under diverse perturbations, with biologically significant responses often subtle or difficult to identify visually. A central challenge is to extract image representations that distinguish activity from controls and group phenotypically similar perturbations. In this work we propose new adaptations of contrastive loss functions that incorporate experimental metadata as learned class vectors, and a geometrically inspired variant, called SPC, where class vectors are confined to the unit sphere and updated only by attractive terms (allowing more overlap of phenotypically similar classes). The approach is tested on two popular benchmarking datasets, BBBC021 and RxRx3-core; and we also evaluate performance on uncurated screens of HaCaT cells to gauge effectiveness in a realistic use-case scenario. We find we outperform prior methods across the three datasets and on a wide array of metrics measuring phenotype grouping, biological recall, drug-target interaction and mechanism-of-action inference. We also show we maintain this improved performance compared to models over 10x larger in parameter count, and that SPC can be used as an effective fine-tuning technique. The method is easy to implement and is well suited to settings with limited data or compute resources.
]]></description>
<dc:creator>Nightingale, L.</dc:creator>
<dc:creator>Tuersley, J.</dc:creator>
<dc:creator>Cairoli, A.</dc:creator>
<dc:creator>Howes, J.</dc:creator>
<dc:creator>Powell, A.</dc:creator>
<dc:creator>Green, D.</dc:creator>
<dc:creator>Strange, A.</dc:creator>
<dc:creator>Warchal, S.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:date>2024-04-25</dc:date>
<dc:identifier>doi:10.1101/2024.04.19.590313</dc:identifier>
<dc:title><![CDATA[Spherical Phenotype Clustering]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.04.29.591629v1?rss=1">
<title>
<![CDATA[
Spatiotemporal Orchestration of Mitosis by Cyclin-Dependent Kinase 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.04.29.591629v1?rss=1"
</link>
<description><![CDATA[
Mitotic onset is a critical transition for eukaryotic cell proliferation. The prevailing view for its control is that cyclin-dependent kinase (CDK) is activated first in the cytoplasm, at the centrosome, which then initiates mitosis1-3. Bistability in CDK activation ensures the transition is irreversible but how this unfolds in a spatially compartmentalized cell is unknown4-8. Here using fission yeast, we show that CDK is actually activated in the nucleus first, not the cytoplasm, and that the bistable responses dramatically differ within the nucleus and cytoplasm. There is a stronger response in the nucleus permitting mitotic signal propagation from there to the cytoplasm. Abolishing cyclin-CDK localization to the yeast centrosome led to activation occurring only in the nucleus, spatially uncoupling the nucleus and cytoplasm mitotically, suggesting centrosomal cyclin-CDK acts as a "signal relayer". We propose that the key mitotic regulatory system operates in the nucleus in proximity to DNA, allowing incomplete DNA replication and DNA damage to be effectively monitored to preserve genome integrity and for integration of ploidy within the CDK control network. This spatiotemporal regulatory framework establishes core principles for control of the onset of mitosis in vivo, which will help inform how CDK controls mitotic onset in other eukaryotes.
]]></description>
<dc:creator>Kapadia, N.</dc:creator>
<dc:creator>Nurse, P.</dc:creator>
<dc:date>2024-04-30</dc:date>
<dc:identifier>doi:10.1101/2024.04.29.591629</dc:identifier>
<dc:title><![CDATA[Spatiotemporal Orchestration of Mitosis by Cyclin-Dependent Kinase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.04.30.591261v1?rss=1">
<title>
<![CDATA[
The Streptococcus pneumoniae transcriptome in patient cerebrospinal fluid identifies novel virulence factors required for meningitis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.04.30.591261v1?rss=1"
</link>
<description><![CDATA[
To better understand Streptococcus pneumoniae pathogenesis we performed RNA sequencing on cerebrospinal fluid (CSF) from meningitis patients to identify bacterial genes expressed during invasion of the central nervous system. Comparison to transcriptome data for serotype 1 S. pneumoniae cultured in ex vivo human CSF defined a subset of 57 genes with high expression during human meningitis. Deletion of two of the most highly expressed genetic loci, bgaA (encodes for a {beta}-galactosidase) or the SP_1801-5 putative stress response operon, resulted in S. pneumoniae strains still able to transmigrate the blood brain barrier but which were more susceptible to complement opsonisation and unable to maintain brain infection in a murine meningitis model. In 1144 meningitis patients, infection with bgaA containing S. pneumoniae strains was associated with a higher mortality (22% versus 14% p=0.02). These data demonstrate that direct bacterial RNAseq from CSF can identify previously undescribed S. pneumoniae virulence factors required for meningitis pathogenesis.
]]></description>
<dc:creator>Wall, E. C.</dc:creator>
<dc:creator>Guerra-Assuncao, J.-A.</dc:creator>
<dc:creator>Yang, M.</dc:creator>
<dc:creator>Konig, R.</dc:creator>
<dc:creator>Audshasai, T.</dc:creator>
<dc:creator>Proust, A.</dc:creator>
<dc:creator>Aprianto, R.</dc:creator>
<dc:creator>Ramos-Sevillano, E.</dc:creator>
<dc:creator>Ercoli, G.</dc:creator>
<dc:creator>Bordin, N.</dc:creator>
<dc:creator>Agostinho-Terra, V. S.</dc:creator>
<dc:creator>Veening, J.-W.</dc:creator>
<dc:creator>Lalloo, D. G.</dc:creator>
<dc:creator>Wren, B. W.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Brouwer, M. C.</dc:creator>
<dc:creator>van de Beek, D.</dc:creator>
<dc:creator>Kadiloglu, A.</dc:creator>
<dc:creator>Heyderman, R. S.</dc:creator>
<dc:creator>Brown, J. S.</dc:creator>
<dc:date>2024-04-30</dc:date>
<dc:identifier>doi:10.1101/2024.04.30.591261</dc:identifier>
<dc:title><![CDATA[The Streptococcus pneumoniae transcriptome in patient cerebrospinal fluid identifies novel virulence factors required for meningitis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.04.29.591666v1?rss=1">
<title>
<![CDATA[
Addressing pandemic-wide systematic errors in the SARS-CoV-2 phylogeny 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.04.29.591666v1?rss=1"
</link>
<description><![CDATA[
The SARS-CoV-2 genome occupies a unique place in infection biology - it is the most highly sequenced genome on earth (making up over 20% of public sequencing datasets) with fine scale information on sampling date and geography, and has been subject to unprecedented intense analysis. As a result, these phylogenetic data are an incredibly valuable resource for science and public health. However, the vast majority of the data was sequenced by tiling amplicons across the full genome, with amplicon schemes that changed over the pandemic as mutations in the viral genome interacted with primer binding sites. In combination with the disparate set of genome assembly workflows and lack of consistent quality control (QC) processes, the current genomes have many systematic errors that have evolved with the virus and amplicon schemes. These errors have significant impacts on the phylogeny, and therefore over the last few years, many thousands of hours of researchers time has been spent in "eyeballing" trees, looking for artefacts, and then patching the tree.

Given the huge value of this dataset, we therefore set out to reprocess the complete set of public raw sequence data in a rigorous amplicon-aware manner, and build a cleaner phylogeny. Here we provide a global tree of 4,471,579 samples, built from a consistently assembled set of high quality consensus sequences from all available public data as of June 2024, viewable at https://viridian.taxonium.org. Each genome was constructed using a novel assembly tool called Viridian (https://github.com/iqbal-lab-org/viridian), developed specifically to process amplicon sequence data, eliminating artefactual errors and mask the genome at low quality positions. We provide simulation and empirical validation of the methodology, and quantify the improvement in the phylogeny. We hope the tree, consensus sequences and Viridian will be a valuable resource for researchers.
]]></description>
<dc:creator>Hunt, M.</dc:creator>
<dc:creator>Hinrichs, A. S.</dc:creator>
<dc:creator>Anderson, D.</dc:creator>
<dc:creator>Karim, L.</dc:creator>
<dc:creator>Dearlove, B. L.</dc:creator>
<dc:creator>Knaggs, J.</dc:creator>
<dc:creator>Constantinides, B.</dc:creator>
<dc:creator>Fowler, P. W.</dc:creator>
<dc:creator>Rodger, G.</dc:creator>
<dc:creator>Street, T. L.</dc:creator>
<dc:creator>Lumley, S. F.</dc:creator>
<dc:creator>Webster, H.</dc:creator>
<dc:creator>Sanderson, T.</dc:creator>
<dc:creator>Ruis, C.</dc:creator>
<dc:creator>De Maio, N.</dc:creator>
<dc:creator>Amenga-Etego, L. N.</dc:creator>
<dc:creator>Amuzu, D. S.</dc:creator>
<dc:creator>Avaro, M.</dc:creator>
<dc:creator>Awandare, G. A.</dc:creator>
<dc:creator>Ayivor-Djanie, R.</dc:creator>
<dc:creator>Bashton, M.</dc:creator>
<dc:creator>Batty, E. M.</dc:creator>
<dc:creator>Bediako, Y.</dc:creator>
<dc:creator>De Belder, D.</dc:creator>
<dc:creator>Benedetti, E.</dc:creator>
<dc:creator>Bergthaler, A.</dc:creator>
<dc:creator>Boers, S. A.</dc:creator>
<dc:creator>Campos, J.</dc:creator>
<dc:creator>Carr, R. A. A.</dc:creator>
<dc:creator>Cuba, F.</dc:creator>
<dc:creator>Dattero, M. E.</dc:creator>
<dc:creator>Dejnirattisai, W.</dc:creator>
<dc:creator>Dilthey, A. T.</dc:creator>
<dc:creator>Duedu, K. O.</dc:creator>
<dc:creator>Endler, L.</dc:creator>
<dc:creator>Engelmann, I.</dc:creator>
<dc:creator>Francisco, N. M.</dc:creator>
<dc:creator>Fuchs, J.</dc:creator>
<dc:creator>Gnimpieba Z., E.</dc:creator>
<dc:creator>Groc, S.</dc:creator>
<dc:creator>Gyamfi, J.</dc:creator>
<dc:creator>Heemskerk,</dc:creator>
<dc:date>2024-04-30</dc:date>
<dc:identifier>doi:10.1101/2024.04.29.591666</dc:identifier>
<dc:title><![CDATA[Addressing pandemic-wide systematic errors in the SARS-CoV-2 phylogeny]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.16.594506v1?rss=1">
<title>
<![CDATA[
An in vivo CRISPR screen in chick embryos reveals a role for MLLT3 in specification of neural cells from the caudal epiblast 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.16.594506v1?rss=1"
</link>
<description><![CDATA[
Tissue development relies on the coordinated differentiation of stem cells in dynamically changing environments. The formation of the vertebrate neural tube from stem cells in the caudal lateral epiblast (CLE) is a well characterized example. Despite an understanding of the signalling pathways involved, the gene regulatory mechanisms remain poorly defined. To address this, we developed a multiplexed in vivo CRISPR screening approach in chick embryos targeting genes expressed in the caudal epiblast and neural tube. This revealed a role for MLLT3, a component of the super elongation complex, in the specification of neural fate. Perturbation of MLLT3 disrupted neural tube morphology and reduced neural fate acquisition. Mutant forms of Retinoic Acid Receptor A lacking the MLLT3 binding domain similarly reduced neural fate acquisition. Together, these findings validate an in vivo CRISPR screen strategy in chick embryos and identify a previously unreported role for MLLT3 in caudal neural tissue specification.
]]></description>
<dc:creator>Libby, A. R.</dc:creator>
<dc:creator>Rito, T.</dc:creator>
<dc:creator>Radley, A.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:date>2024-05-16</dc:date>
<dc:identifier>doi:10.1101/2024.05.16.594506</dc:identifier>
<dc:title><![CDATA[An in vivo CRISPR screen in chick embryos reveals a role for MLLT3 in specification of neural cells from the caudal epiblast]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.12.593776v1?rss=1">
<title>
<![CDATA[
Discovery and Characterization of a Chemical Probe for Cyclin-Dependent Kinase-Like 2 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.12.593776v1?rss=1"
</link>
<description><![CDATA[
Acylaminoindazole-based inhibitors of CDKL2 were identified via analyses of cell-free binding and selectivity data. Compound 9 was selected as a CDKL2 chemical probe based on its potent inhibition of CDKL2 enzymatic activity, engagement of CDKL2 in cells, and excellent kinome-wide selectivity, especially when used in cells. Compound 16 was designed as a negative control to be used alongside compound 9 in experiments to interrogate CDKL2-mediated biology. A solved co-crystal structure of compound 9 bound to CDKL2 highlighted key interactions it makes within its ATP-binding site. Inhibition of downstream phosphorylation of EB2, a CDKL2 substrate, in rat primary neurons provided evidence that engagement of CDKL2 by compound 9 in cells resulted in inhibition of its activity. When used at relevant concentrations, compound 9 does not impact the viability of rat primary neurons or certain breast cancer cells nor elicit consistent changes in the expression of proteins involved in epithelial-mesenchymal transition.
]]></description>
<dc:creator>Bashore, F. M.</dc:creator>
<dc:creator>Min, S. M.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Howell, S.</dc:creator>
<dc:creator>Rinderle, C. H.</dc:creator>
<dc:creator>Morel, G.</dc:creator>
<dc:creator>Silvaroli, J. A.</dc:creator>
<dc:creator>Wells, C. I.</dc:creator>
<dc:creator>Bunnell, B. A.</dc:creator>
<dc:creator>Drewry, D.</dc:creator>
<dc:creator>Pabla, N. S.</dc:creator>
<dc:creator>Ultanir, S. K.</dc:creator>
<dc:creator>Bullock, A. N.</dc:creator>
<dc:creator>Axtman, A. D.</dc:creator>
<dc:date>2024-05-14</dc:date>
<dc:identifier>doi:10.1101/2024.05.12.593776</dc:identifier>
<dc:title><![CDATA[Discovery and Characterization of a Chemical Probe for Cyclin-Dependent Kinase-Like 2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.14.594112v1?rss=1">
<title>
<![CDATA[
Imaging of MAP kinase dynamics reveals endocytic regulation of pulsatile signalling and network re-wiring in response to targeted therapy in EGFR-mutant non-small cell lung cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.14.594112v1?rss=1"
</link>
<description><![CDATA[
A better understanding of the signalling mechanisms underlying transitions from drug-sensitive to drug-tolerant states is required to overcome therapy failure. We combined single-cell biosensor imaging with functional perturbations to investigate the regulation of oncogenic signalling in EGFR-mutant lung adenocarcinoma. We find that despite the constant presence of the mutant oncogene, ERK signalling exhibits pulsatile dynamics, with pulse characteristics determined by the endocytic machinery. Analysis of drug-tolerant persisters (DTPs) revealed that, after an initial phase of complete pathway shut-down, signalling was rewired leading to renewed ERK pulses that drive cell cycle progression. FAK- and SRC-regulated adhesion complexes replace mutant EGFR as the driver of reactivated ERK pulses in DTPs, yet they remain controlled by the membrane trafficking machinery. We show that DTPs rely on additional survival pathways including YAP signalling, and that the phosphatase PTPRS represents a key node in therapy resistant cells, coordinating regulation of ERK, the cytoskeleton, and YAP.
]]></description>
<dc:creator>Le Marois, A. M. I.</dc:creator>
<dc:creator>Bailey, S.</dc:creator>
<dc:creator>Hooper, S.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Sparks, H.</dc:creator>
<dc:creator>Alexandrov, Y.</dc:creator>
<dc:creator>Caswell, D. R.</dc:creator>
<dc:creator>Fröhlich, F.</dc:creator>
<dc:creator>Schlegelmilch, K.</dc:creator>
<dc:creator>Valand, K.</dc:creator>
<dc:creator>Martin, M. J.</dc:creator>
<dc:creator>Narvaez, A. J.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Downward, J.</dc:creator>
<dc:creator>Dunsby, C.</dc:creator>
<dc:creator>French, P. M.</dc:creator>
<dc:creator>Sahai, E.</dc:creator>
<dc:date>2024-05-17</dc:date>
<dc:identifier>doi:10.1101/2024.05.14.594112</dc:identifier>
<dc:title><![CDATA[Imaging of MAP kinase dynamics reveals endocytic regulation of pulsatile signalling and network re-wiring in response to targeted therapy in EGFR-mutant non-small cell lung cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.20.594983v1?rss=1">
<title>
<![CDATA[
p53 and TIGAR promote redox control to protect against metabolic dysfunction-associated steatohepatitis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.20.594983v1?rss=1"
</link>
<description><![CDATA[
TP53 is a potent tumour suppressor that coordinates diverse stress response programmes within the cell. The activity of p53 is frequently context and cell type-dependent, and ranges from pro-survival activities, including the implementation of transient cell cycle arrest and metabolic rewiring, through to cell death. In addition to tumour suppressor functions, p53 also has established roles in the pathological response to stress that occurs during tissue damage and repair, including within the liver. Metabolic dysfunction-associated steatohepatitis (MASH) is a major driver of hepatocellular carcinoma (HCC), but our understanding of the molecular determinants of MASH development remains incomplete.

Here, using a p53 reporter mouse, we report early and sustained activation of hepatic p53 in response to an obesogenic high fat and high sugar diet. In this context, liver-specific loss of p53 accelerates the progression of benign fatty liver disease to MASH that is characterised by high levels of reactive oxygen species (ROS), extensive fibrosis, and chronic inflammation. Using an in vitro culture system, we show that p53 functions to control ROS and protect against the development of MASH, in part through induction of the antioxidant gene TP53-induced glycolysis and apoptosis regulator (TIGAR). Our work demonstrates an important role for the p53-TIGAR axis in protecting against MASH, and identifies redox control as an essential barrier against liver disease progression.
]]></description>
<dc:creator>Wittke, C. I.</dc:creator>
<dc:creator>Cheung, E. C.</dc:creator>
<dc:creator>Athineos, D.</dc:creator>
<dc:creator>Clements, N.</dc:creator>
<dc:creator>Butler, L.</dc:creator>
<dc:creator>Hughes, M.</dc:creator>
<dc:creator>Morrison, V.</dc:creator>
<dc:creator>Watt, D.</dc:creator>
<dc:creator>Blyth, K.</dc:creator>
<dc:creator>Vousden, K.</dc:creator>
<dc:creator>Humpton, T. J.</dc:creator>
<dc:date>2024-05-21</dc:date>
<dc:identifier>doi:10.1101/2024.05.20.594983</dc:identifier>
<dc:title><![CDATA[p53 and TIGAR promote redox control to protect against metabolic dysfunction-associated steatohepatitis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.18.594804v1?rss=1">
<title>
<![CDATA[
Sympathetic neuron-derived NPY protects from obesity by sustaining the mural progenitors of thermogenic adipocytes. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.18.594804v1?rss=1"
</link>
<description><![CDATA[
Neuropeptide Y (NPY) is secreted by sympathetic nerves1,2, but its direct impact on thermogenic adipocytes is unknown. Here we uncover the mechanism by which peripheral NPY protects from obesity. Our imaging of cleared murine brown and white adipose tissue (BAT and WAT) established that NPY+ sympathetic axons are only a minority that mostly maps to the peri-vasculature; our analysis of single-cell RNA-sequencing datasets identifies mural cells as the main NPY-responsive cells in adipose tissues. We show that NPY sustains mural cells, which are known to be a source of beige cells in both BAT and WAT3-5 and that NPY facilitates the differentiation to thermogenic adipocytes. We found that diet-induced-obesity leads to neuropathy of NPY+ axons and concomitant depletion of the mural cell pool of beige fat progenitors. This defect is replicated in conditional knockout (cKO) mice with NPY specifically abrogated from sympathetic neurons. These cKO mice have whitened BAT with reduced thermogenic ability and lower energy expenditure even before the onset of obesity; they develop adult-onset obesity on a regular chow diet and are more susceptible to diet induced obesity without increasing food consumption. Our results indicate that, relative to central NPY, peripheral NPY produced by the sympathetic nerves has the opposite effect on body weight homeostasis by sustaining the proliferation of the mural cell progenitors of thermogenic adipocytes.
]]></description>
<dc:creator>Zhu, Y.</dc:creator>
<dc:creator>Yao, L.</dc:creator>
<dc:creator>Gallo-Ferraz, A. L.</dc:creator>
<dc:creator>Bombassaro, B.</dc:creator>
<dc:creator>Simoes, M. R.</dc:creator>
<dc:creator>Abe, I.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Sarker, G.</dc:creator>
<dc:creator>Ciccarelli, A.</dc:creator>
<dc:creator>Zhou, L.</dc:creator>
<dc:creator>Lee, C.</dc:creator>
<dc:creator>Martinez-Sanchez, N.</dc:creator>
<dc:creator>Dustin, M.</dc:creator>
<dc:creator>Anderson, K.</dc:creator>
<dc:creator>Zhan, C.</dc:creator>
<dc:creator>Horvath, T.</dc:creator>
<dc:creator>Velloso, L.</dc:creator>
<dc:creator>Kajimura, S.</dc:creator>
<dc:creator>Domingos, A. I.</dc:creator>
<dc:date>2024-05-21</dc:date>
<dc:identifier>doi:10.1101/2024.05.18.594804</dc:identifier>
<dc:title><![CDATA[Sympathetic neuron-derived NPY protects from obesity by sustaining the mural progenitors of thermogenic adipocytes.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.26.595746v1?rss=1">
<title>
<![CDATA[
Marsupial single-cell transcriptomics provides an atlas of developmental heterochrony 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.26.595746v1?rss=1"
</link>
<description><![CDATA[
Single-cell transcriptomics has revealed conserved and divergent programmes of organogenesis in mammals, but existing studies have focused on eutherians. Marsupials exhibit short gestation and complete development externally, necessitating accelerated differentiation of anterior features required for locomotion and feeding. As such, they represent a unique outgroup with which to understand temporal shifts in development, known as heterochrony. Here, we generate the first single-cell transcriptomic atlas of gastrulation and early organogenesis in a marsupial, the opossum Monodelphis domestica. We find that anterior prioritisation is achieved by earlier initiation and shorter duration of transcriptional programmes relative to eutherians. The result is uncoupling of transcriptional and morphological progression, revealing unforeseen diversity in the order of developmental sequences in mammals. We uncover novel tissues for which heterochrony has not previously been documented. Our findings indicate that accelerated activation of transcriptional programmes facilitates the rapid growth needed for survival of the marsupial neonate.
]]></description>
<dc:creator>Menchero, S.</dc:creator>
<dc:creator>Barrington, C.</dc:creator>
<dc:creator>Alanis-Lobato, G.</dc:creator>
<dc:creator>Varsally, W.</dc:creator>
<dc:creator>Niakan, K. K.</dc:creator>
<dc:creator>Turner, J. M. A.</dc:creator>
<dc:date>2024-05-26</dc:date>
<dc:identifier>doi:10.1101/2024.05.26.595746</dc:identifier>
<dc:title><![CDATA[Marsupial single-cell transcriptomics provides an atlas of developmental heterochrony]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.22.595296v1?rss=1">
<title>
<![CDATA[
Genesis: A Modular Protein Language Modelling Approach to Immunogenicity Prediction 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.22.595296v1?rss=1"
</link>
<description><![CDATA[
Neoantigen immunogenicity prediction is a highly challenging problem in the development of personalised medicines. Low reactivity rates in called neoantigens result in a difficult prediction scenario with limited training datasets. Here we describe Genesis, a modular protein language modelling approach to immunogenicity prediction for CD8+ reactive epitopes. Genesis comprises of a pMHC encoding module trained on three pMHC prediction tasks, an optional TCR encoding module and a set of context specific immunogenicity prediction head modules. Compared with state-of-the-art models for each task, Genesis encoding module performs comparably or better on pMHC binding affinity, eluted ligand prediction and stability tasks. Genesis outperforms all compared models on pMHC immunogenicity prediction (Area under the receiver operating characteristic curve=0.619, average precision: 0.514), with a 7% increase in average precision compared to the next best model. Genesis shows further improved performance on immunogenicity prediction with the integration of TCR context information. Genesis performance is further analysed for interpretability, which locates areas of weakness found across existing immunogenicity models and highlight possible biases in public datasets.
]]></description>
<dc:creator>O'Brien, H.</dc:creator>
<dc:creator>Salm, M.</dc:creator>
<dc:creator>Morton, L. T.</dc:creator>
<dc:creator>Szukszto, M.</dc:creator>
<dc:creator>O'Farrell, F.</dc:creator>
<dc:creator>Boulton, C.</dc:creator>
<dc:creator>King, L.</dc:creator>
<dc:creator>Bola, S. K.</dc:creator>
<dc:creator>Becker, P.</dc:creator>
<dc:creator>Craig, A.</dc:creator>
<dc:creator>Nielsen, M.</dc:creator>
<dc:creator>Samuels, Y.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Mansour, M. R.</dc:creator>
<dc:creator>Hadrup, S. R.</dc:creator>
<dc:creator>Quezada, S.</dc:creator>
<dc:date>2024-05-26</dc:date>
<dc:identifier>doi:10.1101/2024.05.22.595296</dc:identifier>
<dc:title><![CDATA[Genesis: A Modular Protein Language Modelling Approach to Immunogenicity Prediction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.27.596037v1?rss=1">
<title>
<![CDATA[
The Role of Metabolism in Shaping Enzyme Structures Over 400 Million Years of Evolution 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.27.596037v1?rss=1"
</link>
<description><![CDATA[
The functions of cells and proteins depend on their biochemical microenvironment. To understand how biochemical constraints shaped protein structural evolution, we coupled the extensive genetic and metabolic data from the Saccharomycotina subphylum with the capability of AlphaFold2 to systematically predict protein structures from sequence. Determining how 11,269 enzyme structures catalysing 361 different metabolic reactions evolved over 400 million years alongside their molecular functions, we report that metabolism has shaped the structural evolution of enzymes at different levels: the organisms overall metabolism; the topological organisation of the metabolic network; and each enzymes molecular properties. For example, structural evolution depends on each enzymes reaction mechanism, on the variability rather than the amount of metabolic flux, and on biosynthetic cost. Evolutionary cost-optimization is stronger on highly abundant enzymes and acts differently on different structural domains, with the exception of small-molecule binding sites, which are prioritised over other structural domains and lack cost-optimisation. Finally, while enzyme surfaces are less constrained, surface residues can also be exposed to positive selection for the co-evolution of protein-protein interaction sites. Accessing AlphaFolds power to predict protein structures systematically and across species barriers, facilitating the integration of protein structures with functional genomics, we were thus able to map biological constraints which shape protein structural evolution at scale and over long timelines.
]]></description>
<dc:creator>Lemke, O.</dc:creator>
<dc:creator>Heineike, B. M.</dc:creator>
<dc:creator>Viknander, S.</dc:creator>
<dc:creator>Cohen, N.</dc:creator>
<dc:creator>Steenwyk, J. L.</dc:creator>
<dc:creator>Spranger, L.</dc:creator>
<dc:creator>Li, F.</dc:creator>
<dc:creator>Agostini, F.</dc:creator>
<dc:creator>Lee, C. T.</dc:creator>
<dc:creator>Aulakh, S. K.</dc:creator>
<dc:creator>Nielsen, J.</dc:creator>
<dc:creator>Rokas, A.</dc:creator>
<dc:creator>Berman, J.</dc:creator>
<dc:creator>Zelezniak, A.</dc:creator>
<dc:creator>Gossmann, T. I.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:date>2024-05-27</dc:date>
<dc:identifier>doi:10.1101/2024.05.27.596037</dc:identifier>
<dc:title><![CDATA[The Role of Metabolism in Shaping Enzyme Structures Over 400 Million Years of Evolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.24.592913v1?rss=1">
<title>
<![CDATA[
The CSF transcriptome in pneumococcal meningitis reveals compartmentalised host inflammatory responses associated with mortality. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.24.592913v1?rss=1"
</link>
<description><![CDATA[
AbstractO_ST_ABSBackgroundC_ST_ABSPneumococcal meningitis (PM) has persistently poor clinical outcomes, especially in sub- Saharan Africa. To better characterise the inflammatory response and identify factors associated with mortality we compared paired peripheral blood and cerebrospinal fluid (CSF) transcriptomes before the initiation of antibiotics in Malawian adults with proven PM.

ResultsBlood transcriptional profiles were obtained in 28 patients with PM, with simultaneous paired with CSF profiles available for 13 patients. 15/28 (52%) patients died. Comparison of the transcriptome between CSF and blood compartments showed upregulation of 2293 differentially expressed genes in CSF and 909 in blood; enriched pathways in CSF included inflammasome activity and neutrophil migration/activation in the CSF, contrasting with enrichment for pathways including platelet and endothelial activation, cell cycle, cytokine release and oxidative stress in the blood transcriptome. Comparison of CSF profiles between survivors and non-survivors revealed 1829 differentially expressed genes, non- survivor CSF was enriched for multiple innate inflammatory pathways, including IL-17A and Type 1 interferons and proteolysis. In contrast, minimal transcriptomic differences between outcome groups were detected in blood.

ConclusionInflammation in PM is characterised by compartmentalised responses in blood and CSF. Poorer outcomes are associated with an dysregulated innate immune host response to S. pneumoniae in the CSF compartment.
]]></description>
<dc:creator>Guerra-Assuncao, J.-A.</dc:creator>
<dc:creator>Chakroborty, P.</dc:creator>
<dc:creator>Pollara, G.</dc:creator>
<dc:creator>Venturini, C.</dc:creator>
<dc:creator>Mlozowa, V. S.</dc:creator>
<dc:creator>Denis, B.</dc:creator>
<dc:creator>Nyirenda, M.</dc:creator>
<dc:creator>Lalloo, D. G.</dc:creator>
<dc:creator>Noursadeghi, M.</dc:creator>
<dc:creator>Heyderman, R. S.</dc:creator>
<dc:creator>Brown, J. S.</dc:creator>
<dc:creator>Wall, E. C.</dc:creator>
<dc:date>2024-05-28</dc:date>
<dc:identifier>doi:10.1101/2024.05.24.592913</dc:identifier>
<dc:title><![CDATA[The CSF transcriptome in pneumococcal meningitis reveals compartmentalised host inflammatory responses associated with mortality.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.24.595769v1?rss=1">
<title>
<![CDATA[
North Pontic crossroads: Mobility in Ukraine from the Bronze Age to the early modern period 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.24.595769v1?rss=1"
</link>
<description><![CDATA[
The North Pontic region, which encompasses present-day Ukraine, was a crossroads of migration as it connected the vast Eurasian Steppe with Central Europe. We generated shotgun-sequenced genomic data for 91 individuals dating from around 7,000 BCE to 1,800 CE to study migration and mobility history in the region, with a particular focus on historically attested migrating groups during the Iron Age and the medieval period, such as Scythian, Chernyakhiv, Saltiv and Nogai associated peoples. We infer a high degree of temporal heterogeneity in ancestry, with fluctuating genetic affinities to present-day Western European, Eastern European, Western Steppe and East Asian groups. We also infer high heterogeneity in ancestry within geographically, culturally and socially defined groups. Despite this, we find that ancestry components which are widespread in Eastern and Central Europe have been present in the Ukraine region since the Bronze Age.
]]></description>
<dc:creator>Saag, L.</dc:creator>
<dc:creator>Utevska, O.</dc:creator>
<dc:creator>Zadnikov, S.</dc:creator>
<dc:creator>Shramko, I.</dc:creator>
<dc:creator>Gorbenko, K.</dc:creator>
<dc:creator>Bandrivskyi, M.</dc:creator>
<dc:creator>Pavliv, D.</dc:creator>
<dc:creator>Bruyako, I.</dc:creator>
<dc:creator>Grechko, D.</dc:creator>
<dc:creator>Okatenko, V.</dc:creator>
<dc:creator>Toshev, G.</dc:creator>
<dc:creator>Andrukh, S.</dc:creator>
<dc:creator>Radziyevska, V.</dc:creator>
<dc:creator>Buynov, Y.</dc:creator>
<dc:creator>Kotenko, V.</dc:creator>
<dc:creator>Smyrnov, O.</dc:creator>
<dc:creator>Petrauskas, O.</dc:creator>
<dc:creator>Magomedov, B.</dc:creator>
<dc:creator>Didenko, S.</dc:creator>
<dc:creator>Heiko, A.</dc:creator>
<dc:creator>Reida, R.</dc:creator>
<dc:creator>Sapiehin, S.</dc:creator>
<dc:creator>Aksonov, V.</dc:creator>
<dc:creator>Laptiev, O.</dc:creator>
<dc:creator>Terskyi, S.</dc:creator>
<dc:creator>Skorokhod, V.</dc:creator>
<dc:creator>Zhyhola, V.</dc:creator>
<dc:creator>Sytyi, Y.</dc:creator>
<dc:creator>Jarve, M.</dc:creator>
<dc:creator>Scheib, C. L.</dc:creator>
<dc:creator>Anastasiadou, K.</dc:creator>
<dc:creator>Kelly, M.</dc:creator>
<dc:creator>Williams, M.</dc:creator>
<dc:creator>Silva, M.</dc:creator>
<dc:creator>Barrington, C.</dc:creator>
<dc:creator>Gilardet, A.</dc:creator>
<dc:creator>Macleod, R.</dc:creator>
<dc:creator>Skoglund, P.</dc:creator>
<dc:creator>Thomas, M. G.</dc:creator>
<dc:date>2024-05-28</dc:date>
<dc:identifier>doi:10.1101/2024.05.24.595769</dc:identifier>
<dc:title><![CDATA[North Pontic crossroads: Mobility in Ukraine from the Bronze Age to the early modern period]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.30.596437v1?rss=1">
<title>
<![CDATA[
The diversity of SNCA transcripts in neurons, and its impact on antisense oligonucleotide therapeutics 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.30.596437v1?rss=1"
</link>
<description><![CDATA[
The role of the SNCA gene locus in driving Parkinsons disease (PD) through rare and common genetic variation is well-recognized, but the transcriptional diversity of SNCA in vulnerable cell types remains unclear. We performed SNCA long-read RNA sequencing in human dopaminergic neurons and show that annotated SNCA transcripts account for only 5% of expression. Rather, the majority of expression (75%) at the SNCA locus originates from transcripts with alternative 5 and 3 untranslated regions. Importantly, 10% originates from transcripts encoding open reading frames not previously annotated, which are translated and detectable in human postmortem brain. Defining the 3 untranslated regions enabled the rational design of antisense oligonucleotides targeting the majority of SNCA transcripts, leading to the effective reversal of PD pathology, including protein aggregation, mitochondrial dysfunction, and toxicity. Resolving the complexity of the SNCA transcriptional landscape impacts RNA therapies and highlights differences in protein isoforms and their contribution to disease.
]]></description>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>Gustavsson, E. K.</dc:creator>
<dc:creator>Doykov, I.</dc:creator>
<dc:creator>Murphy, D.</dc:creator>
<dc:creator>Virdi, G.</dc:creator>
<dc:creator>Lachica, J.</dc:creator>
<dc:creator>Rontgen, A.</dc:creator>
<dc:creator>Murtada, M. H.</dc:creator>
<dc:creator>Pang, C. W.</dc:creator>
<dc:creator>Macpherson, H.</dc:creator>
<dc:creator>Wernick, A. I.</dc:creator>
<dc:creator>Toomey, C. E.</dc:creator>
<dc:creator>Athauda, D.</dc:creator>
<dc:creator>Choi, M.</dc:creator>
<dc:creator>Hardy, J.</dc:creator>
<dc:creator>Wood, N. W.</dc:creator>
<dc:creator>Vendruscolo, M.</dc:creator>
<dc:creator>Mills, K.</dc:creator>
<dc:creator>Heywood, W.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:date>2024-05-31</dc:date>
<dc:identifier>doi:10.1101/2024.05.30.596437</dc:identifier>
<dc:title><![CDATA[The diversity of SNCA transcripts in neurons, and its impact on antisense oligonucleotide therapeutics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.04.597345v1?rss=1">
<title>
<![CDATA[
Growth-induced physiological hypoxia correlates with growth deceleration during normal development 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.04.597345v1?rss=1"
</link>
<description><![CDATA[
Growth deceleration is a universal feature of growth during development: most organs and tissues slow down their growth rate much before growth termination. Using transcriptomics analysis, we show that during their two-day period of growth deceleration, wing imaginal discs of Drosophila undergo a progressive metabolic shift away from oxidative phosphorylation and towards glycolysis. We then develop an ultra-sensitive reporter HIF-1 activity, which reveals that imaginal discs become increasingly hypoxic during development in normoxic conditions, suggesting that limiting oxygen supply could underlie growth deceleration. Growth is energetically expensive and thus expected to contribute, indirectly, to oxygen consumption. Indeed, excess TOR signalling, a key stimulator of growth, triggers hypoxia locally and systemically, highlighting the need to rein in growth when oxygen becomes limiting. This is achieved by a negative feedback loop whereby the classic TOR-inhibitory function of HIF-1 is deployed in response to developmental hypoxia. The absence of Sima/HIF-1 leads to cellular stress, which is alleviated by reduced TOR signalling. Conversely, a small increase in oxygen supply reduces the stress induced by excess TOR activity. We conclude that mild hypoxia is a normal feature of organ development and that Sima/HIF-1 prevents growth-induced oxygen demand from exceeding supply.
]]></description>
<dc:creator>Zhao, Y.</dc:creator>
<dc:creator>Alexandre, C.</dc:creator>
<dc:creator>Kelly, G.</dc:creator>
<dc:creator>Perez-Mockus, G.</dc:creator>
<dc:creator>Vincent, J.-P.</dc:creator>
<dc:date>2024-06-05</dc:date>
<dc:identifier>doi:10.1101/2024.06.04.597345</dc:identifier>
<dc:title><![CDATA[Growth-induced physiological hypoxia correlates with growth deceleration during normal development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.11.598403v1?rss=1">
<title>
<![CDATA[
Illuminating morphogen and patterning dynamics with optogenetic control of morphogen production 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.11.598403v1?rss=1"
</link>
<description><![CDATA[
Cells use dynamic spatial and temporal cues to instruct cell fate decisions during development. Morphogens are key examples, where the concentration and duration of morphogen exposure produce distinct cell fates that drive tissue patterning. Studying the dynamics of these processes has been challenging. Here, we establish an optogenetic system for morphogen production that enables the investigation of developmental patterning in vitro. Using a tunable light-inducible gene expression system, we generate long-range Shh gradients that pattern neural progenitors into spatially distinct progenitor domains mimicking the spatial arrangement of neural progenitors found in vivo during vertebrate neural tube development. With this system, we investigate how biochemical features of Shh and the presence of morphogen-interacting proteins affect the patterning length scale. We measure tissue clearance rates, revealing that Shh has an extracellular half-life of about 1h, and we probe how the level and duration of morphogen exposure govern the acquisition and maintenance of cell fates. The rate of Shh turnover is substantially faster than the downstream gene expression dynamics, indicating that the gradient is continually renewed during patterning. Together the optogenetic approach establishes a simple experimental system for the quantitative interrogation of morphogen patterning. Controlling morphogen dynamics in a reproducible manner provides a framework to dissect the interplay between biochemical cues, the biophysics of gradient formation, and the transcriptional programmes underlying developmental patterning.
]]></description>
<dc:creator>Benzinger, D.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:date>2024-06-11</dc:date>
<dc:identifier>doi:10.1101/2024.06.11.598403</dc:identifier>
<dc:title><![CDATA[Illuminating morphogen and patterning dynamics with optogenetic control of morphogen production]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.10.598250v1?rss=1">
<title>
<![CDATA[
Structural Insight into the Function of Human Peptidyl Arginine Deiminase 6 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.10.598250v1?rss=1"
</link>
<description><![CDATA[
Peptidyl arginine deiminase 6 (PADI6) is vital for early embryonic development in mice and humans, yet its function remains elusive. PADI6 is less conserved than other PADIs and it is currently unknown whether it has a catalytic function. Here we have shown that human PADI6 dimerises like hPADIs 2-4, however, does not bind Ca2+ and is inactive in in vitro assays against standard PADI substrates. By determining the crystal structure of hPADI6, we show that hPADI6 is structured in the absence of Ca2+ where hPADI2 and hPADI4 are not, and the Ca-binding sites are not conserved. Moreover, we show that whilst the key catalytic aspartic acid and histidine residues are structurally conserved, the cysteine is displaced far from the active site centre and the hPADI6 active site pocket appears closed through a unique evolved mechanism in hPADI6, not present in the other PADIs. Taken together, these findings provide insight into how the function of hPADI6 may differ from the other PADIs based on its structure and provides a resource for characterising the damaging effect of clinically significant PADI6 variants.
]]></description>
<dc:creator>Williams, J. P.</dc:creator>
<dc:creator>Mouilleron, S.</dc:creator>
<dc:creator>Hernandez Trapero, R.</dc:creator>
<dc:creator>Bertran, M. T.</dc:creator>
<dc:creator>Marsh, J. A.</dc:creator>
<dc:creator>Walport, L. J.</dc:creator>
<dc:date>2024-06-10</dc:date>
<dc:identifier>doi:10.1101/2024.06.10.598250</dc:identifier>
<dc:title><![CDATA[Structural Insight into the Function of Human Peptidyl Arginine Deiminase 6]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.14.599018v1?rss=1">
<title>
<![CDATA[
Mycobacterium tuberculosis ESX-T7SS impacts the 3D architecture of tuberculous lesion in susceptible mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.14.599018v1?rss=1"
</link>
<description><![CDATA[
Tuberculosis (TB) is characterized by the formation of heterogenous, immune-rich granulomas present in various forms in the lungs. Both host and pathogen contribute to this heterogeneity however the molecular and cellular drivers of within-host granuloma heterogeneity remain to be fully elucidated. This knowledge gap is due to a lack of experimental approaches that can fully capture the complex dynamics of the lung architecture, dynamics of host-pathogen interplay and pathogenesis. Here, we developed an approach that combines PACT-based clearing with light sheet fluorescent microscopy for visualizing lesion architecture development and lung involvement in M. tuberculosis-infected C3HeB/FeJ susceptible mice. This 3D modelling of whole lung lobes approach revealed critical architectural features in lesion development and lung involvement that were not apparent using traditional thin section imaging. Wild type M. tuberculosis infection triggered a clear and well-organized granulomatous-like organization with substantial growth throughout the infection period that invaded a high percentage of the total lung volume. In contrast, infection with the avirulent ESX-1 deletion mutant strain Mtb {Delta}RD1 showed an altered growth pattern with diffuse and sparsely organized CD11b recruitment to sites of infection. Moreover, most of the Mtb {Delta}RD1-triggered lesions were present in the periphery of the lungs and did not display any formal organization. We further provide a novel way of interrogating PACT-cleared tissue for high ultrastructural content using volumetric correlative light and electron microscopy, allowing individual immune cell populations to be tracked and their fate within the granuloma captured. Ultimately, the combination of both modalities allowed an unprecedented view of the architectural distribution of M. tuberculosis in the lungs and the progression of lesion development over time. Our data highlight that ESX-1 from M. tuberculosis is required for lesion architecture progression in a susceptible mouse model of TB.
]]></description>
<dc:creator>Beltran, C. G. G.</dc:creator>
<dc:creator>Kriel, J.</dc:creator>
<dc:creator>Nolan, M. B.</dc:creator>
<dc:creator>Botha, S. M.</dc:creator>
<dc:creator>Ciccarelli, A.</dc:creator>
<dc:creator>Loos, B.</dc:creator>
<dc:creator>Gutierrez, M.</dc:creator>
<dc:creator>Walzl, G.</dc:creator>
<dc:date>2024-06-14</dc:date>
<dc:identifier>doi:10.1101/2024.06.14.599018</dc:identifier>
<dc:title><![CDATA[Mycobacterium tuberculosis ESX-T7SS impacts the 3D architecture of tuberculous lesion in susceptible mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.11.598565v1?rss=1">
<title>
<![CDATA[
Metabolic remodelling in hiPSC-derived myofibres carrying the m.3243A>G mutation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.11.598565v1?rss=1"
</link>
<description><![CDATA[
Mutations in mitochondrial DNA cause severe multisystem disease, frequently associated with muscle weakness. The m.3243A>G mutation is the major cause of Mitochondrial Encephalomyopathy Lactic Acidosis and Stroke Like episodes (MELAS). Experimental models that recapitulate the disease phenotype in vitro for disease modelling or drug screening are very limited. We have therefore generated hiPSC-derived muscle fibres with variable heteroplasmic mtDNA mutation load without significantly affecting muscle differentiation potential. The cells are excitable and show physiological characteristics of muscle fibres and show well organised myofibrillar structure. In cells carrying the m.3243A>G, the mitochondrial membrane potential and oxygen consumption were reduced in relation to the mutant load. We have shown through proteomic, phosphoproteomic, and metabolomic analyses that the m.3243A>G mutation variably affects the cell phenotype in relation to the mutant load. This variation is reflected by an increase in the NADH/NAD+ ratio, which in turns influences key nutrient-sensing pathways in the myofibres. This model enables detailed study of the impact of the mutation on cellular bioenergetics and on muscle physiology with the potential to provide a platform for drug screening.
]]></description>
<dc:creator>Valdebenito, G. E.</dc:creator>
<dc:creator>Chacko, A. R.</dc:creator>
<dc:creator>Chung, C.-Y.</dc:creator>
<dc:creator>Sheshadri, P.</dc:creator>
<dc:creator>Chi, H.</dc:creator>
<dc:creator>O'Callaghan, B.</dc:creator>
<dc:creator>Houlden, H.</dc:creator>
<dc:creator>Rouse, H.</dc:creator>
<dc:creator>Morales, V.</dc:creator>
<dc:creator>Bianchi, K.</dc:creator>
<dc:creator>Tedesco, F. S.</dc:creator>
<dc:creator>Pitceathly, R. D.</dc:creator>
<dc:creator>Duchen, M. R.</dc:creator>
<dc:date>2024-06-13</dc:date>
<dc:identifier>doi:10.1101/2024.06.11.598565</dc:identifier>
<dc:title><![CDATA[Metabolic remodelling in hiPSC-derived myofibres carrying the m.3243A>G mutation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.15.599140v1?rss=1">
<title>
<![CDATA[
Network nature of ligand-receptor interactions underlies disease comorbidity in the brain 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.15.599140v1?rss=1"
</link>
<description><![CDATA[
Neurodegenerative disorders have overlapping symptoms and have high comorbidity rates, but this is not reflected in overlaps of risk genes. We have investigated whether ligand-receptor interactions (LRIs) are a mechanism by which distinct genes associated with disease risk can impact overlapping outcomes. We found that LRIs are likely disrupted in neurological disease and that the ligand-receptor networks associated with neurological diseases have substantial overlaps. Specifically, 96.8% of LRIs associated with disease risk are interconnected in a single LR network. These ligands and receptors are enriched for roles in inflammatory pathways and highlight the role of glia in cross-disease risk. Disruption to this LR network due to disease-associated processes (e.g. differential transcript use, protein misfolding) is likely to contribute to disease progression and risk of comorbidity. Our findings have implications for drug development, as they highlight the potential benefits and risks of pursuing cross-disease drug targets.
]]></description>
<dc:creator>Grant-Peters, M.</dc:creator>
<dc:creator>Fairbrother-Browne, A.</dc:creator>
<dc:creator>Hicks, A.</dc:creator>
<dc:creator>Guo, B.</dc:creator>
<dc:creator>Reynolds, R. H.</dc:creator>
<dc:creator>Huuki-Myers, L. A.</dc:creator>
<dc:creator>Eagles, N.</dc:creator>
<dc:creator>Brenton, J.</dc:creator>
<dc:creator>Garcia-Ruiz, S.</dc:creator>
<dc:creator>Wood, N.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Martinowich, K.</dc:creator>
<dc:creator>Maynard, K.</dc:creator>
<dc:creator>Collado-Torres, L.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:date>2024-06-16</dc:date>
<dc:identifier>doi:10.1101/2024.06.15.599140</dc:identifier>
<dc:title><![CDATA[Network nature of ligand-receptor interactions underlies disease comorbidity in the brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.04.23.590858v1?rss=1">
<title>
<![CDATA[
Inferring gene regulatory networks using DNA methylation data 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.04.23.590858v1?rss=1"
</link>
<description><![CDATA[
We show improved accuracy in-silico of inference of gene-regulatory network (GRN) structure, resulting from the use of an epigenomic prior network. We demonstrate important use-cases of our proposed methodology by re-analysing 12 datasets from 12 different studies. These include data from cells from human embryos, healthy adult tissue, and cancer, and include single-cell mRNA sequencing data, DNA methylation (DNAme) data, chromatin accessibility data, and histone modification data. We find that DNAme data are very effective for inferring the epigenomic prior network, recapitulating known epigenomic network structure found previously from chromatin accessibility data. Furthermore, we find that inferring the epigenomic prior network from DNAme data reveals candidate TF cis-regulations for at least eight times as many genes, when compared with chromatin accessibility data. When our proposed methodology is applied to real datasets from human embryonic development and from women at risk of breast cancer, we find patterns of differential cis-regulation that are in line with expectations under appropriate biological models, and that may be used to propose hypotheses about pre-cancerous epigenomic changes.
]]></description>
<dc:creator>Bartlett, T. E.</dc:creator>
<dc:date>2024-04-25</dc:date>
<dc:identifier>doi:10.1101/2024.04.23.590858</dc:identifier>
<dc:title><![CDATA[Inferring gene regulatory networks using DNA methylation data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.24.600474v1?rss=1">
<title>
<![CDATA[
SubtiToolKit - a bioengineering kit for Bacillus subtilis and Gram-positive bacteria. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.24.600474v1?rss=1"
</link>
<description><![CDATA[
Building DNA constructs of increasing complexity is key to synthetic biology. Golden Gate methods led to the creation of cloning toolkits - collections of modular standardized DNA parts hosted on hierarchic plasmids, developed for yeast, plants, Gram-negative bacteria, and human cells. However, Gram-positive bacteria have been neglected. Bacillus subtilis is a Gram-positive model organism and a workhorse in the bioindustry. Here, we present the SubtiToolKit, a high- efficiency cloning toolkit for B. subtilis and Gram-positive bacteria. Its design permits DNA constructs for transcriptional units, operons, knock-in and knock-out applications. It contains libraries of promoters, RBSs, fluorescent proteins, protein tags, terminators, genome integration parts, a no-leakage genetic device to control the expression of toxic products during E. coli assembly, and a toolbox for industrially relevant strains of Geobacillus and Parageobacillus as an example of SubtiToolKit versatility for other Gram-positive bacteria and its future perspective as a reference toolkit.
]]></description>
<dc:creator>Caro Astorga, J.</dc:creator>
<dc:creator>Malci, K.</dc:creator>
<dc:creator>Ellis, T.</dc:creator>
<dc:creator>James, P.</dc:creator>
<dc:creator>Debenedictis, E.</dc:creator>
<dc:creator>Ming, H.</dc:creator>
<dc:creator>Rogan, M.</dc:creator>
<dc:date>2024-06-25</dc:date>
<dc:identifier>doi:10.1101/2024.06.24.600474</dc:identifier>
<dc:title><![CDATA[SubtiToolKit - a bioengineering kit for Bacillus subtilis and Gram-positive bacteria.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.01.601514v1?rss=1">
<title>
<![CDATA[
Dynamics of positional information in the vertebrate neural tube 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.01.601514v1?rss=1"
</link>
<description><![CDATA[
In developing embryos, cells acquire distinct identities depending on their position in a tissue. Secreted signaling molecules, known as morphogens, act as long-range cues to provide the spatial information that controls these cell fate decisions. In several tissues, both the level and the duration of morphogen signaling appear to be important for determining cell fates. This is the case in the forming vertebrate nervous system where antiparallel morphogen gradients pattern the dorsal-ventral axis by partitioning the tissue into sharply delineated domains of molecularly distinct neural progenitors. How information in the gradients is decoded to generate precisely positioned boundaries of gene expression remains an open question. Here, we adopt tools from information theory to quantify the positional information in the neural tube and investigate how temporal changes in signaling could influence positional precision. The results reveal that the use of signaling dynamics, as well as the signaling level, substantially increases the precision possible for the estimation of position from morphogen gradients. This analysis links the dynamics of opposing morphogen gradients with precise pattern formation and provides an explanation for why time is used to impart positional information.
]]></description>
<dc:creator>Markovic, A.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Page, K. M.</dc:creator>
<dc:date>2024-07-01</dc:date>
<dc:identifier>doi:10.1101/2024.07.01.601514</dc:identifier>
<dc:title><![CDATA[Dynamics of positional information in the vertebrate neural tube]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.03.601939v1?rss=1">
<title>
<![CDATA[
Pan-cancer subclonal mutation analysis of 7,827 tumors predicts clinical outcome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.03.601939v1?rss=1"
</link>
<description><![CDATA[
Intra-tumor heterogeneity is characterized by a diverse population of tumor clones and subclones which are important drivers of tumor evolution and therapeutic response. However, accurate subclonal reconstruction at scale remains challenging. We developed a machine learning tool, CliPP, and surveyed 9,972 tumors from 32 cancer types. We found that high subclonal mutation load (sML), the fraction of subclonal single nucleotide variants (SNVs) to all SNVs in the coding region, was prognostic of survival (progression free survival or overall survival) in 18 cancer types. In 14 cancers with low to moderate tumor mutation burden (TMB), high sML was associated with better prognosis. In immunotherapy trials for 42 metastatic prostate cancer (mCRPC), high sML was predictive of favorable response to ipilimumab and associated with increased CD8+ T-cell infiltration and decreased macrophage population. A validation using 613 whole-genomes of esophageal adenocarcinoma confirms the favorable effect of high sML and the observed tumor-associated macrophage. Our study identifies sML as a key feature of cancer, suggesting a biphasic relationship between evolutionary dynamics and differential immune environments. Finally, sML may serve as an orthogonal approach to identify likely responders of immune checkpoint blockade in low to moderate TMB tumors.
]]></description>
<dc:creator>Jiang, Y.</dc:creator>
<dc:creator>Montierth, M. D.</dc:creator>
<dc:creator>Yu, K.</dc:creator>
<dc:creator>Ji, S.</dc:creator>
<dc:creator>Guo, S.</dc:creator>
<dc:creator>Tran, Q.</dc:creator>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Shin, S. J.</dc:creator>
<dc:creator>Cao, S.</dc:creator>
<dc:creator>Li, R.</dc:creator>
<dc:creator>Tang, Y.</dc:creator>
<dc:creator>Lesluyes, T.</dc:creator>
<dc:creator>Kopetz, S.</dc:creator>
<dc:creator>Ajani, J.</dc:creator>
<dc:creator>Msaouel, P.</dc:creator>
<dc:creator>Subudhi, S. K.</dc:creator>
<dc:creator>Aparicio, A.</dc:creator>
<dc:creator>Sharma, P.</dc:creator>
<dc:creator>Shen, J. P.</dc:creator>
<dc:creator>Sood, A. K.</dc:creator>
<dc:creator>Tarabichi, M.</dc:creator>
<dc:creator>Wang, J. R.</dc:creator>
<dc:creator>Kimmel, M.</dc:creator>
<dc:creator>Loo, P. V.</dc:creator>
<dc:creator>Zhu, H.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:date>2024-07-06</dc:date>
<dc:identifier>doi:10.1101/2024.07.03.601939</dc:identifier>
<dc:title><![CDATA[Pan-cancer subclonal mutation analysis of 7,827 tumors predicts clinical outcome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.08.602480v1?rss=1">
<title>
<![CDATA[
Human Gastric Multi-Regional Assembloids Favour Functional Parietal Maturation and Allow Modelling of Antral Foveolar Hyperplasia 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.08.602480v1?rss=1"
</link>
<description><![CDATA[
Patient-derived human organoids have the remarkable capacity to self-organise into more complex structures. However, to what extent gastric organoids can recapitulate human stomach physiological functions remain unexplored. Here, we report how region-specific gastric organoids can self-assemble into complex multi-regional assembloids showing functional response to drugs targeting the ATPase H+/K+ pump. The assembloids show preserved fundus, body, and antrum regional identity, and gastric-specific crosstalk pathways arise. The increased complexity and cross-communication between the different gastric regions, allow for the emergence of the elusive parietal cell type, responsible for the production of gastric acid, with functional response to drugs targeting the ATPase H+/K+ pump. Remarkably, we generated assembloids from PMM2-HIPKD-IBD paediatric patients (Phosphomannomutase 2 - Hyperinsulinemic hypoglycaemia and autosomal recessive polycystic kidney disease - Inflammatory bowel disease), a genetic condition found to be associated with unusual antral foveolar hyperplasia and hyperplastic polyposis. The cellular mechanisms behind such phenomena are poorly understood, and an exhaustive experimental model is needed. The {Delta}PMM2 multi-regional assembloid we have generated efficiently recapitulates hyperplastic-like antral regions, with decreased mucin secretion and glycosylated ATP4b, which results in impaired gastric acid secretion. Multi-regional gastric assembloids, generated using adult-stem cell-derived organoids, successfully recapitulate the structural and functional characteristics of the human stomach, offering a promising tool for studying gastric epithelial interactions and disease mechanisms previously challenging to investigate in primary models.
]]></description>
<dc:creator>Jones, B. C.</dc:creator>
<dc:creator>Benedetti, G.</dc:creator>
<dc:creator>Cala, G.</dc:creator>
<dc:creator>Tullie, L.</dc:creator>
<dc:creator>Simcock, I.</dc:creator>
<dc:creator>Lutman, R.</dc:creator>
<dc:creator>Balys, M.</dc:creator>
<dc:creator>Amiri, R.</dc:creator>
<dc:creator>Sufi, J.</dc:creator>
<dc:creator>Arthurs, O.</dc:creator>
<dc:creator>Eaton, S.</dc:creator>
<dc:creator>Anderson, G.</dc:creator>
<dc:creator>Elvassore, N.</dc:creator>
<dc:creator>Li, V. S.</dc:creator>
<dc:creator>Jones, K. D.</dc:creator>
<dc:creator>Tape, C. J.</dc:creator>
<dc:creator>Luni, C.</dc:creator>
<dc:creator>Giobbe, G. G.</dc:creator>
<dc:creator>De Coppi, P.</dc:creator>
<dc:date>2024-07-11</dc:date>
<dc:identifier>doi:10.1101/2024.07.08.602480</dc:identifier>
<dc:title><![CDATA[Human Gastric Multi-Regional Assembloids Favour Functional Parietal Maturation and Allow Modelling of Antral Foveolar Hyperplasia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.12.603249v1?rss=1">
<title>
<![CDATA[
The bat Influenza A virus subtype H18N11 induces nanoscale MHCII clustering upon host cell attachment 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.12.603249v1?rss=1"
</link>
<description><![CDATA[
Prior to the discovery of the bat influenza A virus (IAV) subtypes H17N10 and H18N11, it was believed that all IAVs bind via the viral hemagglutinin (HA) to sialic acid residues to mediate attachment and subsequent viral entry. Both HA subtypes, H17 and H18, however, do not bind to sialic acids but instead engage a proteinaceous receptor: major histocompatibility complex class II (MHCII). The mechanistic details of this hitherto unknown protein-mediated entry are not understood. Since conventional IAVs require attachment to clusters of sialylated glycans to overcome the low affinity of the HA-sialic acid interaction, we hypothesized that bat HA would likewise interact with multiple MHCII molecules. Here, we used photoactivated localization microscopy (PALM) on fixed and live cells expressing MHCII fused to appropriate fluorescent reporters. We show that bat IAV particles attach to pre-existing MHCII clusters present on susceptible cells and that the local HA-MHCII interaction results in an increased cluster size. To measure the impact of viral attachment on the dynamics of MHCII, we utilized an experimental setup designated "inverse infection" where intact viral particles were immobilized on coverslips before live MHCII-expressing cells were seeded on top. Recording the trajectories of single MHCII molecules, this approach revealed that the mobility of MHCII was indeed slowed down in viral proximity leading to a local enrichment of MHCII molecules beneath the viral particle. Taken together, these data suggest that attachment of viral particles leads to clustering of MHCII, a process similar to the MHCII dynamics observed during the formation of an immunological synapse.
]]></description>
<dc:creator>Osman, M. K.</dc:creator>
<dc:creator>Robert, J.</dc:creator>
<dc:creator>Broich, L.</dc:creator>
<dc:creator>Frank, D.</dc:creator>
<dc:creator>Grosse, R.</dc:creator>
<dc:creator>Schwemmle, M.</dc:creator>
<dc:creator>Wrobel, A.</dc:creator>
<dc:creator>Ciminski, K.</dc:creator>
<dc:creator>Sieben, C.</dc:creator>
<dc:creator>Reuther, P.</dc:creator>
<dc:date>2024-07-12</dc:date>
<dc:identifier>doi:10.1101/2024.07.12.603249</dc:identifier>
<dc:title><![CDATA[The bat Influenza A virus subtype H18N11 induces nanoscale MHCII clustering upon host cell attachment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.12.603302v1?rss=1">
<title>
<![CDATA[
PP1 PNUTS binds the restrictor and dephosphorylates RNA pol II CTD Ser5 to stimulate transcription termination 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.12.603302v1?rss=1"
</link>
<description><![CDATA[
The restrictor, ZC3H4/WDR82, is the major termination factor for antisense transcription from bidirectional promoters, but its mechanism is poorly understood. We report that ZC3H4/WDR82 co-purifies with PP1 phosphatase and PP1 phosphatase nuclear targeting subunit, PNUTS, which binds directly to the WDR82 subunit of restrictor. AlphaFold predicts a quaternary complex, PPWZ, in which PP1-associated PNUTS and ZC3H4 both contact WDR82. To investigate the role of protein dephosphorylation in PPWZ activity, we expressed a substrate trap comprising inactive PP1H66K linked to the PNUTS C-terminus. PP1H66K-PNUTS binds pol II large subunit and nuclear exosome components. PP1H66K-PNUTS, but not PP1WT-PNUTS, functions as a dominant-negative inhibitor of antisense termination and CTD Ser5 dephosphorylation. Both these activities require the PNUTS WDR82 binding domain that interacts with restrictor. We show that CTD Ser5 hyperphosphorylation is associated with higher processivity and reduced pausing that would counteract termination, and propose that Ser5 dephosphorylation by PPWZ is coupled to termination. In summary, we identify the PP1 phosphatase activity of the PPWZ complex as essential for terminator function and propose that this heterotetramer is the physiologically relevant form of restrictor.
]]></description>
<dc:creator>Bentley, D.</dc:creator>
<dc:creator>Treisman, R.</dc:creator>
<dc:creator>Erickson, B.</dc:creator>
<dc:creator>Fong, N.</dc:creator>
<dc:creator>Hansen, K.</dc:creator>
<dc:creator>Sheridan, R. M.</dc:creator>
<dc:creator>Larson, K.</dc:creator>
<dc:creator>Saviola, A.</dc:creator>
<dc:creator>Fedoryshchak, R.</dc:creator>
<dc:creator>Mouilleron, S.</dc:creator>
<dc:date>2024-07-13</dc:date>
<dc:identifier>doi:10.1101/2024.07.12.603302</dc:identifier>
<dc:title><![CDATA[PP1 PNUTS binds the restrictor and dephosphorylates RNA pol II CTD Ser5 to stimulate transcription termination]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.16.603765v1?rss=1">
<title>
<![CDATA[
Molecular basis of FIGNL1 in dissociating RAD51 from DNA and chromatin 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.16.603765v1?rss=1"
</link>
<description><![CDATA[
Maintaining genome integrity is an essential and challenging process. RAD51 recombinase, the central player of several crucial processes in repairing and protecting genome integrity, forms filaments on DNA. RAD51 filaments are tightly regulated. One of these regulators is FIGNL1, that prevents persistent RAD51 foci post-damage and genotoxic chromatin association in cells. The cryogenic electron microscopy structure of FIGNL1 in complex with RAD51 reveals that the FIGNL1 forms a non-planar hexamer and RAD51 N-terminus is enclosed in the FIGNL1 hexamer pore. Mutations in pore loop or catalytic residues of FIGNL1 render it defective in filament disassembly and are lethal in mouse embryonic stem cells. Our study reveals a unique mechanism for removing RAD51 from DNA and provides the molecular basis for FIGNL1 in maintaining genome stability.
]]></description>
<dc:creator>Carver, A.</dc:creator>
<dc:creator>Yu, T.-Y.</dc:creator>
<dc:creator>Yates, L.</dc:creator>
<dc:creator>White, T.</dc:creator>
<dc:creator>Wang, R.</dc:creator>
<dc:creator>Lister, K.</dc:creator>
<dc:creator>Jasin, M.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:date>2024-07-16</dc:date>
<dc:identifier>doi:10.1101/2024.07.16.603765</dc:identifier>
<dc:title><![CDATA[Molecular basis of FIGNL1 in dissociating RAD51 from DNA and chromatin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.18.604111v1?rss=1">
<title>
<![CDATA[
Branched actin networks in innate immune cells mediate host microbiota homeostasis. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.18.604111v1?rss=1"
</link>
<description><![CDATA[
Assembly of branched actin networks, driven by the Arp2/3 complex are essential for the function and integrity of the immune system. Patients with loss-of-function mutations in the ARPC5 subunit of the Arp2/3 complex develop inflammation and immunodeficiency after birth, leading to early mortality. However, the mechanistic basis for these phenotypes remains obscure. Here we demonstrate that loss of Arpc5 in the murine hematopoietic system, but not the corresponding Arpc5l isoform causes early-onset intestinal inflammation after weaning. This condition is initiated by microbiota breaching the ileal mucosa, leading to local and systemic inflammation. Macrophage and neutrophils infiltrate into the ileum, but in the absence of Arpc5 fail to restrict microbial invasion. Loss of Arpc5 compromises the ability of macrophages to phagocytose and kill intra-cellular bacteria. Our results underscore the indispensable role of Arpc5, but not Arpc5l containing Arp2/3 complexes in mononuclear phagocytes function and host-microbiota homeostasis.

One-Sentence SummaryArpc5 containing Arp2/3 complexes are essential for host-microbiota homeostasis
]]></description>
<dc:creator>Vasconcellos, L. R.</dc:creator>
<dc:creator>Huang, S. C. M.</dc:creator>
<dc:creator>Suarez-Bonnet, A.</dc:creator>
<dc:creator>Priestnall, S.</dc:creator>
<dc:creator>Varsani-Brown, S.</dc:creator>
<dc:creator>Winder, M. L.</dc:creator>
<dc:creator>Shah, K.</dc:creator>
<dc:creator>Kogata, N.</dc:creator>
<dc:creator>Stockinger, B.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:date>2024-07-18</dc:date>
<dc:identifier>doi:10.1101/2024.07.18.604111</dc:identifier>
<dc:title><![CDATA[Branched actin networks in innate immune cells mediate host microbiota homeostasis.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.18.603748v1?rss=1">
<title>
<![CDATA[
Ancient Borrelia genomes document the evolutionary history of louse-borne relapsing fever 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.18.603748v1?rss=1"
</link>
<description><![CDATA[
Several disease-causing bacteria have transitioned from tick-borne to louse-borne transmission, a process associated with increased virulence and genome reduction. However, the historical time frame and speed of such evolutionary transitions have not been documented with ancient genomes. Here, we discover four ancient cases of Borrelia recurrentis, the causative agent of louse-borne relapsing fever, in Britain between [~]600 and 2,300 years ago, and sequence whole genomes up to 29-fold coverage. We estimate a recent divergence from the closest tick-borne ancestor, likely within the last [~]8,000 years. We reconstruct a chronology of gene losses and acquisitions using the pan-genome of related species, and show that almost all of the reductive evolution observed in B. recurrentis had occurred by [~]2,000 years ago, and was thus a rapid process after divergence. Our observations provide a new understanding of the origins of B. recurrentis and document complex reductive evolution in a specialist vector-borne pathogen.
]]></description>
<dc:creator>Swali, P.</dc:creator>
<dc:creator>Booth, T.</dc:creator>
<dc:creator>Tan, C. C. S.</dc:creator>
<dc:creator>McCabe, J.</dc:creator>
<dc:creator>Anastasiadou, K.</dc:creator>
<dc:creator>Barrington, C.</dc:creator>
<dc:creator>Borrini, M.</dc:creator>
<dc:creator>Bricking, A.</dc:creator>
<dc:creator>Buckberry, J.</dc:creator>
<dc:creator>Buster, L.</dc:creator>
<dc:creator>Carlin, R.</dc:creator>
<dc:creator>Gilardet, A.</dc:creator>
<dc:creator>Glocke, I.</dc:creator>
<dc:creator>Irish, J.</dc:creator>
<dc:creator>Kelly, M.</dc:creator>
<dc:creator>King, M.</dc:creator>
<dc:creator>Petchey, F.</dc:creator>
<dc:creator>Peto, J.</dc:creator>
<dc:creator>Silva, M.</dc:creator>
<dc:creator>Speidel, L.</dc:creator>
<dc:creator>Tait, F.</dc:creator>
<dc:creator>Teoaca, A.</dc:creator>
<dc:creator>Valoriani, S.</dc:creator>
<dc:creator>Williams, M.</dc:creator>
<dc:creator>Madgwick, R.</dc:creator>
<dc:creator>Mullan, G.</dc:creator>
<dc:creator>Wilson, L.</dc:creator>
<dc:creator>Cootes, K.</dc:creator>
<dc:creator>Armit, I.</dc:creator>
<dc:creator>Gutierrez, M. G.</dc:creator>
<dc:creator>van Dorp, L.</dc:creator>
<dc:creator>Skoglund, P.</dc:creator>
<dc:date>2024-07-19</dc:date>
<dc:identifier>doi:10.1101/2024.07.18.603748</dc:identifier>
<dc:title><![CDATA[Ancient Borrelia genomes document the evolutionary history of louse-borne relapsing fever]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.18.604063v1?rss=1">
<title>
<![CDATA[
Systematic identification of Y-chromosome gene functions in mouse spermatogenesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.18.604063v1?rss=1"
</link>
<description><![CDATA[
The mammalian Y chromosome is essential for male fertility, but how individual Y genes regulate spermatogenesis is poorly understood. To resolve this question, we generate a deletion series of the mouse Y chromosome, creating thirteen Y-deletant mouse models and conducting exhaustive reproductive phenotyping. Eight Y genes, including several that are deeply conserved and exhibit testis-specific expression, are dispensable for spermatogenesis. For others, we uncover novel functions, including a role for Uty in establishment and differentiation of spermatogonia, and for Zfy2 in ensuring meiotic pairing and reciprocal recombination between the sex chromosomes. We also generate the first mouse equivalent of the human infertility AZFa deletion, revealing cumulative detrimental effects of Y-gene loss on spermatogenesis. We use single nuclei RNAseq to identify candidate mechanisms by which Y genes regulate the germ cell transcriptome and reveal an unexpected impact of Y genes on testis supporting cells. Our study represents a paradigm for the complete functional dissection of a mammalian Y chromosome and advances our knowledge of human infertility and Y-chromosome evolution.
]]></description>
<dc:creator>Subrini, J.</dc:creator>
<dc:creator>Varsally, W.</dc:creator>
<dc:creator>Balaguer Balsells, I.</dc:creator>
<dc:creator>Bensberg, M.</dc:creator>
<dc:creator>Sioutas, G.</dc:creator>
<dc:creator>Ojarikre, O.</dc:creator>
<dc:creator>Maciulyte, V.</dc:creator>
<dc:creator>Gylemo, B.</dc:creator>
<dc:creator>Crawley, K.</dc:creator>
<dc:creator>Courtis, K.</dc:creator>
<dc:creator>de Rooij, D. G.</dc:creator>
<dc:creator>Turner, J. M.</dc:creator>
<dc:date>2024-07-19</dc:date>
<dc:identifier>doi:10.1101/2024.07.18.604063</dc:identifier>
<dc:title><![CDATA[Systematic identification of Y-chromosome gene functions in mouse spermatogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.19.604101v1?rss=1">
<title>
<![CDATA[
Structures of TGF-beta with betaglycan and the signaling receptors reveal the mechanism whereby betaglycan potentiates receptor complex assembly and signaling 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.19.604101v1?rss=1"
</link>
<description><![CDATA[
Betaglycan (BG) is a transmembrane co-receptor of the transforming growth factor-{beta} (TGF-{beta}) family of signaling ligands. It is essential for embryonic development and tissue homeostasis and fertility in adults. It functions by enabling binding of the three TGF-{beta} isoforms to their signaling receptors and is additionally required for inhibin A (InhA) activity. Despite its requirement for the functions of TGF-{beta}s and InhA in vivo, structural information explaining BG ligand selectivity and its mechanism of action is lacking. Here, we determine the structure of TGF-{beta} bound both to BG and the signaling receptors, TGFBR1 and TGFBR2. We identify key regions responsible for ligand engagement, which has revealed novel binding interfaces that differ from those described for the closely related co-receptor of the TGF-{beta} family, endoglin, thus demonstrating remarkable evolutionary adaptation to enable ligand selectivity. Finally, we provide a structural explanation for the hand-off mechanism underlying TGF-{beta} signal potentiation.
]]></description>
<dc:creator>Wieteska, L.</dc:creator>
<dc:creator>Taylor, A. B.</dc:creator>
<dc:creator>Punch, E.</dc:creator>
<dc:creator>Coleman, J. A.</dc:creator>
<dc:creator>Conway, I. O.</dc:creator>
<dc:creator>Lin, Y.-F.</dc:creator>
<dc:creator>Byeon, C.-H.</dc:creator>
<dc:creator>Hinck, C. S.</dc:creator>
<dc:creator>Krzysiak, T.</dc:creator>
<dc:creator>Ishima, R.</dc:creator>
<dc:creator>Lopez-Casillas, F.</dc:creator>
<dc:creator>Cherepanov, P.</dc:creator>
<dc:creator>Bernard, D. J.</dc:creator>
<dc:creator>Hill, C. S.</dc:creator>
<dc:creator>Hinck, A. P.</dc:creator>
<dc:date>2024-07-22</dc:date>
<dc:identifier>doi:10.1101/2024.07.19.604101</dc:identifier>
<dc:title><![CDATA[Structures of TGF-beta with betaglycan and the signaling receptors reveal the mechanism whereby betaglycan potentiates receptor complex assembly and signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.18.604051v1?rss=1">
<title>
<![CDATA[
Multi-Omic Analysis Reveals Lipid Dysregulation Associated with Mitochondrial Dysfunction in Parkinson's Disease Brain 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.18.604051v1?rss=1"
</link>
<description><![CDATA[
Parkinsons Disease (PD) is an increasingly prevalent condition within the aging population. PD can be attributed to rare genetic mutations, but most cases are sporadic where the gene-environment interactions are unknown/likely contributory. Age related dysregulation of the glycosphingolipid degradation pathway has been implicated in the development of PD, however, our understanding of how brain lipids vary across different regions of the brain, with age and in disease stages, remains limited.

In this study we profiled several phospho- and sphingolipid classes in eight distinct regions of the human brain and investigated the association of lipids with a spatio-temporal pathology gradient, utilising PD samples from early, mid, and late stages of the disease. We performed high-precision tissue sampling in conjunction with targeted LC-MS/MS and applied this to post-mortem samples from PD and control subjects. The lipids were analysed for correlations with untargeted proteomics and mitochondrial activity data, in a multi-omics approach. We concluded that the different brain regions demonstrated their own distinct profiles and also found that several lipids were correlated with age. The strongest differences between PD and controls were identified in ganglioside, sphingomyelin and n-hexosylceramides. Sphingomyelin was also found to correlate with several proteins implicated in Parkinsons disease pathways. Mitochondrial activity was correlated with the levels of several lipids in the putamen region. Finally, we identified a gradient corresponding to Braaks disease spread across the brain regions, where the areas closer to the brainstem/substantia nigra showed alterations in PC, LPC and glycosphingolipids, while the cortical regions showed changes in glycosphingolipids, specifically gangliosides, HexCer and Hex2Cer.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=116 SRC="FIGDIR/small/604051v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Hallqvist, J.</dc:creator>
<dc:creator>Toomey, C. E.</dc:creator>
<dc:creator>Pinto, R.</dc:creator>
<dc:creator>Wernick, A.</dc:creator>
<dc:creator>Shahrani, M.</dc:creator>
<dc:creator>Heales, S.</dc:creator>
<dc:creator>Eaton, S.</dc:creator>
<dc:creator>Mills, K.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Heywood, W. E.</dc:creator>
<dc:date>2024-07-22</dc:date>
<dc:identifier>doi:10.1101/2024.07.18.604051</dc:identifier>
<dc:title><![CDATA[Multi-Omic Analysis Reveals Lipid Dysregulation Associated with Mitochondrial Dysfunction in Parkinson's Disease Brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.22.604575v1?rss=1">
<title>
<![CDATA[
STK19 facilitates the clearance of lesion-stalled RNAPII during transcription-coupled DNA repair 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.22.604575v1?rss=1"
</link>
<description><![CDATA[
Transcription-coupled DNA repair (TCR) removes bulky DNA lesions impeding RNA polymerase II (RNAPII) transcription. Recent studies have outlined the stepwise assembly of TCR factors CSB, CSA, UVSSA, and TFIIH around lesion-stalled RNAPII. However, the mechanism and factors required for the transition to downstream repair steps, including RNAPII removal to provide repair proteins access to the DNA lesion, remain unclear. Here, we identify STK19 as a new TCR factor facilitating this transition. Loss of STK19 does not impact initial TCR complex assembly or RNAPII ubiquitylation but delays lesion-stalled RNAPII clearance, thereby interfering with the downstream repair reaction. Cryo-EM and mutational analysis reveal that STK19 associates with the TCR complex, positioning itself between RNAPII, UVSSA, and CSA. The structural insights and molecular modeling suggest that STK19 positions the ATPase subunits of TFIIH onto DNA in front of RNAPII. Together, these findings provide new insights into the factors and mechanisms required for TCR.
]]></description>
<dc:creator>van den Heuvel, D.</dc:creator>
<dc:creator>Rodriguez-Martinez, M.</dc:creator>
<dc:creator>van der Meer, P. J.</dc:creator>
<dc:creator>Nieto Moreno, N.</dc:creator>
<dc:creator>Park, J.</dc:creator>
<dc:creator>Kim, H.-S.</dc:creator>
<dc:creator>van Schie, J. J.</dc:creator>
<dc:creator>Wondergem, A. P.</dc:creator>
<dc:creator>D'Souza, A.</dc:creator>
<dc:creator>Yakoub, G.</dc:creator>
<dc:creator>Herlihy, A.</dc:creator>
<dc:creator>Kashyap, K.</dc:creator>
<dc:creator>Boissiere, T.</dc:creator>
<dc:creator>Walker, J.</dc:creator>
<dc:creator>Mitter, R.</dc:creator>
<dc:creator>Apelt, K.</dc:creator>
<dc:creator>de Lint, K.</dc:creator>
<dc:creator>Kirdok, I.</dc:creator>
<dc:creator>Ljungman, M.</dc:creator>
<dc:creator>Wolthuis, R. M.</dc:creator>
<dc:creator>Cramer, P.</dc:creator>
<dc:creator>Scharer, O. D.</dc:creator>
<dc:creator>Kokic, G.</dc:creator>
<dc:creator>Svejstrup, J. Q.</dc:creator>
<dc:creator>Luijsterburg, M. S.</dc:creator>
<dc:date>2024-07-22</dc:date>
<dc:identifier>doi:10.1101/2024.07.22.604575</dc:identifier>
<dc:title><![CDATA[STK19 facilitates the clearance of lesion-stalled RNAPII during transcription-coupled DNA repair]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.22.604574v1?rss=1">
<title>
<![CDATA[
Super-resolution imaging of proteins inside live mammalian cells with mLIVE-PAINT 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.22.604574v1?rss=1"
</link>
<description><![CDATA[
Super-resolution microscopy has revolutionized biological imaging, enabling the visualization of structures at the nanometer length scale. Its application in live cells, however, has remained challenging. To address this, we adapted LIVE-PAINT, an approach we established in yeast, for application in live mammalian cells. Using the 101A/101B coiled-coil peptide pair as a peptide-based targeting system, we successfully demonstrate the super-resolution imaging of two distinct proteins in mammalian cells, one localized in the nucleus, and the second in the cytoplasm. This study highlights the versatility of LIVE-PAINT, suggesting its potential for live-cell super-resolution imaging across a range of protein targets in mammalian cells. We name the mammalian cell version of our original method mLIVE-PAINT.
]]></description>
<dc:creator>Bhaskar, H.</dc:creator>
<dc:creator>Gidden, Z.</dc:creator>
<dc:creator>Virdi, G.</dc:creator>
<dc:creator>Kleinjan, D.-J.</dc:creator>
<dc:creator>Rosser, S. J.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Regan, L.</dc:creator>
<dc:creator>Horrocks, M. H.</dc:creator>
<dc:date>2024-07-22</dc:date>
<dc:identifier>doi:10.1101/2024.07.22.604574</dc:identifier>
<dc:title><![CDATA[Super-resolution imaging of proteins inside live mammalian cells with mLIVE-PAINT]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.23.604321v1?rss=1">
<title>
<![CDATA[
ATG9A and ARFIP2 cooperate to regulate PI4P levels for lysosomal repair 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.23.604321v1?rss=1"
</link>
<description><![CDATA[
Lysosome damage activates multiple pathways to prevent lysosome-dependent cell death, including a repair mechanism involving ER-lysosome membrane contact sites, phosphatidylinositol 4-kinase- 2a (PI4K2A), phosphatidylinositol-4 phosphate (PI4P) and oxysterol-binding protein-related proteins (ORPs), lipid transfer proteins. PI4K2A localizes to trans-Golgi network and endosomes yet how it is delivered to damaged lysosomes remains unknown. During acute sterile damage, and damage caused by intracellular bacteria, we show that ATG9A-containing vesicles perform a critical role in delivering PI4K2A to damaged lysosomes. ADP ribosylation factor interacting protein 2 (ARFIP2), a component of ATG9A vesicles, binds and sequesters PI4P on lysosomes, balancing ORP- dependent lipid transfer and promoting retrieval of ATG9A vesicles through recruitment of the adaptor protein complex-3 (AP-3). Our results reveal a role for mobilized ATG9A vesicles and ARFIP2 in lysosome homeostasis after damage and bacterial infection.
]]></description>
<dc:creator>De Tito, S.</dc:creator>
<dc:creator>Almacellas, E.</dc:creator>
<dc:creator>Yu, D. D.</dc:creator>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Millard, E.</dc:creator>
<dc:creator>Hervas, J. H.</dc:creator>
<dc:creator>Pellegrino, E.</dc:creator>
<dc:creator>Panagi, I.</dc:creator>
<dc:creator>Fodge, D.</dc:creator>
<dc:creator>Thurston, T. L. M.</dc:creator>
<dc:creator>Gutierrez, M.</dc:creator>
<dc:creator>Tooze, S.</dc:creator>
<dc:date>2024-07-24</dc:date>
<dc:identifier>doi:10.1101/2024.07.23.604321</dc:identifier>
<dc:title><![CDATA[ATG9A and ARFIP2 cooperate to regulate PI4P levels for lysosomal repair]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.24.604962v1?rss=1">
<title>
<![CDATA[
Multiscale mechanics drive functional maturation of the vertebrate heart 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.24.604962v1?rss=1"
</link>
<description><![CDATA[
How simple tissue primordia sculpt complex functional organs, robustly and reproducibly, remains elusive. As the zebrafish embryo grows into a larva, to improve its heart function, the embryonic myocardial wall transforms into an intricate 3D architecture, composed of an outer compact layer enveloping an inner layer of multicellular trabecular ridges. How these tissue layers acquire their characteristic form suited for their function remains an open question. Here, we find that multiscale mechanochemical coupling and an emergent tissue-scale morphological transition steer functional maturation of the developing zebrafish heart. Single-celled trabecular seeds recruit outer compact layer cells to mature into clonally heterogenous multicellular ridges, thereby amplifying cardiac contractile forces. In response, remaining compact layer cells are stretched, which impedes their further recruitment, thereby constraining trabecular ridge density. Concomitantly, Notch-dependent actomyosin dampening triggers a sharp transition in myocardial tissue area, activating rapid organ growth that expands blood filling capacity. Thus, multiscale self-organizing interactions optimize heart size and contractile efficiency to support embryonic life.
]]></description>
<dc:creator>Andrews, T. G.</dc:creator>
<dc:creator>Cornwall Scoones, J.</dc:creator>
<dc:creator>Ramel, M.-C.</dc:creator>
<dc:creator>Gupta, K.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Priya, R.</dc:creator>
<dc:date>2024-07-24</dc:date>
<dc:identifier>doi:10.1101/2024.07.24.604962</dc:identifier>
<dc:title><![CDATA[Multiscale mechanics drive functional maturation of the vertebrate heart]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.24.604964v1?rss=1">
<title>
<![CDATA[
ETV4 and ETV5 Orchestrate FGF-Mediated Lineage Specification and Epiblast Maturation during Early Mouse Development 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.24.604964v1?rss=1"
</link>
<description><![CDATA[
Cell fate decisions in early mammalian embryos are tightly regulated processes crucial for proper development. While FGF signaling plays key roles in early embryo patterning, its downstream effectors remain poorly understood. Our study demonstrates that the transcription factors Etv4 and Etv5 are critical mediators of FGF signaling in cell lineage specification and maturation in mouse embryos. We show that loss of Etv5 compromises primitive endoderm formation at pre-implantation stages. Furthermore, Etv4/5 deficiency delays naive pluripotency exit and epiblast maturation, leading to elevated NANOG and reduced OTX2 expression within the blastocyst epiblast. As a consequence of delayed pluripotency progression, Etv4/5 deficient embryos exhibit anterior visceral endoderm migration defects post-implantation, a process essential for coordinated embryonic patterning and gastrulation initiation. Our results demonstrate the successive roles of these FGF signaling effectors in early lineage specification and embryonic body plan establishment, providing new insights into the molecular control of mammalian development.

Summary statementFGF signaling effectors ETV4/5 regulate lineage specification and embryo patterning in mice, affecting primitive endoderm formation and pluripotency exit.
]]></description>
<dc:creator>Simon, C. S.</dc:creator>
<dc:creator>Garg, V.</dc:creator>
<dc:creator>Kuo, Y.-Y.</dc:creator>
<dc:creator>Niakan, K. K.</dc:creator>
<dc:creator>Hadjantonakis, A.-K.</dc:creator>
<dc:date>2024-07-24</dc:date>
<dc:identifier>doi:10.1101/2024.07.24.604964</dc:identifier>
<dc:title><![CDATA[ETV4 and ETV5 Orchestrate FGF-Mediated Lineage Specification and Epiblast Maturation during Early Mouse Development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.25.605091v1?rss=1">
<title>
<![CDATA[
2'-O-Methyl-guanosine 3-base RNA fragments mediate essential natural TLR7/8 antagonism 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.25.605091v1?rss=1"
</link>
<description><![CDATA[
Recognition of RNA fragments by Toll-like receptors (TLR) 7 and 8 is a key contributor to the initiation of a protective innate immune response against pathogens. A long-standing enigma is how degradation products of host RNAs, generated by the daily phagocytic clearance of billions of apoptotic cells, fail to activate TLR7 and TLR8 signalling1. Here, we report that select 2-O-methyl (2-Ome) guanosine RNA fragments as short as 3 bases, including those derived from host-RNAs, are potent TLR7 and TLR8 antagonists that reduce TLR7 sensing in vivo. Mechanistically, antagonistic fragments are directed towards a distinct binding site on these proteins by 5-end 2-Ome guanosine. Our results indicate that host-RNAs evade detection by TLR7/8 due to a pool of abundant host ribosomal 2-Ome-modified RNA fragments that naturally antagonize TLR7 and TLR8 sensing to avoid auto-immunity. Crucially, rare TLR7 and TLR8 mutations located at this antagonistic site decrease the inhibitory activity of 2-Ome guanosine RNA fragments and lead to auto-immunity in patients. Our findings also establish that select chemically synthesised 3-base oligonucleotides can harness the protective anti-inflammatory activity of this natural immune checkpoint for therapeutic targeting of TLR7-driven diseases.

One Sentence SummaryShort 2-O-Methyl RNA fragments are natural TLR7/8 antagonists
]]></description>
<dc:creator>Alharbi, A.</dc:creator>
<dc:creator>Sapkota, S.</dc:creator>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Jin, R.</dc:creator>
<dc:creator>Jayasekara, W. S.</dc:creator>
<dc:creator>Rupasinghe, E.</dc:creator>
<dc:creator>Speir, M.</dc:creator>
<dc:creator>Wilkinson-White, L.</dc:creator>
<dc:creator>Gamsjaeger, R.</dc:creator>
<dc:creator>Cubeddu, L.</dc:creator>
<dc:creator>Ellyard, J. I.</dc:creator>
<dc:creator>Wenholz, D.</dc:creator>
<dc:creator>McAllan, A.</dc:creator>
<dc:creator>Rezwan, R.</dc:creator>
<dc:creator>Ying, L.</dc:creator>
<dc:creator>Hosseini Far, H.</dc:creator>
<dc:creator>Bones, J.</dc:creator>
<dc:creator>He, S.</dc:creator>
<dc:creator>Yu, D.</dc:creator>
<dc:creator>Lennox, K.</dc:creator>
<dc:creator>Hertzog, P. J.</dc:creator>
<dc:creator>Vinuesa, C.</dc:creator>
<dc:creator>Behlke, M. A.</dc:creator>
<dc:creator>Ohto, U.</dc:creator>
<dc:creator>Laczka, O.</dc:creator>
<dc:creator>Corry, B.</dc:creator>
<dc:creator>Shimizu, T.</dc:creator>
<dc:creator>Gantier, M. P.</dc:creator>
<dc:date>2024-07-25</dc:date>
<dc:identifier>doi:10.1101/2024.07.25.605091</dc:identifier>
<dc:title><![CDATA[2'-O-Methyl-guanosine 3-base RNA fragments mediate essential natural TLR7/8 antagonism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.25.605137v1?rss=1">
<title>
<![CDATA[
Robust proteome profiling of cysteine-reactive fragments using label-free chemoproteomics 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.25.605137v1?rss=1"
</link>
<description><![CDATA[
Identifying pharmacological probes for human proteins represents a key opportunity to accelerate the discovery of new therapeutics. High-content screening approaches to expand the ligandable proteome offer the potential to expedite the discovery of novel chemical probes to study protein function. Screening libraries of reactive fragments by chemoproteomics offers a compelling approach to ligand discovery, however, optimising sample throughput, proteomic depth, and data reproducibility remains a key challenge.

We report a versatile, label-free quantification proteomics platform for competitive profiling of cysteine-reactive fragments against the native proteome. This high-throughput platform combines SP4 plate-based sample preparation with rapid chromatographic gradients. Data-independent acquisition performed on a Bruker timsTOF Pro 2 consistently identified [~]23,000 cysteine sites per run, with a total of [~]32,000 cysteine sites profiled in HEK293T and Jurkat lysate. Crucially, this depth in cysteinome coverage is met with high data completeness, enabling robust identification of liganded proteins.

In this study, 80 reactive fragments were screened in two cell lines identifying >400 ligand-protein interactions. Hits were validated through concentration-response experiments and the platform was utilised for hit expansion and live cell experiments. This label-free platform represents a significant step forward in high-throughput proteomics to evaluate ligandability of cysteines across the human proteome.
]]></description>
<dc:creator>Biggs, G. S.</dc:creator>
<dc:creator>Cawood, E. E.</dc:creator>
<dc:creator>Vuorinen, A.</dc:creator>
<dc:creator>McCarthy, W. J.</dc:creator>
<dc:creator>Wilders, H.</dc:creator>
<dc:creator>Riziotis, I. G.</dc:creator>
<dc:creator>van der Zouwen, A. J.</dc:creator>
<dc:creator>Pettinger, J.</dc:creator>
<dc:creator>Nightingale, L.</dc:creator>
<dc:creator>Chen, P.</dc:creator>
<dc:creator>Powell, A. J.</dc:creator>
<dc:creator>House, D.</dc:creator>
<dc:creator>Boulton, S. J.</dc:creator>
<dc:creator>Skehel, J. M.</dc:creator>
<dc:creator>Rittinger, K.</dc:creator>
<dc:creator>Bush, J. T.</dc:creator>
<dc:date>2024-07-25</dc:date>
<dc:identifier>doi:10.1101/2024.07.25.605137</dc:identifier>
<dc:title><![CDATA[Robust proteome profiling of cysteine-reactive fragments using label-free chemoproteomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.30.605801v1?rss=1">
<title>
<![CDATA[
Human-like APOBEC3 gene expression and anti-viral responses following replacement of mouse Apobec3 with the 7-gene human APOBEC3 locus 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.30.605801v1?rss=1"
</link>
<description><![CDATA[
The seven human APOBEC3 (hA3) genes encode polynucleotide cytidine deaminases that play vital roles in restricting replication of viruses and retrotransposons. However, off-target A3 deamination of the cellular genome is a major source of somatic mutations in human cancer. The ability to study A3 biology in vivo is hindered by the fact that the solitary murine Apobec3 gene (mA3) encodes a cytoplasmic enzyme, with no apparent mutagenic activity. Transgenic expression of individual hA3 genes in mice has helped to confirm their oncogenic potential but important questions including which hA3 genes are active in different tissue contexts and how they function in concert when under control of their cognate promoters cannot be addressed using these models. Here we describe humanization of the mouse mA3 locus by integration of a modified BAC clone encompassing the entire 7-gene hA3 locus from human chromosome 22 replacing mA3 on mouse chromosome 15. APOBEC3 mice are viable and fertile and hA3 gene expression in cells and tissues correlates strongly with expression in corresponding human cells and tissues, indicating human-like regulation of hA3 gene expression in the mice. Splenocytes from this line display a functional human A3 response to Friend Murine Leukaemia Virus (F-MLV) infection. We propose that the Hs-APOBEC3 mouse will uniquely model the function of the complete hA3 locus in a living organism and that it will serve as a useful background upon which to model human cancer, as well as assisting drug discovery efforts.
]]></description>
<dc:creator>Kirkwood, N. K.</dc:creator>
<dc:creator>Prosser, H.</dc:creator>
<dc:creator>Yap, M. W.</dc:creator>
<dc:creator>Gibson, J.</dc:creator>
<dc:creator>Cook, R.</dc:creator>
<dc:creator>Reddin, I.</dc:creator>
<dc:creator>Stranger, A.</dc:creator>
<dc:creator>Howes, E.</dc:creator>
<dc:creator>Zainal, N.</dc:creator>
<dc:creator>Periyasamy, M.</dc:creator>
<dc:creator>Thomas, G. J.</dc:creator>
<dc:creator>Scott, J.</dc:creator>
<dc:creator>Bishop, K. N.</dc:creator>
<dc:creator>Ali, S.</dc:creator>
<dc:creator>Bradley, A.</dc:creator>
<dc:creator>Fenton, T. R.</dc:creator>
<dc:date>2024-07-30</dc:date>
<dc:identifier>doi:10.1101/2024.07.30.605801</dc:identifier>
<dc:title><![CDATA[Human-like APOBEC3 gene expression and anti-viral responses following replacement of mouse Apobec3 with the 7-gene human APOBEC3 locus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.01.604391v1?rss=1">
<title>
<![CDATA[
Protein degradation shapes developmental tempo in mouse and human neural progenitors 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.01.604391v1?rss=1"
</link>
<description><![CDATA[
The pace of embryonic development differs markedly across mammalian species, yet the molecular mechanisms underlying these tempo differences remain largely unknown. Here, we systematically compared protein dynamics in mouse and human neural progenitors (NPs) and examined how protein stability influences developmental timing. We find that mouse NPs exhibit faster protein production and degradation than human NPs. Human NPs display broadly increased protein half-lives, independent of cellular compartment or protein function, and this difference persists in post-mitotic neurons. Consistent with this, proteasomal activity is lower in human embryonic spinal cord and stem cell-derived neural progenitors than mouse, correlating with reduced expression of proteasome-associated proteins. Functionally, accelerating the degradation of the key transcriptional repressor IRX3 in mouse NPs speeds the activation of its target gene, providing causal evidence that protein turnover modulates developmental tempo. These results reveal that species-specific regulation of protein degradation shapes the timing of neural development and suggest that evolutionary tuning of proteasomal activity contributes to differences in embryonic developmental pace.
]]></description>
<dc:creator>Nakanoh, S.</dc:creator>
<dc:creator>Stamataki, D.</dc:creator>
<dc:creator>Garcia-Perez, L.</dc:creator>
<dc:creator>Azzi, C.</dc:creator>
<dc:creator>Carr, H. L.</dc:creator>
<dc:creator>Pokhilko, A.</dc:creator>
<dc:creator>Howell, S.</dc:creator>
<dc:creator>Yu, L.</dc:creator>
<dc:creator>Skehel, M.</dc:creator>
<dc:creator>Oxley, D.</dc:creator>
<dc:creator>Andrews, S.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Rayon, T.</dc:creator>
<dc:date>2024-08-01</dc:date>
<dc:identifier>doi:10.1101/2024.08.01.604391</dc:identifier>
<dc:title><![CDATA[Protein degradation shapes developmental tempo in mouse and human neural progenitors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.08.604602v1?rss=1">
<title>
<![CDATA[
Biallelic Loss of Molecular Chaperone Molecule AIP Results in a Novel Severe Multisystem Disease Defined by Defective Proteostasis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.08.604602v1?rss=1"
</link>
<description><![CDATA[
Children born with deleterious biallelic variants of the chaperone aryl hydrocarbon receptor interacting protein (AIP) have a novel pediatric metabolic disease presenting a severe, complex clinical phenotype characterized by failure to develop following birth. Analysis of Aip knockout mouse embryonic fibroblasts and patient-derived dermal fibroblasts revealed that AIP was required to support proteostasis; including proteasome activity, induction of autophagy and lysosome function. aip knockout zebrafish, recapitulated the phenotype of the children; dying at an early stage of development when autophagy is required to adapt to periods of starvation. Our results demonstrate that AIP plays a crucial role in initiating autophagy and maintaining proteostasis in vitro and in vivo.

One Sentence SummaryHomozygous loss of the chaperone AIP results in a novel pediatric disease exhibiting multiple features of a lysosomal storage disease.
]]></description>
<dc:creator>Korbonits, M.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Barry, S.</dc:creator>
<dc:creator>Lim, C.</dc:creator>
<dc:creator>Suleyman, O.</dc:creator>
<dc:creator>De-Tito, S.</dc:creator>
<dc:creator>Begum, N.</dc:creator>
<dc:creator>Vignola, M. L.</dc:creator>
<dc:creator>Hall, C.</dc:creator>
<dc:creator>Perna, L.</dc:creator>
<dc:creator>Chapple, P.</dc:creator>
<dc:creator>Henson, S.</dc:creator>
<dc:creator>Morales, V.</dc:creator>
<dc:creator>Bianchi, K.</dc:creator>
<dc:creator>Edvardsson, V. O.</dc:creator>
<dc:creator>Ragnarsson, K. A.</dc:creator>
<dc:creator>Kristinsdottir, V. E.</dc:creator>
<dc:creator>Debeer, A.</dc:creator>
<dc:creator>Sleyp, Y.</dc:creator>
<dc:creator>Zinchenko, R.</dc:creator>
<dc:creator>Anderson, G.</dc:creator>
<dc:creator>Duchen, M.</dc:creator>
<dc:creator>Singh, K.</dc:creator>
<dc:creator>Chung, C.-Y.</dc:creator>
<dc:creator>Yuan, Y.</dc:creator>
<dc:creator>Patel, S.</dc:creator>
<dc:creator>Aksoy, E.</dc:creator>
<dc:creator>Borovikov, A. O.</dc:creator>
<dc:creator>Bjornsson, H. T.</dc:creator>
<dc:creator>Van Esch, H.</dc:creator>
<dc:creator>Czibik, G.</dc:creator>
<dc:creator>Tooze, S.</dc:creator>
<dc:creator>Brennan, C. H.</dc:creator>
<dc:creator>Haworth, O.</dc:creator>
<dc:date>2024-08-08</dc:date>
<dc:identifier>doi:10.1101/2024.08.08.604602</dc:identifier>
<dc:title><![CDATA[Biallelic Loss of Molecular Chaperone Molecule AIP Results in a Novel Severe Multisystem Disease Defined by Defective Proteostasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.10.607181v1?rss=1">
<title>
<![CDATA[
The eduWOSM: an advanced optical microscope for education 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.10.607181v1?rss=1"
</link>
<description><![CDATA[
To improve access to advanced optical microscopy in educational and resource-limited settings we have developed the eduWOSM (educational Warwick Open Source Microscope), an open hardware platform for transmitted-light and epifluorescence imaging in up to 4 colours, including single molecule imaging. EduWOSMs are robust, bright, compact, portable and ultra-stable. They are controlled entirely by open source hardware and software, with an option for remote control from a webpage. Here we describe the core eduWOSM technology and benchmark its performance using 3 example projects, single fluorophore tracking of tubulin heterodimers within gliding microtubules, 4D (deconvolution) imaging/tracking of chromosome motions in dividing human cells, and automated single particle tracking in vitro and in live cells with classification into subdiffusive, diffusive and superdiffusive motion.
]]></description>
<dc:creator>Carter, N. J.</dc:creator>
<dc:creator>Martin, D. S.</dc:creator>
<dc:creator>Molloy, J. E.</dc:creator>
<dc:creator>Cross, R. A.</dc:creator>
<dc:date>2024-08-10</dc:date>
<dc:identifier>doi:10.1101/2024.08.10.607181</dc:identifier>
<dc:title><![CDATA[The eduWOSM: an advanced optical microscope for education]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.12.606135v1?rss=1">
<title>
<![CDATA[
Results of the Protein Engineering Tournament: An Open Science Benchmark for Protein Modeling and Design 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.12.606135v1?rss=1"
</link>
<description><![CDATA[
The grand challenge of protein engineering is the development of computational models to characterize and generate protein sequences for arbitrary functions. Progress is limited by lack of 1) benchmarking opportunities, 2) large protein function datasets, and 3) access to experimental protein characterization. We introduce the Protein Engineering Tournament--a fully-remote competition designed to foster the development and evaluation of computational approaches in protein engineering. The tournament consists of an in silico round, predicting biophysical properties from protein sequences, followed by an in vitro round where novel protein sequences are designed, expressed and characterized using automated methods. Upon completion, all datasets, experimental protocols, and methods are made publicly available. We detail the structure and outcomes of a pilot Tournament involving seven protein design teams, powered by six multi-objective datasets, with experimental characterization by our partner, International Flavors and Fragrances. Forthcoming Protein Engineering Tournaments aim to mobilize the scientific community towards transparent evaluation of progress in the field.

Graphical Abstract

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]]></description>
<dc:creator>Armer, C.</dc:creator>
<dc:creator>Kane, H.</dc:creator>
<dc:creator>Cortade, D. L.</dc:creator>
<dc:creator>Redestig, H.</dc:creator>
<dc:creator>Estell, D. A.</dc:creator>
<dc:creator>Yusuf, A.</dc:creator>
<dc:creator>Rollins, N. J.</dc:creator>
<dc:creator>Spinner, H.</dc:creator>
<dc:creator>Marks, D.</dc:creator>
<dc:creator>Brunette, T.</dc:creator>
<dc:creator>Kelly, P. J.</dc:creator>
<dc:creator>DeBenedictis, E.</dc:creator>
<dc:date>2024-08-12</dc:date>
<dc:identifier>doi:10.1101/2024.08.12.606135</dc:identifier>
<dc:title><![CDATA[Results of the Protein Engineering Tournament: An Open Science Benchmark for Protein Modeling and Design]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.13.607563v1?rss=1">
<title>
<![CDATA[
Arp2/3 iso-complexes limit t-tubule overgrowth to ensure myofiber formation and synchronous contraction 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.13.607563v1?rss=1"
</link>
<description><![CDATA[
Muscle cells form a network of plasma membrane invaginations called t-tubules that control calcium release from the endoplasmic reticulum at the triads during muscle contraction. Although the importance of t-tubules for muscle physiology is well established, and abnormalities are found in multiple disorders, the mechanisms that mediate t-tubule growth are unknown. We show that the actomyosin cortex beneath the plasma membrane, regulated by Arp2/3 complexes containing Arpc5, acts as a gatekeeper for the membrane availability required for t-tubule growth. Enlarged t-tubules are formed upon disruption of Arpc5, impairing the synchronisation between plasma membrane depolarization and calcium release. Additionally, ablation of Arpc5 postnatally in myofibers results in muscle fatigue and t-tubule abnormalities, as observed in muscle disorders. We propose that the actomyosin cortex impacts muscle function, offering a potential pathophysiological mechanism for muscle disorders.
]]></description>
<dc:creator>Pereira, A. R.</dc:creator>
<dc:creator>Soares, A. d. R.</dc:creator>
<dc:creator>Liu, T.</dc:creator>
<dc:creator>Ferreira, J. L.</dc:creator>
<dc:creator>Leal, G.</dc:creator>
<dc:creator>Faleiro, I.</dc:creator>
<dc:creator>Kogata, N.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:creator>Moores, C.</dc:creator>
<dc:creator>Gomes, E. R.</dc:creator>
<dc:date>2024-08-13</dc:date>
<dc:identifier>doi:10.1101/2024.08.13.607563</dc:identifier>
<dc:title><![CDATA[Arp2/3 iso-complexes limit t-tubule overgrowth to ensure myofiber formation and synchronous contraction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.13.607787v1?rss=1">
<title>
<![CDATA[
A Biologically Inspired Attention Model for Neural Signal Analysis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.13.607787v1?rss=1"
</link>
<description><![CDATA[
Understanding how the brain represents sensory information and triggers behavioural responses is a fundamental goal in neuroscience. Recent advances in neuronal recording techniques aim to progress towards this milestone, yet the resulting high-dimensional responses are challenging to interpret and link to relevant variables. Although existing machine learning models propose to do so, they often sacrifice interpretability for predictive power, effectively operating as black boxes. In this work, we introduce SPARKS, a biologically inspired model capable of high decoding accuracy and interpretable discovery within a single framework. SPARKS adapts the self-attention mechanism of large language models to extract information from the timing of single spikes and the sequence in which neurons fire using Hebbian learning. Trained with a criterion inspired by predictive coding to enforce temporal coherence, our model produces low-dimensional latent embeddings that are robust across sessions and animals. By directly capturing the underlying data distribution through a generative encoding-decoding framework, SPARKS exhibits state-of-the-art predictive capabilities across diverse electrophysiology and calcium imaging datasets from the motor, visual and entorhinal cortices. Crucially, the Hebbian coefficients learned by the model are interpretable, allowing us to infer the effective connectivity and recover the known functional hierarchy of the mouse visual cortex. Overall, SPARKS unifies representation learning, high-performance decoding and model interpretability in a single framework by bridging neuroscience and AI, providing a powerful and versatile tool for dissecting neural computations and marking a step towards the next generation of biologically inspired intelligent systems.
]]></description>
<dc:creator>Skatchkovsky, N.</dc:creator>
<dc:creator>Glazman, N.</dc:creator>
<dc:creator>Sadeh, S.</dc:creator>
<dc:creator>Iacaruso, F.</dc:creator>
<dc:date>2024-08-16</dc:date>
<dc:identifier>doi:10.1101/2024.08.13.607787</dc:identifier>
<dc:title><![CDATA[A Biologically Inspired Attention Model for Neural Signal Analysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.13.607853v1?rss=1">
<title>
<![CDATA[
Early atrial remodelling drives arrhythmia in Fabry Disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.13.607853v1?rss=1"
</link>
<description><![CDATA[
BackgroundFabry disease (FD) is an X-linked lysosomal storage disorder caused by -galactosidase A (-Gal A) deficiency, resulting in multi-organ accumulation of sphingolipid, namely globotriaosylceramide (Gb3). This triggers ventricular myocardial hypertrophy, fibrosis, and inflammation, driving arrhythmia and sudden death, a common cause of FD mortality. Atrial fibrillation (AF) is common in FD, yet the cellular mechanisms accounting for this are unknown. To address this, we conducted electrocardiography (ECG) analysis from a large cohort of adults with FD at varying stages of cardiomyopathy. Cellular contractile and electrophysiological function were examined in an atrial FD model, developed using gene-edited atrial cardiomyocytes and imputed into in-silico atrial models to provide insight into arrhythmia mechanisms.

MethodsIn 115 adults with FD, ECG P-wave characteristics were compared with non-FD controls. Induced pluripotent stem cells (iPSCs) were genome-edited using CRISPR-Cas9 to introduce the GLA p. N215S variant and differentiated into atrial cardiomyocytes (iPSC-CMs). Contraction, calcium handling and electrophysiology experiments were conducted to explore proarrhythmic mechanisms. A bi-atrial in-silico model was developed with the cellular changes induced by GLA p. N215S iPSC-CMs.

ResultsECG analysis demonstrated P-wave duration and PQ interval shortening in FD adults before onset of cardiomyopathy on imaging and biochemical criteria. FD patients exhibited a higher incidence of premature atrial contractions and increased risk of developing AF. In our cellular model, GLA p. N215S iPSC-CMs were deficient in -Gal A and exhibited Gb3 accumulation. Atrial GLA p. N215S iPSC-CMs demonstrated a more positive diastolic membrane potential, faster action potential upstroke velocity, greater burden of delayed afterdepolarizations, greater contraction force, slower beat rate and dysfunction in calcium handling compared to wildtype iPSC-CMs. Inputting these changes into the in-silico model resulted in similar P-wave morphology changes to those seen in early FD cardiomyopathy and increased the action potential duration (APD) restitution slope, causing APD alternans and inducing AF.

ConclusionsThese findings enhance our understanding of atrial myopathy in FD by providing novel insights into underpinning mechanisms for atrial arrhythmia and a rationale for early P-wave changes. These may be targeted in future research to develop therapeutic strategies to reduce the arrhythmic burden in FD and other atrial cardiomyopathies.
]]></description>
<dc:creator>Roy, A.</dc:creator>
<dc:creator>O'Shea, C.</dc:creator>
<dc:creator>Dasi, A.</dc:creator>
<dc:creator>Patel, L.</dc:creator>
<dc:creator>CUMBERLAND, M.</dc:creator>
<dc:creator>NIEVES, D.</dc:creator>
<dc:creator>Canagarajah, H. S.</dc:creator>
<dc:creator>THOMPSON, S.</dc:creator>
<dc:creator>AZAD, A.</dc:creator>
<dc:creator>Price, A. M.</dc:creator>
<dc:creator>HALL, C.</dc:creator>
<dc:creator>Alvior, A. M. B.</dc:creator>
<dc:creator>RATH, P.</dc:creator>
<dc:creator>DAVIES, B.</dc:creator>
<dc:creator>Rodriguez, B.</dc:creator>
<dc:creator>Holmes, A. P.</dc:creator>
<dc:creator>Pavlovic, D.</dc:creator>
<dc:creator>Townend, J. N.</dc:creator>
<dc:creator>Geberhiwot, T.</dc:creator>
<dc:creator>GEHMLICH, K.</dc:creator>
<dc:creator>Steeds, R. P.</dc:creator>
<dc:date>2024-08-17</dc:date>
<dc:identifier>doi:10.1101/2024.08.13.607853</dc:identifier>
<dc:title><![CDATA[Early atrial remodelling drives arrhythmia in Fabry Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.21.608818v1?rss=1">
<title>
<![CDATA[
Horizontal gene transfer-initiated reorganization of lipid metabolism drives lifestyle innovation in a eukaryote 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.21.608818v1?rss=1"
</link>
<description><![CDATA[
Horizontal gene transfer is a source of metabolic innovation and adaptation to new environments. Yet, how horizontally transferred metabolic functionalities are integrated into host cell biology remains an open question. Here, we use the fission yeast Schizosaccharomyces japonicus to probe how eukaryotic lipid metabolism is rewired in response to the acquisition of a horizontally transferred squalene-hopene cyclase Shc1. We show that Shc1-dependent production of hopanoids, the structural mimics of eukaryotic sterols, allows S. japonicus to thrive in anoxia, where sterol biosynthesis is not possible. We further demonstrate that glycerophospholipid fatty acyl asymmetry, prevalent in S. japonicus, is crucial for accommodating both sterols and hopanoids in membranes, and explain how Shc1 functions alongside the native sterol biosynthetic pathway to support membrane properties. Through engineering experiments in the sister species S. pombe, which naturally lacks Shc1, we show that the acquisition of Shc1 may entail new physiological traits; however, to maximize Shc1 performance, sterol biosynthesis must be dampened. Our work sheds new light on the mechanisms underlying cellular integration of horizontally transferred genes in eukaryotes and provides broader insights into the evolution of membrane organization and function.
]]></description>
<dc:creator>Rao, B. D.</dc:creator>
<dc:creator>Gomez Gil, E.</dc:creator>
<dc:creator>Peter, M.</dc:creator>
<dc:creator>Balogh, G.</dc:creator>
<dc:creator>Nunes, V.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Chen, Q.</dc:creator>
<dc:creator>Rosenthal, P. B.</dc:creator>
<dc:creator>Oliferenko, S.</dc:creator>
<dc:date>2024-08-21</dc:date>
<dc:identifier>doi:10.1101/2024.08.21.608818</dc:identifier>
<dc:title><![CDATA[Horizontal gene transfer-initiated reorganization of lipid metabolism drives lifestyle innovation in a eukaryote]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.23.609334v1?rss=1">
<title>
<![CDATA[
Paracrine rescue of MYR1-deficient Toxoplasma gondii mutants reveals limitations of pooled in vivo CRISPR screens 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.23.609334v1?rss=1"
</link>
<description><![CDATA[
Toxoplasma gondii is an intracellular parasite that subverts host cell functions via secreted virulence factors. Up to 70% of parasite-controlled changes in the host transcriptome rely on the MYR1 protein, which is required for the translocation of secreted proteins into the host cell. Mice infected with MYR1 knock-out (KO) strains survive infection, supporting a paramount function of MYR1-dependent secreted proteins in Toxoplasma virulence and proliferation. However, we have previously shown that MYR1 mutants have no growth defect in pooled in vivo CRISPR-Cas9 screens in mice, suggesting that the presence of parasites that are wild-type at the myr1 locus in pooled screens can rescue the phenotype. Here, we demonstrate that MYR1 is not required for the survival in IFN-{gamma}-activated murine macrophages, and that parasites lacking MYR1 are able to expand during the onset of infection. While {Delta}MYR1 parasites have restricted growth in single-strain murine infections, we show that the phenotype is rescued by co-infection with wild-type (WT) parasites in vivo, independent of host functional adaptive immunity or key pro-inflammatory cytokines. These data show that the major function of MYR1-dependent secreted proteins is not to protect the parasite from clearance within infected cells. Instead, MYR-dependent proteins generate a permissive niche in a paracrine manner, which rescues {Delta}MYR1 parasites within a pool of CRISPR mutants in mice. Our results highlight an important limitation of otherwise powerful in vivo CRISPR screens and point towards key functions for MYR1-dependent Toxoplasma-host interactions beyond the infected cell.

SIGNIFICANCE STATEMENTPooled CRISPR screens are powerful tools to interrogate gene function in a high-throughput manner. Genes conferring fitness advantages or disadvantages upon disruption can be identified by sequencing. However, in Toxoplasma gondii pooled CRISPR screens in mice, fitness defects for some selected mutants drastically diverge from those observed in single-strain infections. Here, we show that a growth defect of a single Toxoplasma gene deletion mutant is rescued if co-infected with wildtype parasites. These results shine light on Toxoplasmas ability to subvert the host response beyond the infected cell, and highlight an important limitation of pooled CRISPR screens in mice. This limitation is probably encountered in CRISPR screens in general where paracrine effects occur.
]]></description>
<dc:creator>Torelli, F.</dc:creator>
<dc:creator>Matos da Fonseca, D.</dc:creator>
<dc:creator>Butterworth, S.</dc:creator>
<dc:creator>Young, J. C.</dc:creator>
<dc:creator>Treeck, M.</dc:creator>
<dc:date>2024-08-23</dc:date>
<dc:identifier>doi:10.1101/2024.08.23.609334</dc:identifier>
<dc:title><![CDATA[Paracrine rescue of MYR1-deficient Toxoplasma gondii mutants reveals limitations of pooled in vivo CRISPR screens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.27.596794v1?rss=1">
<title>
<![CDATA[
Synchronous L1 retrotransposition events promote chromosomal crossover early in human tumorigenesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.27.596794v1?rss=1"
</link>
<description><![CDATA[
L1 retrotransposition is a significant source of genomic variation in human epithelial tumours, which can contribute to tumorigenesis. However, fundamental questions about the causes and consequences of L1 activity in cancer genomes remain unresolved, primarily due to the limitations of short-read sequencing technologies. Here, we employ multiplatform sequencing, with an emphasis on long reads, to analyse a fine selection of 10 tumours exhibiting high rates of somatic retrotransposition, encompassing over 6000 events. The analysis of L1 locus-specific single-nucleotide variants reveals a novel panorama of L1 loci activity. Furthermore, examination of the internal structure of somatic L1s uncovers the mechanisms behind their inactivation. A hidden landscape of chromosomal aberrations emerges in the light of long reads, where reciprocal translocations mediated by L1 insertion represent frequent events. Resolution of L1 bridges configuration elucidates the mechanisms of their formation, where typically two independent, but synchronous, somatic L1 insertions drive the reciprocal exchange between non-homologous chromosomes. Timing analyses indicate that L1 retrotransposition is an early driver of chromosomal instability, active before the first whole-genome doubling event. Overall, these findings highlight L1 activity as a more significant contributor to tumour genome plasticity than previously recognized, extending its impact beyond simple insertional mutagenesis.
]]></description>
<dc:creator>Zumalave, S.</dc:creator>
<dc:creator>Santamarina, M.</dc:creator>
<dc:creator>Espasandin, N. P.</dc:creator>
<dc:creator>Garcia-Souto, D.</dc:creator>
<dc:creator>Temes, J.</dc:creator>
<dc:creator>Baker, T. M.</dc:creator>
<dc:creator>Valtierra, A. P.</dc:creator>
<dc:creator>Otero, I.</dc:creator>
<dc:creator>Rodriguez-Castro, J.</dc:creator>
<dc:creator>Oitaben, A.</dc:creator>
<dc:creator>Alvarez, E. G.</dc:creator>
<dc:creator>Otero, P.</dc:creator>
<dc:creator>Diaz-Arias, I.</dc:creator>
<dc:creator>Martinez-Fernandez, M.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>Cristofari, G.</dc:creator>
<dc:creator>Rodriguez-Martin, B.</dc:creator>
<dc:creator>Tubio, J. M.</dc:creator>
<dc:date>2024-08-27</dc:date>
<dc:identifier>doi:10.1101/2024.08.27.596794</dc:identifier>
<dc:title><![CDATA[Synchronous L1 retrotransposition events promote chromosomal crossover early in human tumorigenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.28.610038v1?rss=1">
<title>
<![CDATA[
CDKL5's role in microtubule-based transport and cognitive function 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.28.610038v1?rss=1"
</link>
<description><![CDATA[
Cyclin-dependent kinase like 5 (CDKL5) is a serine-threonine kinase highly enriched in mammalian neurons. CDKL5 is located on the X-chromosome and its loss-of-function leads to a severe neurodevelopmental disorder called CDKL5 deficiency disorder (CDD). CDKL5 phosphorylates microtubule-associated protein MAP1S and regulates its binding to microtubules. How MAP1S phosphorylation affects microtubule function is not well understood. To address this question, we generated MAP1S phosphomutant mice, in which the CDKL5 phosphorylation sites S786 and S812 are mutated to Alanine (MAP1S S786/812A or MAP1S SA). Using a microtubule co-sedimentation assay, we showed that dynein binding to microtubules is severely reduced in CDKL5 knockout (KO) and MAP1S SA brains. Time-lapse imaging in primary neurons showed impaired dynein motility in both Cdkl5 KO and MAP1S SA. Dynein-driven cargo transport was affected in mutant neuron dendrites, including the delivery of -amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors. We next studied tubulin tyrosination in Cdkl5 KO and MAP1S SA neurons and found that both mutants had a reduced tubulin tyrosination when compared to WT neurons. Since dynein-dynactin has a higher affinity for tyrosinated microtubules, we hypothesized that reduced tyrosination in MAP1S phosphomutant mice could be the mechanistic cause of impaired dynein motility. In support of this, we show that upon expression of tubulin tyrosine kinase TTL, we rescued dynein motility defects in MAP1S phosphomutant neurons. Hippocampal neurons derived from MAP1S SA mice revealed a significant reduction in spine density and synapses, and altered spine morphology. Finally, behavioral phenotyping of MAP1S phosphomutant mice showed increased anxiety, impaired motor performance, social and memory deficits, mirroring to some extent the clinical manifestations present in CDD patients. Our results reveal MAP1S phosphorylation to be an important contributor to dynein-mediated transport and synapse formation.

Graphical abstract

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]]></description>
<dc:creator>Lopes, A. T.</dc:creator>
<dc:creator>Janiv, O.</dc:creator>
<dc:creator>Claxton, S.</dc:creator>
<dc:creator>Ultanir, S. K.</dc:creator>
<dc:date>2024-08-29</dc:date>
<dc:identifier>doi:10.1101/2024.08.28.610038</dc:identifier>
<dc:title><![CDATA[CDKL5's role in microtubule-based transport and cognitive function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.30.610462v1?rss=1">
<title>
<![CDATA[
M. tuberculosis invades and disrupts the blood-brain barrier directly to initiate meningitis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.30.610462v1?rss=1"
</link>
<description><![CDATA[
Tuberculous meningitis (TBM) is the most severe form of tuberculosis and HIV-1 co-infection worsens the already poor prognosis. How Mycobacterium tuberculosis (Mtb) crosses the blood brain barrier (BBB) and the influence of HIV-1 on pathogenesis remains unclear. Using human pericytes, astrocytes, endothelial cells, and microglia alone; and combined in an in vitro BBB we investigated Mtb +/- HIV-1 co-infection on central nervous system cell entry and function. Mtb infected and multiplied in all cell types with HIV-1 increasing entry to astrocytes and pericytes, and growth in HIV-1 positive pericytes and endothelial cells. The permeability of the BBB increased resulting in translocation of bacilli across it. Cytopathic effects included increased markers of cellular stress, ROS release, the induction of neurotoxic astrocytes, and increased secretion of neuroexcitotoxic glutamate. Distinct cell-type specific production of inflammatory and effector mediators were observed. These data indicate Mtb can translocate the BBB directly to initiate meningitis.
]]></description>
<dc:creator>Proust, A.</dc:creator>
<dc:creator>Wilkinson, K. A.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:date>2024-08-30</dc:date>
<dc:identifier>doi:10.1101/2024.08.30.610462</dc:identifier>
<dc:title><![CDATA[M. tuberculosis invades and disrupts the blood-brain barrier directly to initiate meningitis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.03.610831v1?rss=1">
<title>
<![CDATA[
Malaria parasites undergo a rapid and extensive metamorphosis after invasion of the host erythrocyte 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.03.610831v1?rss=1"
</link>
<description><![CDATA[
Within the human host, the symptoms of malaria are caused by the replication of malaria parasites within erythrocytes. Growth inside the erythrocyte exposes the parasites to the normal surveillance of erythrocytes by the host organism, in particular the clearance of erythrocytes in the spleen. Here we show that the malaria parasite Plasmodium falciparum undergoes a rapid, multi-step metamorphosis that transforms the invasive merozoite into an amoeboid-shaped cell within minutes after invading erythrocytes. This transformation involves an increase in the parasite surface area and is mediated by factors already present in the merozoite, including the parasite phospholipid transfer protein PV6. Parasites lacking PV6 do not assume an amoeboid form and instead are spherical and have a smaller surface area than amoeboid forms. Furthermore, erythrocytes infected with parasites lacking PV6 undergo a higher loss of surface area upon infection by P. falciparum, which affects the traversal of infected erythrocytes through the spleen. This is the first evidence that after invasion, the parasite undergoes a rapid, complex metamorphosis within the host erythrocyte that promotes survival in the host.
]]></description>
<dc:creator>Freville, A.</dc:creator>
<dc:creator>Moreira-Leite, F.</dc:creator>
<dc:creator>Roussel, C.</dc:creator>
<dc:creator>Russell, M. R. G.</dc:creator>
<dc:creator>Fricot, A.</dc:creator>
<dc:creator>Carret, V.</dc:creator>
<dc:creator>Sissoko, A.</dc:creator>
<dc:creator>Hayes, M. J.</dc:creator>
<dc:creator>Dialo, A. B.</dc:creator>
<dc:creator>Kerkhoven, N.</dc:creator>
<dc:creator>Ressurreicao, M.</dc:creator>
<dc:creator>Dokmak, S.</dc:creator>
<dc:creator>Blackman, M.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:creator>Buffet, P. A.</dc:creator>
<dc:creator>Vaughan, S.</dc:creator>
<dc:creator>Ndour, A.</dc:creator>
<dc:creator>van Ooij, C.</dc:creator>
<dc:date>2024-09-03</dc:date>
<dc:identifier>doi:10.1101/2024.09.03.610831</dc:identifier>
<dc:title><![CDATA[Malaria parasites undergo a rapid and extensive metamorphosis after invasion of the host erythrocyte]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.04.610994v1?rss=1">
<title>
<![CDATA[
Subtype-specific Patterns of Evolution and Clinically Relevant Genomic Instability in Wilms Tumour 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.04.610994v1?rss=1"
</link>
<description><![CDATA[
BackgroundUnderstanding cancer evolution is fundamental to predicting cancer progression. However, the evolution of paediatric cancers is still under-researched. Large cohorts of patients are required to determine consistent evolutionary trajectories that shed light on key steps in cancer development and reveal underlying biology, especially in rare cancers. Additionally, well annotated clinical data is necessary for determining if evolutionary biomarkers are predictive of patient outcome.

MethodsWe performed detailed evolutionary analysis of 64 paediatric kidney cancers, including 60 Wilms tumours (WT), using DNA microarrays and, in a subset of 30 patients, a WT-specific targeted sequencing assay, to detect copy number alterations (CNA) and mutations, respectively. By analysing multiple tissue samples in the majority of cases we could detect mutation heterogeneity in each tumour. We reconstructed clones across the cohort and described their phylogenetic histories, in addition to detecting mirrored subclonal allelic imbalance.

ResultsOur results highlight pervasive evidence of parallel evolution in WTs affecting CNAs, and CTNNB1 and TP53 mutations. Furthermore, we demonstrate that stromal-type WTs often evolve from a consistent series of events (WT1 mutation, 11p uniparental disomy and CTNNB1 mutation) and we suggest that 19q uniparental disomy is an important ancestral event in both epithelial and diffuse anaplastic WTs. Finally, we propose the total number of evolutionary CNA events as a prognostic biomarker in WTs for event-free survival, particularly in high-risk WT.

ConclusionsOverall, this study sheds light on the evolution of the most common paediatric kidney cancer and links evolutionary analysis to fundamental clinical and biological questions in a large cohort of WTs. We conclude that histological subtypes of WT are often defined by consistent evolutionary sequences. We present evidence that a key marker of evolvability, namely CNA diversity measured phylogenetically across multiple tumour sites, is prognostic of patient outcome and should be considered for clinical use to detect the most aggressive blastemal and diffuse anaplastic type WTs.
]]></description>
<dc:creator>Cresswell, G. D.</dc:creator>
<dc:creator>Chagtai, T.</dc:creator>
<dc:creator>Al-Saadi, R.</dc:creator>
<dc:creator>Treger, T. D.</dc:creator>
<dc:creator>Madhan, G.</dc:creator>
<dc:creator>Mifsud, B.</dc:creator>
<dc:creator>Vujanic, G.</dc:creator>
<dc:creator>Williams, R. D.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:creator>Pritchard-Jones, K.</dc:creator>
<dc:creator>Mifsud, W.</dc:creator>
<dc:date>2024-09-07</dc:date>
<dc:identifier>doi:10.1101/2024.09.04.610994</dc:identifier>
<dc:title><![CDATA[Subtype-specific Patterns of Evolution and Clinically Relevant Genomic Instability in Wilms Tumour]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.06.611651v1?rss=1">
<title>
<![CDATA[
Human DNA replication initiation sites are specified epigenetically by oxidation of 5-methyl-deoxycytidine 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.06.611651v1?rss=1"
</link>
<description><![CDATA[
DNA replication initiates at tens of thousands of sites on the human genome during each S phase. However, no consensus DNA sequence has been found that specifies the locations of these replication origins. Here, we investigate modifications of human genomic DNA by density equilibrium centrifugation and DNA sequencing. We identified short discrete sites with increased density during quiescence and G1 phase that overlap with DNA replication origins before their activation in S phase. The increased density is due to the oxidation of 5-methyl-deoxycytidines by TET enzymes at GC-rich domains. Reversible inhibition of de novo methylation and of subsequent oxidation of deoxycytidines results in a reversible inhibition of DNA replication and of cell proliferation. Our findings suggest a mechanism for the epigenetic specification and semiconservative inheritance of DNA replication origin sites in human cells that also provides a stable integral DNA replication licence to support once-per-cell cycle control of origin activation.
]]></description>
<dc:creator>Krude, T.</dc:creator>
<dc:creator>Bi, J.</dc:creator>
<dc:creator>Doran, R.</dc:creator>
<dc:creator>Jones, R. A.</dc:creator>
<dc:creator>Smith, J. C.</dc:creator>
<dc:date>2024-09-07</dc:date>
<dc:identifier>doi:10.1101/2024.09.06.611651</dc:identifier>
<dc:title><![CDATA[Human DNA replication initiation sites are specified epigenetically by oxidation of 5-methyl-deoxycytidine]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.09.610834v1?rss=1">
<title>
<![CDATA[
Gonadotrophs have a dual origin, with most derived from pituitary stem cells during minipuberty. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.09.610834v1?rss=1"
</link>
<description><![CDATA[
Gonadotrophs are the essential pituitary endocrine cells for reproduction. They produce both luteinizing (LH) and follicle-stimulating (FSH) hormones that act on the gonads. Gonadotrophs first appear in the embryonic pituitary, along with other endocrine cell types, and all expand after birth. We show here that most gonadotrophs originate from a population of postnatal pituitary stem cells during minipuberty, while those generated in the embryo are maintained, revealing an unsuspected dual origin of the adult population. This has implications for our understanding of the establishment and regulation of reproductive functions, both in health and in disease.
]]></description>
<dc:creator>Sheridan, D.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Golan, G.</dc:creator>
<dc:creator>Shiakola, Y.</dc:creator>
<dc:creator>Galichet, C.</dc:creator>
<dc:creator>Mollard, P.</dc:creator>
<dc:creator>Melamed, P.</dc:creator>
<dc:creator>Lovell-Badge, R.</dc:creator>
<dc:creator>Rizzoti, K.</dc:creator>
<dc:date>2024-09-09</dc:date>
<dc:identifier>doi:10.1101/2024.09.09.610834</dc:identifier>
<dc:title><![CDATA[Gonadotrophs have a dual origin, with most derived from pituitary stem cells during minipuberty.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.10.611481v1?rss=1">
<title>
<![CDATA[
In vivo CRISPR screens identify GRA12 as a transcendent secreted virulence factor across Toxoplasma gondii strains and mouse subspecies 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.10.611481v1?rss=1"
</link>
<description><![CDATA[
Toxoplasma gondii parasites exhibit extraordinary host promiscuity owing to over 250 putative secreted proteins that disrupt host cell functions, enabling parasite persistence. However, most of the known effector proteins are specific to Toxoplasma genotypes or hosts. To identify virulence factors that function across different parasite isolates and mouse strains that differ in susceptibility to infection, we performed systematic pooled in vivo CRISPR-Cas9 screens targeting the Toxoplasma secretome. We identified several proteins required for infection across parasite strains and mouse species, of which the dense granule protein 12 (GRA12) emerged as the most important effector protein during acute infection. GRA12 deletion in IFN{gamma}-activated macrophages results in collapsed parasitophorous vacuoles and increased host cell necrosis, which is partially rescued by inhibiting early parasite egress. GRA12 orthologues from related coccidian parasites, including Neospora caninum and Hammondia hammondi, complement Tg{Delta}GRA12 in vitro, suggesting a common mechanism of protection from immune clearance by their hosts.
]]></description>
<dc:creator>Torelli, F.</dc:creator>
<dc:creator>Butterworth, S. W.</dc:creator>
<dc:creator>Lockyer, E. J.</dc:creator>
<dc:creator>Song, O.-R.</dc:creator>
<dc:creator>Pearson-Farr, J.</dc:creator>
<dc:creator>Treeck, M.</dc:creator>
<dc:date>2024-09-10</dc:date>
<dc:identifier>doi:10.1101/2024.09.10.611481</dc:identifier>
<dc:title><![CDATA[In vivo CRISPR screens identify GRA12 as a transcendent secreted virulence factor across Toxoplasma gondii strains and mouse subspecies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.10.611650v1?rss=1">
<title>
<![CDATA[
An integrated stress response-independent role of GCN2 prevents excessive ribosome biogenesis and mRNA translation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.10.611650v1?rss=1"
</link>
<description><![CDATA[
The Integrated Stress Response (ISR) is a corrective physiological program to restore cellular homeostasis that is based on the attenuation of global protein synthesis and a resource-enhancing transcriptional programme. GCN2 is the oldest of four kinases that are activated by diverse cellular stresses to trigger the ISR and acts as the primary responder to amino acid shortage and ribosome collisions. Here, using a broad multi-omic approach, we uncover an ISR-independent role of GCN2. GCN2 inhibition or depletion in the absence of discernible stress causes excessive protein synthesis and ribosome biogenesis, perturbs the cellular translatome, and results in a dynamic and broad loss of metabolic homeostasis. Cancer cells that rely on GCN2 to keep protein synthesis in check under conditions of full nutrient availability depend on GCN2 for survival and unrestricted tumour growth. Our observations define an ISR-independent role of GCN2 in regulating the cellular proteome and translatome and suggest new avenues for cancer therapies based on unleashing excessive mRNA translation.
]]></description>
<dc:creator>Roman-Trufero, M.</dc:creator>
<dc:creator>Kleijn, I. T.</dc:creator>
<dc:creator>Blighe, K.</dc:creator>
<dc:creator>Zhou, J.</dc:creator>
<dc:creator>Saavedra-Garcia, P.</dc:creator>
<dc:creator>Gaffar, A.</dc:creator>
<dc:creator>Christoforou, M.</dc:creator>
<dc:creator>Bellotti, A.</dc:creator>
<dc:creator>Abrahams, J.</dc:creator>
<dc:creator>Atrih, A.</dc:creator>
<dc:creator>Lamont, D. J.</dc:creator>
<dc:creator>Gierlinski, M.</dc:creator>
<dc:creator>Jayaprakash, P.</dc:creator>
<dc:creator>Michel, A. M.</dc:creator>
<dc:creator>Aboagye, E.</dc:creator>
<dc:creator>Yuneva, M.</dc:creator>
<dc:creator>Masson, G. R.</dc:creator>
<dc:creator>Shahrezaei, V.</dc:creator>
<dc:creator>Auner, H. W.</dc:creator>
<dc:date>2024-09-13</dc:date>
<dc:identifier>doi:10.1101/2024.09.10.611650</dc:identifier>
<dc:title><![CDATA[An integrated stress response-independent role of GCN2 prevents excessive ribosome biogenesis and mRNA translation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.09.611621v1?rss=1">
<title>
<![CDATA[
Mutations in PSEN1 predispose inflammation in an astrocyte model of familial Alzheimer's disease through disrupted regulated intramembrane proteolysis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.09.611621v1?rss=1"
</link>
<description><![CDATA[
Mutations in PSEN1 cause familial Alzheimers disease with almost complete penetrance. Age at onset is highly variable between different PSEN1 mutations and even within families with the same mutation. Current research into late onset Alzheimers disease implicates inflammation in both disease onset and progression. PSEN1 is the catalytic subunit of {gamma}-secretase, responsible for regulated intramembrane proteolysis of numerous substrates that include cytokine receptors. For this reason, we tested the hypothesis that mutations in PSEN1 impact inflammatory responses in astrocytes, thereby contributing to disease progression.

Here, using iPSC-astrocytes, we show that PSEN1 is upregulated in response to inflammatory stimuli, and this upregulation is disrupted by pathological PSEN1 mutations. Using transcriptomic analyses, we demonstrate that PSEN1 mutant astrocytes have an augmented inflammatory profile in their basal state, concomitant with an upregulation of genes coding for regulated intramembrane proteolytic and robust activation of JAK-STAT signalling. Using JAK-STAT2 as an example signalling pathway, we show altered phosphorylation cascades in PSEN1 mutant astrocytes, reinforcing the notion of altered cytokine signalling cascades. Finally, we use small molecule modulators of {gamma}-secretase to confirm a role for PSEN1/{gamma}-secretase in regulating the astrocytic response to inflammatory stimuli.

Together, these data suggest that mutations in PSEN1 enhance cytokine signalling via impaired regulated intramembrane proteolysis, thereby predisposing astrocytic inflammatory profiles. These findings support a two-hit contribution of PSEN1 mutations to fAD pathogenesis, not only impacting APP and A{beta} processing but also altering the cellular response to inflammation.
]]></description>
<dc:creator>Ziff, O. J.</dc:creator>
<dc:creator>Jolly, S.</dc:creator>
<dc:creator>Casey, J. M.</dc:creator>
<dc:creator>Granat, L.</dc:creator>
<dc:creator>Samra, S.</dc:creator>
<dc:creator>Seto-Salvia, N.</dc:creator>
<dc:creator>Alatza, A.</dc:creator>
<dc:creator>Phadke, L.</dc:creator>
<dc:creator>Galet, B.</dc:creator>
<dc:creator>Ravassard, P.</dc:creator>
<dc:creator>Potier, M.-C.</dc:creator>
<dc:creator>Fox, N. C.</dc:creator>
<dc:creator>Hardy, J.</dc:creator>
<dc:creator>Salih, D.</dc:creator>
<dc:creator>Whiting, P.</dc:creator>
<dc:creator>Ducotterd, F.</dc:creator>
<dc:creator>Patani, R.</dc:creator>
<dc:creator>Wray, S.</dc:creator>
<dc:creator>Arber, C.</dc:creator>
<dc:date>2024-09-13</dc:date>
<dc:identifier>doi:10.1101/2024.09.09.611621</dc:identifier>
<dc:title><![CDATA[Mutations in PSEN1 predispose inflammation in an astrocyte model of familial Alzheimer's disease through disrupted regulated intramembrane proteolysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.11.612412v1?rss=1">
<title>
<![CDATA[
SOX2 drives fetal reprogramming and reversible dormancy in colorectal cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.11.612412v1?rss=1"
</link>
<description><![CDATA[
Cellular plasticity plays critical roles in tissue regeneration, tumour progression and therapeutic resistance. However, the mechanism underlying this cell state transition remains elusive. Here, we show that the transcription factor SOX2 induces fetal reprogramming and reversible dormancy in colorectal cancer (CRC). SOX2 expression correlates with fetal reprogramming and poor prognosis in human primary and metastatic colorectal adenocarcinomas. In mouse CRC models, rare slow-cycling SOX2+SCA1+ cells are detected in early Apc-deleted mouse tumours that undergo slow clonal expansion over time. In contrast, the SOX2+ clones were found proliferative in advanced Apc-/-;KrasG12D/+;p53-/-;Tgfbr2-/-(AKPT) tumours, accompanied by dynamic cell state reprogramming from LGR5+ to LGR5-SCA1+ cells. Using transgenic mouse models, we demonstrate that ectopic expression of SOX2 inhibits intestinal lineage differentiation and induces fetal reprogramming. SOX2+ cells adopt dynamic cell cycle states depending on its expression level. High SOX2 expression results in hyperproliferation, whereas low SOX2 levels induce senescence-mediated dormancy. We find that loss of p53 can reverse SOX2-induced senescence, in line with the dormant cell state exit of the SOX2+ cells observed in advanced tumours. Finally, SOX2 expression is induced by 5-FU treatment in CRC. SOX2-expressing organoids exhibit increased tolerance to chemotherapy treatment, whilst deletion of SOX2 in AKPT tumour organoids sensitises drug responses. We propose that SOX2-induced plasticity and reversible dormancy promotes tumour progression and drug tolerance in CRC.
]]></description>
<dc:creator>Baulies, A.</dc:creator>
<dc:creator>Moncho-Amor, V.</dc:creator>
<dc:creator>Drago-Garcia, D.</dc:creator>
<dc:creator>Kucharska, A.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Moreno-Valladares, M.</dc:creator>
<dc:creator>Cruces-Salguero, S.</dc:creator>
<dc:creator>Hubl, F.</dc:creator>
<dc:creator>Hutton, C.</dc:creator>
<dc:creator>Kim, H.</dc:creator>
<dc:creator>Matheu, A.</dc:creator>
<dc:creator>Lovell-Badge, R.</dc:creator>
<dc:creator>Li, V. S. W.</dc:creator>
<dc:date>2024-09-16</dc:date>
<dc:identifier>doi:10.1101/2024.09.11.612412</dc:identifier>
<dc:title><![CDATA[SOX2 drives fetal reprogramming and reversible dormancy in colorectal cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.04.03.535433v1?rss=1">
<title>
<![CDATA[
Asparagine availability controls germinal centre B cell homeostasis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.04.03.535433v1?rss=1"
</link>
<description><![CDATA[
Germinal centre (GC) B cells proliferate at some of the highest rates of any mammalian cell, yet the metabolic processes which enable this are poorly understood. We performed integrated metabolomic and transcriptomic profiling of GC B cells, and found that metabolism of the non-essential amino acid asparagine (Asn) was highly upregulated. Asn was conditionally essential to B cells, and its synthetic enzyme, asparagine synthetase (ASNS) was upregulated following their activation, particularly more markedly in the absence of Asn, through the integrated stress response sensor general control non-derepressible 2 (GCN2). When Asns is deleted B cell survival and proliferation in low Asn conditions were strongly impaired, and removal of environmental Asn by asparaginase or dietary restriction markedly compromised the GC reaction, impairing affinity maturation and the humoral response to influenza infection. Using stable isotope tracing and single cell RNA sequencing, we found that metabolic adaptation to the absence of Asn requires ASNS, and that oxidative phosphorylation, mitochondrial homeostasis, and synthesis of nucleotides was particularly sensitive to Asn deprivation. Altogether, we reveal that Asn metabolism acts as a key regulator of B cell function and GC homeostasis.

The one sentence summaryAsparagine metabolism is a critical regulator of B cell function, maintaining the germinal centre reaction.
]]></description>
<dc:creator>Marin, E.</dc:creator>
<dc:creator>Bentkowska, K.</dc:creator>
<dc:creator>Yazicioglu, Y. F.</dc:creator>
<dc:creator>Clarke, A.</dc:creator>
<dc:date>2023-04-05</dc:date>
<dc:identifier>doi:10.1101/2023.04.03.535433</dc:identifier>
<dc:title><![CDATA[Asparagine availability controls germinal centre B cell homeostasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.15.613123v1?rss=1">
<title>
<![CDATA[
SIRPalpha+ PD-L1+ bone marrow macrophages aid AML growth by modulating T cell function 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.15.613123v1?rss=1"
</link>
<description><![CDATA[
Acute myeloid leukemia (AML) continues to have a poor prognosis due to its ability to relapse following initial response to chemotherapy. While immunotherapies hold the promise to revolutionize cancer treatment, AML has been particularly challenging to target. It is therefore important to better understand the relationship between AML cells and immune cells within the bone marrow (BM) microenvironment, where this disease grows. Here we focus on non-malignant BM macrophages, and using a combination of intravital microscopy, flow cytometry, transcriptomics and functional analyses we identify a subpopulation of immunomodulatory BM macrophages (IMMs) with a unique profile and function during AML progression. While the majority of macrophages are already being lost at early infiltration, IMMs are locally enriched. They are capable of efferocytosis and support AML growth through inhibition of T cells. Enrichment of IMMs in the BM of patients developing early relapse indicates that future development of interventions that target IMMs development and function may improve AML patients outcome.
]]></description>
<dc:creator>Birch, F.</dc:creator>
<dc:creator>Gonzalez-Anton, S.</dc:creator>
<dc:creator>Haltalli, M. L. R.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Bruno, F.</dc:creator>
<dc:creator>Georgiou, C.</dc:creator>
<dc:creator>Mai, C.</dc:creator>
<dc:creator>Todd, Y.</dc:creator>
<dc:creator>Payne, E.</dc:creator>
<dc:creator>Blum, S.</dc:creator>
<dc:creator>Pospori, C.</dc:creator>
<dc:creator>Arber, C.</dc:creator>
<dc:creator>Burt, R.</dc:creator>
<dc:creator>Tissot, F. S.</dc:creator>
<dc:creator>Celso, C. L.</dc:creator>
<dc:date>2024-09-19</dc:date>
<dc:identifier>doi:10.1101/2024.09.15.613123</dc:identifier>
<dc:title><![CDATA[SIRPalpha+ PD-L1+ bone marrow macrophages aid AML growth by modulating T cell function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.19.613857v1?rss=1">
<title>
<![CDATA[
The human ribosome modulates multidomain protein biogenesis by delaying cotranslational domain docking 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.19.613857v1?rss=1"
</link>
<description><![CDATA[
Proteins with multiple domains are intrinsically prone to misfold, yet fold efficiently during their synthesis on the ribosome. This is especially important in eukaryotes, where multidomain proteins predominate. Here, we sought to understand how multidomain protein folding is modulated by the eukaryotic ribosome. We used cryo-electron microscopy and hydrogen/deuterium exchange-mass spectrometry to characterise the structure and dynamics of partially-synthesised intermediates of a model multidomain protein. We find that nascent subdomains fold progressively during synthesis on the human ribosome, templated by interactions across domain interfaces. The conformational ensemble of the nascent chain is tuned by its unstructured C-terminal segments, which keep interfaces between folded domains in dynamic equilibrium until translation termination. This contrasts with the bacterial ribosome, on which domain interfaces form early and remain stable during synthesis. Delayed domain docking may avoid interdomain misfolding to promote the maturation of multidomain proteins in eukaryotes.
]]></description>
<dc:creator>Pellowe, G. A.</dc:creator>
<dc:creator>Voisin, T. B.</dc:creator>
<dc:creator>Karpauskaite, L.</dc:creator>
<dc:creator>Maslen, S. L.</dc:creator>
<dc:creator>Roeselova, A.</dc:creator>
<dc:creator>Skehel, M.</dc:creator>
<dc:creator>Roustan, C.</dc:creator>
<dc:creator>George, R.</dc:creator>
<dc:creator>Nans, A.</dc:creator>
<dc:creator>Kjaer, S.</dc:creator>
<dc:creator>Taylor, I. A.</dc:creator>
<dc:creator>Balchin, D.</dc:creator>
<dc:date>2024-09-19</dc:date>
<dc:identifier>doi:10.1101/2024.09.19.613857</dc:identifier>
<dc:title><![CDATA[The human ribosome modulates multidomain protein biogenesis by delaying cotranslational domain docking]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.20.614075v1?rss=1">
<title>
<![CDATA[
An intrinsically disordered RNA-binding region provides local target selectivity and is essential for LARP6 function. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.20.614075v1?rss=1"
</link>
<description><![CDATA[
Intrinsically disordered regions (IDRs) are prevalent in RNA-binding proteins (RBPs), yet their roles in RNA interactions remain poorly defined. Here, we examined the structured and disordered RNA-binding activities of LARP6, an RBP with a diverse RNA-binding repertoire. Using mass spectrometry-based RNA interaction mapping in living cells, we identified direct LARP6-RNA contacts within both the structured La-module and its flanking IDRs. Mutagenesis combined with individual-nucleotide resolution UV crosslinking and immunoprecipitation (iCLIP) revealed the La-module, but not the IDRs, as essential for LARP6 binding to RNA. Deletion of the N-terminal IDR broadened the footprints of LARP6 on RNA, uncovering a role in RNA-binding selectivity. Mechanistically, this is achieved through restricting the conformational flexibility of the adjacent La-module. The IDR-mediated RNA-binding selectivity is critical for LARP6-driven cancer cell viability and invasion. Our findings uncover a previously unrecognised critical function for IDRs in promoting selective RBP-RNA interactions, which operates through conformational restriction.
]]></description>
<dc:creator>Capraro, F.</dc:creator>
<dc:creator>Abis, G.</dc:creator>
<dc:creator>Karimzadeh, M.</dc:creator>
<dc:creator>Masino, L.</dc:creator>
<dc:creator>Barley, A.</dc:creator>
<dc:creator>Bui, T. T. T.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Conte, M. R.</dc:creator>
<dc:creator>Mardakheh, F. K.</dc:creator>
<dc:date>2024-09-20</dc:date>
<dc:identifier>doi:10.1101/2024.09.20.614075</dc:identifier>
<dc:title><![CDATA[An intrinsically disordered RNA-binding region provides local target selectivity and is essential for LARP6 function.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.22.613647v1?rss=1">
<title>
<![CDATA[
Divergent Plasmodium protein kinases drive MTOC, kinetochore and axoneme organisation in male gametogenesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.22.613647v1?rss=1"
</link>
<description><![CDATA[
Sexual development and male gamete formation of the malaria parasite in the mosquito midgut is governed by rapid endomitosis in the activated male gametocyte. This process is highly regulated by protein phosphorylation, specifically by three divergent male-specific protein kinases (PKs): CDPK4, SRPK1 and MAP2. Here, we localise each PK during rapid male gamete formation using live-cell imaging, identify their putative substrates by immunoprecipitation, and determine the morphological consequences of their absence using ultrastructure expansion and transmission electron microscopy. Each PK has a distinct location in either the nuclear or cytoplasmic compartment. Protein interaction studies revealed that CDPK4 and MAP2 interact with key drivers of rapid DNA replication, while SRPK1 is involved in RNA translation. The absence of each PK results in severe defects in either microtubule organising centre (MTOC) organisation, kinetochore segregation or axoneme formation. This study reveals the crucial role of these PKs during endomitosis in formation of the flagellated male gamete and uncovers some of their potential substrates that drive this process.

Summary blurbThis study reveals how Plasmodium kinases regulate MTOC, axoneme and kinetochore organisation in male gametogenesis, providing key insights into potential targets for malaria transmission control.
]]></description>
<dc:creator>Yanase, R.</dc:creator>
<dc:creator>Zeeshan, M.</dc:creator>
<dc:creator>Ferguson, D. J.</dc:creator>
<dc:creator>Markus, R.</dc:creator>
<dc:creator>Brady, D.</dc:creator>
<dc:creator>Bottrill, A. R.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:creator>Guttery, D. S.</dc:creator>
<dc:creator>Tewari, R.</dc:creator>
<dc:date>2024-09-22</dc:date>
<dc:identifier>doi:10.1101/2024.09.22.613647</dc:identifier>
<dc:title><![CDATA[Divergent Plasmodium protein kinases drive MTOC, kinetochore and axoneme organisation in male gametogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.23.614477v1?rss=1">
<title>
<![CDATA[
PDZ-directed substrate recruitment is the primary determinant of specific 4E-BP1dephosphorylation by PP1-Neurabin 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.23.614477v1?rss=1"
</link>
<description><![CDATA[
Protein Phosphatase 1 (PP1) relies on association with PP1-interacting proteins (PIPs) to generate substrate-specific PIP/PP1 holoenzymes, but the lack of well-defined substrates has hindered elucidation of the mechanisms involved. We previously demonstrated that the Phactr1 PIP confers sequence specificity on the Phactr1/PP1 holoenzyme by remodelling the PP1 hydrophobic substrate groove. Phactr1 defines a group of "RVxF-{Phi}{Phi}-R-W" PIPs that all interact with PP1 in a similar fashion. Here we use a PP1-PIP fusion approach to address sequence specificity and identify substrates of the RVxF-{Phi}{Phi}-R-W family PIPs. We show that the four Phactr proteins confer identical sequence specificities on their holoenzymes. We identify the 4E-BP and p70 S6K translational regulators as substrates for the Neurabin/Spinophilin PIPs, implicated in neuronal plasticity, pointing to a role for their holoenzymes in mTORC1-dependent translational control. Biochemical and structural experiments show that in contrast to the Phactrs, substrate recruitment and catalytic efficiency of the PP1-Neurabin and PP1-Spinophilin fusions is primarily determined by substrate interaction with the PDZ domain adjoining their RVxF-{Phi}{Phi}-R-W motifs, rather than by recognition of the remodelled PP1 hydrophobic groove. Thus, even PIPs that interact with PP1 in a similar manner use different mechanisms to ensure substrate selectivity.
]]></description>
<dc:creator>Fedoryshchak, R. O.</dc:creator>
<dc:creator>El-Bouri, K.</dc:creator>
<dc:creator>Joshi, D.</dc:creator>
<dc:creator>Mouilleron, S.</dc:creator>
<dc:creator>Treisman, R.</dc:creator>
<dc:date>2024-09-23</dc:date>
<dc:identifier>doi:10.1101/2024.09.23.614477</dc:identifier>
<dc:title><![CDATA[PDZ-directed substrate recruitment is the primary determinant of specific 4E-BP1dephosphorylation by PP1-Neurabin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.24.614722v1?rss=1">
<title>
<![CDATA[
A Drosophila holidic diet optimised for growthand development 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.24.614722v1?rss=1"
</link>
<description><![CDATA[
Diets composed of chemically pure components (holidic diets) are useful for determining the metabolic roles of individual nutrients. For the model organism Drosophila melanogaster, existing holidic diets are unable to support rapid growth characteristic of the larval stage. Here, we use a nutrient co-optimisation strategy across more than 50 diet variants to design HoldFast, a holidic medium tailored to fast larval growth and development. We identify dietary amino acid ratios optimal for developmental speed but show that they compromise survival unless vitamins and sterols are co-optimised. Rapid development on HoldFast is not improved by adding fatty acids but it is dependent upon their de novo synthesis in the fat body via fatty acid synthase (FASN). HoldFast outperforms other holidic diets, supporting rates of growth and development close to those of yeast-based diets and, under germ-free conditions, identical. HoldFast provides new opportunities for studying growth and metabolism during Drosophila development.
]]></description>
<dc:creator>Sorge, S.</dc:creator>
<dc:creator>Girard, V.</dc:creator>
<dc:creator>Lampe, L.</dc:creator>
<dc:creator>Tixier, V.</dc:creator>
<dc:creator>Weaver, A.</dc:creator>
<dc:creator>Higgins, T.</dc:creator>
<dc:creator>Gould, A. P.</dc:creator>
<dc:date>2024-09-24</dc:date>
<dc:identifier>doi:10.1101/2024.09.24.614722</dc:identifier>
<dc:title><![CDATA[A Drosophila holidic diet optimised for growthand development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.26.615224v1?rss=1">
<title>
<![CDATA[
Capture, mutual inhibition and release mechanism for aPKC-Par6 and its multi-site polarity substrate Lgl 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.26.615224v1?rss=1"
</link>
<description><![CDATA[
The mutually antagonistic kinase-substrate relationship between the apical aPKC-Par6 heterodimer and the basolateral substrate Lgl is key to the establishment and maintenance of cell polarity across metazoa. Although aPKC-Par6 can phosphorylate Lgl at three serine sites to exclude it from the apical domain, paradoxically, aPKC-Par6 and Lgl can also form a stable kinase-substrate complex whose function remains unclear and with conflicting roles proposed for Par6. We report the structure of human aPKC{iota}-Par6 bound to full-length Llgl1, captured through an aPKC{iota} docking site and a Par6PDZ contact. This soluble tripartite complex traps a phospho-S663 Llgl1 intermediate bridging between aPKC and Par6, impeding phosphorylation progression. Thus, aPKC{iota} is effectively inhibited by Llgl1pS663 whilst Llgl1 is captured by aPKC{iota}-Par6. Mutational disruption of Lgl-aPKC interaction impedes complex assembly and Lgl phosphorylation, whereas disrupting the Lgl-Par6PDZ contact promotes complex dissociation and completion of Lgl phosphorylation cycle. We incorporate these findings into a Par6PDZ-regulated substrate capture-and-release model that we demonstrate requires binding by Cdc42-GTP and the apical partner Crumbs to drive complex disassembly. Our results provide an explanation for the opposing roles of Par6 underpinning the spatial control of aPKC-Par6 activity by Lgl relevant to polarised membrane contexts across multi-cellular organisms.
]]></description>
<dc:creator>Earl, C. P.</dc:creator>
<dc:creator>Cobbaut, M.</dc:creator>
<dc:creator>Carvalho, A. B.</dc:creator>
<dc:creator>Ivanova, M. E.</dc:creator>
<dc:creator>Briggs, D. C.</dc:creator>
<dc:creator>Morais-de-Sa, E.</dc:creator>
<dc:creator>Parker, P. J.</dc:creator>
<dc:creator>McDonald, N. Q.</dc:creator>
<dc:date>2024-09-26</dc:date>
<dc:identifier>doi:10.1101/2024.09.26.615224</dc:identifier>
<dc:title><![CDATA[Capture, mutual inhibition and release mechanism for aPKC-Par6 and its multi-site polarity substrate Lgl]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.26.614906v1?rss=1">
<title>
<![CDATA[
Live imaging human embryos reveals mitotic errors and lineage specification prior to implantation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.26.614906v1?rss=1"
</link>
<description><![CDATA[
Meiotic and mitotic chromosome segregation errors in human development have been studied mostly prior to and at the time of fertilisation. Despite chromosomal errors being a leading cause of miscarriage and infertility, chromosome missegregation has not been extensively studied at later stages of human development. Here we optimised labelling, light-sheet live imaging and semi-quantitative analysis of human embryos and reveal chromosome segregation errors just prior to implantation. We found that human embryos exhibited a number of chromosome missegregation events including multipolar spindle formation, lagging chromosomes, misalignment and chromosome slippage. We found that the majority of lagging chromosomes were passively inherited by one of the daughter cells, instead of reincorporating into the nuclei, suggesting a distinct pattern of micronuclei inheritance. By semi-automated segmentation, we tracked the position of labelled cells in human embryos and observed that while most labelled cells remained segregated to the outside, and therefore restricted to a placental-progenitor fate, there was evidence of a rare cell migration to cells positioned on the inside, which suggests that there may be plasticity. Altogether, we found that mitotic chromosome segregation errors arise just prior to implantation, which has implications for our understanding of biological events that contribute to aneuploidy mosaicism.
]]></description>
<dc:creator>Abdelaal, A. A.</dc:creator>
<dc:creator>Mccarthy, A.</dc:creator>
<dc:creator>Karsa, A.</dc:creator>
<dc:creator>Muresan, L.</dc:creator>
<dc:creator>Elder, K.</dc:creator>
<dc:creator>Papathanasiou, A.</dc:creator>
<dc:creator>Snell, P.</dc:creator>
<dc:creator>Christie, L.</dc:creator>
<dc:creator>Wilding, M.</dc:creator>
<dc:creator>Steventon, B.</dc:creator>
<dc:creator>Niakan, K.</dc:creator>
<dc:date>2024-09-26</dc:date>
<dc:identifier>doi:10.1101/2024.09.26.614906</dc:identifier>
<dc:title><![CDATA[Live imaging human embryos reveals mitotic errors and lineage specification prior to implantation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.25.614952v1?rss=1">
<title>
<![CDATA[
Differential responses and recovery dynamics of HSPC populations following Plasmodium chabaudi infection. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.25.614952v1?rss=1"
</link>
<description><![CDATA[
Severe infections such as malaria are on the rise worldwide, driven by both climate change and increasing drug-resistance. It is therefore paramount that we better understand how the hosts organism responds to severe infection. Hematopoiesis is particularly of interest in this context because hematopoietic stem and progenitor cells (HSPCs) maintain the turnover of all blood cells, including all immune cells. Severe infections have been widely acknowledged to affecting HSPCs, however this disruption has been mainly studied during the acute phase, and the process and level of HSPC recovery remains understudied. Using a self-resolving model of natural rodent malaria, infection by Plasmodium chabaudi, here we systematically assess HSPCs acute response and recovery upon pathogen clearance. We demonstrate that during the acute phase of infection the most quiescent and functional stem cells are depleted, multipotent progenitor compartments are drastically enlarged, and oligopotent progenitors virtually disappear, underpinned by dramatic, population-specific and sometimes unexpected changes in proliferation rates. HSPC populations return to homeostatic size and proliferation rate again through specific patterns of recovery. Overall, our data demonstrate that HSPC populations adopt different responses to cope with severe infection and suggest that the ability to adjust proliferative capacity becomes more restricted as differentiation progresses.
]]></description>
<dc:creator>Bruno, F.</dc:creator>
<dc:creator>Georgiou, C.</dc:creator>
<dc:creator>Cunningham, D.</dc:creator>
<dc:creator>Atkinson, S.</dc:creator>
<dc:creator>Bett, L.</dc:creator>
<dc:creator>Secchi, M.</dc:creator>
<dc:creator>Birch, F.</dc:creator>
<dc:creator>Gonzalez Anton, S.</dc:creator>
<dc:creator>langhorne, j.</dc:creator>
<dc:creator>Lo Celso, C. L.</dc:creator>
<dc:date>2024-09-27</dc:date>
<dc:identifier>doi:10.1101/2024.09.25.614952</dc:identifier>
<dc:title><![CDATA[Differential responses and recovery dynamics of HSPC populations following Plasmodium chabaudi infection.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.27.615480v1?rss=1">
<title>
<![CDATA[
Mycobacterium tuberculosis overcomes phosphate starvation by extensively remodelling its lipidome with phosphorus-free lipids 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.27.615480v1?rss=1"
</link>
<description><![CDATA[
Tuberculosis (TB) is the biggest cause of death from infectious disease worldwide [1-4]. The causative agent, Mycobacterium tuberculosis (Mtb), possesses a complex cell envelope that determines many of the key physiologic and virulence properties that facilitate infection, which comprises multiple classes of unique lipids [5-7]. The macrophage phagosome is a key reservoir of infection in pulmonary TB and multiple studies have shown that inorganic phosphate (Pi) is limiting in this environment [8-11]. The ability of Mtb to sense and respond to phosphate starvation is required for virulence in animal models and replication in human macrophages in vitro [12-14]. Here, we show that during Pi restriction the Mtb lipidome is markedly remodelled such that phospholipids are replaced with multiple classes of phosphorus-free lipids, some of which have not been documented before. Further, we discover that Mtb can metabolise phospholipid polar heads derived from host pulmonary surfactant as an alternative phosphate source, which sustains cell division for several generations during Pi restriction. These dual manipulations of phospholipid metabolism provide Mtb with an escape from phosphate restriction specific to the infection of alveolar macrophages, one of the earliest steps in establishing pulmonary tuberculosis. The changes in envelope lipidome remodelling, akin to that observed in some marine and terrestrial bacteria [15-20] suggests that standard Mtb culture conditions that use media with high concentrations of Pi do not reflect the physiologic environment during infection, thereby potentially undermining vaccine and drug development for tuberculosis. Moreover, the distinct Mtb phosphate-free lipids and the metabolic pathways that generate them could provide new antibiotic targets.
]]></description>
<dc:creator>Gray, R. M.</dc:creator>
<dc:creator>Hunt, D. M.</dc:creator>
<dc:creator>Silva dos Santos, M.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Agapova, A.</dc:creator>
<dc:creator>Rodgers, A.</dc:creator>
<dc:creator>Garza-Garcia, A.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Gutierrez, M. G.</dc:creator>
<dc:creator>Lee, R.</dc:creator>
<dc:creator>Carvalho, L. P.</dc:creator>
<dc:date>2024-09-27</dc:date>
<dc:identifier>doi:10.1101/2024.09.27.615480</dc:identifier>
<dc:title><![CDATA[Mycobacterium tuberculosis overcomes phosphate starvation by extensively remodelling its lipidome with phosphorus-free lipids]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.27.615442v1?rss=1">
<title>
<![CDATA[
A depth map of visual space in the primary visual cortex 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.27.615442v1?rss=1"
</link>
<description><![CDATA[
Depth perception is essential for visually-guided behavior. Computer vision algorithms use depth maps to encode distances in three-dimensional scenes but it is unknown whether such depth maps are generated by animal visual systems. To answer this question, we focused on motion parallax, a depth cue relying on visual motion resulting from movement of the observer. As neurons in the mouse primary visual cortex (V1) are broadly modulated by locomotion, we hypothesized that they may integrate vision- and locomotion-related signals to estimate depth from motion parallax. Using recordings in a three-dimensional virtual reality environment, we found that conjunctive coding of visual and self-motion speeds gave rise to depth-selective neuronal responses. Depth-selective neurons could be characterized by three-dimensional receptive fields, responding to specific virtual depths and retinotopic locations. Neurons tuned to a broad range of virtual depths were found across all sampled retinotopic locations, showing that motion parallax generates a depth map of visual space in V1.
]]></description>
<dc:creator>He, Y.</dc:creator>
<dc:creator>Colas Nieto, A.</dc:creator>
<dc:creator>Blot, A.</dc:creator>
<dc:creator>Znamenskiy, P.</dc:creator>
<dc:date>2024-09-30</dc:date>
<dc:identifier>doi:10.1101/2024.09.27.615442</dc:identifier>
<dc:title><![CDATA[A depth map of visual space in the primary visual cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.01.616014v1?rss=1">
<title>
<![CDATA[
A stable Netrin-1 fluorescent reporter chicken reveals cell-specific molecular signaturesduring optic fissure closure. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.01.616014v1?rss=1"
</link>
<description><![CDATA[
NTN1 is expressed in a wide range of developmental tissues and is essential for normal development. Here we describe the generation of a Netrin-1 reporter chicken line (NTN1-T2A-eGFP) by targeting green fluorescent protein into the NTN1 locus using CRISPR/Cas9 methodology. Our strategy gave 100% transmission of heterozygous (NTN1T2A-eGFP/+) embryos in which GFP localisation faithfully replicated endogenous NTN1 expression in the optic fissure and neural tube floorplate. Furthermore, all NTN1T2A-eGFP/+ embryos and hatched birds appeared phenotypically normal. We applied this resource to a pertinent developmental context - coloboma is a structural eye malformation characterised by failure of epithelial fusion during optic fissure closure (OFC) and NTN1 is specifically expressed in fusion pioneer cells at the edges of the optic fissure. We therefore optimised the isolation of GFP expressing cells from embryonic NTN1T2A-eGFP/+ eyes using spectral fluorescence cell-sorting and applied transcriptomic profiling of pioneer cells, which revealed multiple new OFC markers and novel pathways for developmental tissue fusion and coloboma. This work provides a novel fluorescent NTN1 chicken reporter line with broad experimental utility and is the first to directly molecularly characterise pioneer cells during OFC.
]]></description>
<dc:creator>Chan, B. H. C.</dc:creator>
<dc:creator>Hardy, H.</dc:creator>
<dc:creator>Requena, T.</dc:creator>
<dc:creator>Findlay, A.</dc:creator>
<dc:creator>Ioannidis, J.</dc:creator>
<dc:creator>Meunier, D.</dc:creator>
<dc:creator>Toms, M.</dc:creator>
<dc:creator>Moosajee, M.</dc:creator>
<dc:creator>Raper, A.</dc:creator>
<dc:creator>McGrew, M.</dc:creator>
<dc:creator>Rainger, J.</dc:creator>
<dc:date>2024-10-02</dc:date>
<dc:identifier>doi:10.1101/2024.10.01.616014</dc:identifier>
<dc:title><![CDATA[A stable Netrin-1 fluorescent reporter chicken reveals cell-specific molecular signaturesduring optic fissure closure.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.01.616065v1?rss=1">
<title>
<![CDATA[
Meru co-ordinates spindle orientation with cell polarity and cell cycle progression 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.01.616065v1?rss=1"
</link>
<description><![CDATA[
Correct mitotic spindle alignment is essential for tissue architecture and plays an important role in cell fate specification through asymmetric cell division. Spindle tethering factors such as Drosophila Mud (NuMA in mammals) are recruited to the cell cortex and capture astral microtubules, pulling the spindle in the correct orientation. However, how spindle tethering complexes read the cell polarity axis and how spindle attachment is coupled to mitotic progression remains poorly understood. We explore these questions in Drosophila sensory organ precursors (SOPs), which divide asymmetrically to give rise to the epidermal mechanosensory bristles. We show that the scaffold protein Meru, which is recruited to the posterior cortex by the Frizzled/Dishevelled planar cell polarity complex, in turn recruits Mud, linking the spindle tethering and polarity machineries. Furthermore, Cyclin A/Cdk1 associates with Meru at the posterior cortex, promoting the formation of the Mud/Meru/Dsh complex via Meru and Dsh phosphorylation. Thus, Meru couples spindle orientation with cell polarity and provides a cell cycle-dependent cue for spindle tethering.
]]></description>
<dc:creator>McLellan, M. M.</dc:creator>
<dc:creator>Aerne, B. L.</dc:creator>
<dc:creator>Dhoul, J. J. B.</dc:creator>
<dc:creator>Tapon, N.</dc:creator>
<dc:date>2024-10-02</dc:date>
<dc:identifier>doi:10.1101/2024.10.01.616065</dc:identifier>
<dc:title><![CDATA[Meru co-ordinates spindle orientation with cell polarity and cell cycle progression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.02.616326v1?rss=1">
<title>
<![CDATA[
Growth compensation upon changes in tissue size in the Drosophila abdomen 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.02.616326v1?rss=1"
</link>
<description><![CDATA[
Attaining the appropriate size during development is essential for the function of animal tissues and organs. Robust tissue size control implies the existence of compensatory mechanisms that allow developing systems to recover from growth perturbations. However, the difficulty of directly observing normal or compensatory developmental growth means we have little understanding of the cellular behaviours that confer robustness to tissue size control. Here, we study how growth perturbations affect proliferation kinetics and the timing of growth termination of Drosophila histoblasts, the progenitors that give rise to the adult abdominal epidermis. Histoblasts undergo extensive growth and proliferation during the pupal stage, which is accessible for long-term live-imaging and precise quantitative analysis. By manipulating cell number or volume prior to the pupal growth phase, we changed the starting size of the abdomen primordium, then observed how the histoblasts adapted to these changes by altering their growth dynamics. We show that, upon a decrease in starting tissue size, the histoblasts compensate by extending their temporal proliferative window, undergoing additional cell cycles, as well as increasing their apical area to maximise coverage of the abdominal surface. When initial tissue size is increased, the histoblasts undergo fewer division cycles and arrest proliferation earlier than normal. Thus, the proliferative window of this tissue is flexible enough to buffer for changes in tissue size. Our data also suggest that the histoblasts sense both spatial and temporal cues to arrest their growth at the appropriate time and ensure accurate tissue size control.
]]></description>
<dc:creator>Ferreira, A.</dc:creator>
<dc:creator>Cairoli, A.</dc:creator>
<dc:creator>Mangione, F.</dc:creator>
<dc:creator>Holder, M. V.</dc:creator>
<dc:creator>Ainslie, A.</dc:creator>
<dc:creator>Aerne, B. L.</dc:creator>
<dc:creator>Salbreux, G.</dc:creator>
<dc:creator>Tapon, N.</dc:creator>
<dc:date>2024-10-03</dc:date>
<dc:identifier>doi:10.1101/2024.10.02.616326</dc:identifier>
<dc:title><![CDATA[Growth compensation upon changes in tissue size in the Drosophila abdomen]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.03.616443v1?rss=1">
<title>
<![CDATA[
DYRK1A kinase triplication is the major cause of Otitis Media in Down Syndrome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.03.616443v1?rss=1"
</link>
<description><![CDATA[
Down syndrome (DS), which arises from trisomy of the whole or part of chromosome 21 (Hsa21), is one of the most common genetic abnormalities in humans. DS manifests as a broad spectrum of phenotypic features, including hearing loss due to otitis media with effusion (OME), affecting around 50% of children with DS. We employed a panel of mouse models of DS comprising a nested series of duplications covering the regions of the mouse genome syntenic to Hsa21 in order to define the loci involved with OME in DS. We identified a major locus on mouse chromosome 16, containing only 12 genes, that causes OME. Within this region we demonstrate that normalizing the gene dosage of Dyrk1a restored the wild-type phenotype. Investigation of downstream pathways of DYRK1A uncovered a number of pathological mechanisms whereby DYRK1A triplication leads to middle ear inflammation and vascular leak. These include cross-talk of DYRK1A and TGF{beta} signaling and its impact on proinflammatory cytokines IL-6 and IL-17, as well as raised VEGF levels in the middle ear accompanied by increased Hif1a. We conclude that DYRK1A is a potential therapeutic target for OME in children with DS.
]]></description>
<dc:creator>Tateossian, H.</dc:creator>
<dc:creator>Southern, A.</dc:creator>
<dc:creator>Vikhe, P.</dc:creator>
<dc:creator>Lana-Elola, E.</dc:creator>
<dc:creator>Watson-Scales, S.</dc:creator>
<dc:creator>Gibbins, D.</dc:creator>
<dc:creator>Williams, D.</dc:creator>
<dc:creator>Purnell, T.</dc:creator>
<dc:creator>Mburu, P.</dc:creator>
<dc:creator>Parker, A.</dc:creator>
<dc:creator>Norris, D. P.</dc:creator>
<dc:creator>Santos-Cortez, R. L. P.</dc:creator>
<dc:creator>Herrmann, B. W.</dc:creator>
<dc:creator>Wells, S.</dc:creator>
<dc:creator>Lad, H. V.</dc:creator>
<dc:creator>Fisher, E.</dc:creator>
<dc:creator>Tybulewicz, V. L.</dc:creator>
<dc:creator>Brown, S. D. M.</dc:creator>
<dc:date>2024-10-03</dc:date>
<dc:identifier>doi:10.1101/2024.10.03.616443</dc:identifier>
<dc:title><![CDATA[DYRK1A kinase triplication is the major cause of Otitis Media in Down Syndrome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.07.616970v1?rss=1">
<title>
<![CDATA[
Presynaptic filopodia form kinapses and modulate membrane mechanics for synchronous neurotransmission and seizure generation. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.07.616970v1?rss=1"
</link>
<description><![CDATA[
The structural stability of synapses directly contrasts with their functional plasticity. This conceptual dichotomy is explained by the assumption that all synaptic plasticity is generated via either electrical and/or biochemical signaling. Here, we challenge this dogma by revealing an activity-dependent presynaptic response that is physical in nature. We show that dynamic filopodia emerge during action potential discharge and transiently deform synaptic boutons to enhance connectivity. Filopodia generation requires neuronal activity, calcium and actin, and occurs in intact brain circuits and human brain. Mechanistically, their extension preserves synchronous neurotransmitter release by increasing presynaptic membrane tension. However, filopodia generation becomes maladaptive during dysregulated brain activity, exacerbating seizures in vivo. Therefore, we provide direct evidence that presynaptic mechanical forces determine the extent and timing of synaptic signals.
]]></description>
<dc:creator>Jain, A.</dc:creator>
<dc:creator>Kroll, J.</dc:creator>
<dc:creator>Webster, J. F.</dc:creator>
<dc:creator>Moss, J.</dc:creator>
<dc:creator>Ultanir, S. K.</dc:creator>
<dc:creator>Gonzalez-Sulser, A.</dc:creator>
<dc:creator>Rosenmund, C.</dc:creator>
<dc:creator>Cousin, M. A.</dc:creator>
<dc:creator>Ivanova, D.</dc:creator>
<dc:date>2024-10-07</dc:date>
<dc:identifier>doi:10.1101/2024.10.07.616970</dc:identifier>
<dc:title><![CDATA[Presynaptic filopodia form kinapses and modulate membrane mechanics for synchronous neurotransmission and seizure generation.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.04.616491v1?rss=1">
<title>
<![CDATA[
Mapping the niche of breast cancer metastases in lung and liver 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.04.616491v1?rss=1"
</link>
<description><![CDATA[
Breast cancer progression to visceral organs such as lung and liver is regarded as a dreadful event, unequivocally associated with a poor prognosis. Yet, these vital sites are characterized by highly diverse cellular microenvironments and physiological functions, suggesting that they may influence cancer cells behavior in divergent ways. Unexpectedly, we find that while the liver microenvironment fosters metastasis-promoting properties and boosts secondary spread, the lungs impose a roadblock to the same processes. Using patient data and tissues from rapid autopsy, as well as mouse models with barcode-mediated metastasis tracing, niche labeling technology and single cell analysis of both tumor cells and their direct microenvironment, we dissect cellular and molecular microenvironmental factors that impose this differential behavior. Among these, we identify BMP2-producing endothelial cells as critical players within the liver metastatic niche, capable to enhance metastasis-to-metastasis dissemination. Targeting BMP2 receptor on breast cancer cells suppresses their metastasis-forming ability. Altogether, we reveal a contrast in the site-specific behavior of lung and liver metastases in breast cancer, highlighting microenvironmental factors that contribute to this diversity, as well as organ-specific opportunities for intervention.
]]></description>
<dc:creator>Sznurkowska, M. K.</dc:creator>
<dc:creator>Castro-Giner, F.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Ring, A.</dc:creator>
<dc:creator>Schwab, F. D.</dc:creator>
<dc:creator>Albrecht, F.</dc:creator>
<dc:creator>Mertz, K. D.</dc:creator>
<dc:creator>Strittmatter, K.</dc:creator>
<dc:creator>Ozimski, L. L.</dc:creator>
<dc:creator>Budinjas, S.</dc:creator>
<dc:creator>Auray, G.</dc:creator>
<dc:creator>Giachino, C.</dc:creator>
<dc:creator>Taylor, V.</dc:creator>
<dc:creator>Gvozdenovic, A.</dc:creator>
<dc:creator>Malanchi, I.</dc:creator>
<dc:creator>Frauchiger-Heuer, H.</dc:creator>
<dc:creator>Wicki, A.</dc:creator>
<dc:creator>Vetter, M.</dc:creator>
<dc:creator>Aceto, N.</dc:creator>
<dc:date>2024-10-07</dc:date>
<dc:identifier>doi:10.1101/2024.10.04.616491</dc:identifier>
<dc:title><![CDATA[Mapping the niche of breast cancer metastases in lung and liver]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.01.616066v1?rss=1">
<title>
<![CDATA[
The PHD3-FOXO3 axis modulates the interferon type I response in microglia aggravating Alzheimer's disease progression 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.01.616066v1?rss=1"
</link>
<description><![CDATA[
Microglia respond to Alzheimers disease (AD) with a variety of transcriptional responses. However, the regulation of specific transcriptional signatures and the contribution of each individual response to disease progression is only starting to be characterized. We have previously shown that hypoxia via hypoxia inducible factor 1 (HIF1) is a strong regulator of A{beta} plaque-associated microglia (A{beta}AM). Here, we characterize the role of HIF1-mediated transcription of Egln3, encoding for PHD3, in A{beta}AM. We show that oligomeric A{beta} treatment (oA{beta}) in vitro induces the expression of Hif1a and Egln3 in microglia, which correlates with the transcriptional activation of genes involved in the interferon type I signature (IFNS) in a PHD3-dependent manner. Mechanistically, we demonstrate FOXO3 to be an important repressor of IFNS in microglia, whose abundance decreases upon A{beta} presence, and, correspondingly, both in human single-nucleus (sn) and mouse A{beta}AM transcriptomics, FOXO3 DNA binding sites define the IFNS. FOXO3 repression of the IFNS is dependent on PHD3, with our results suggesting a physical interaction between both proteins in vitro. In vivo, loss of PHD3 correlate with abrogation of the IFNS and activation of the disease-associated microglia signature (DAM) in A{beta}AM. Transcriptional changes in microglia associate with increased microglia proximity to A{beta} plaques, augmented phagocytosis of A{beta} by microglia, reduced parenchymal levels of A{beta}, and an increase in small-sized plaques. PHD3 deficiency also reduced the A{beta} plaque-associated neuropathology and rescued behavioural deficits of an AD mouse model. Finally, we also demonstrate that microglial PHD3 overexpression during development in the absence of A{beta} pathology is sufficient to induce the IFNS and to behavioural alterations. Altogether, our data strongly indicate that the PHD3-FOXO3 axis controls the microglial IFNS in a cell autonomous manner, contributing to the progression of AD.
]]></description>
<dc:creator>Sanchez-Garcia, M. A.</dc:creator>
<dc:creator>Lara-Urena, N.</dc:creator>
<dc:creator>March-Diaz, R.</dc:creator>
<dc:creator>Ortega-de San Luis, C.</dc:creator>
<dc:creator>Quinones-Canete, S.</dc:creator>
<dc:creator>Barba-Reyes, J. M.</dc:creator>
<dc:creator>Cabello-Rivera, D.</dc:creator>
<dc:creator>Munoz-Cabello, A. M.</dc:creator>
<dc:creator>Mora-Romero, B.</dc:creator>
<dc:creator>Romero-Molina, C.</dc:creator>
<dc:creator>Heras-Garvin, A.</dc:creator>
<dc:creator>Navarro, V.</dc:creator>
<dc:creator>Lopez-Barneo, J.</dc:creator>
<dc:creator>Vizuete, M.</dc:creator>
<dc:creator>Vitorica, J.</dc:creator>
<dc:creator>Munoz-Machado, A. B.</dc:creator>
<dc:creator>Cockman, M.</dc:creator>
<dc:creator>Rosales-Nieves, A. E.</dc:creator>
<dc:creator>Pascual, A.</dc:creator>
<dc:date>2024-10-03</dc:date>
<dc:identifier>doi:10.1101/2024.10.01.616066</dc:identifier>
<dc:title><![CDATA[The PHD3-FOXO3 axis modulates the interferon type I response in microglia aggravating Alzheimer's disease progression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.15.614675v1?rss=1">
<title>
<![CDATA[
Identification of RING E3 pseudoligases in the TRIM protein family 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.15.614675v1?rss=1"
</link>
<description><![CDATA[
TRIpartite Motif (TRIM) family proteins have diverse roles across a broad variety of cellular functions, which are largely presumed to depend on their ubiquitin E3 ligase activity, conferred by a RING domain. However, recent reports have shown that some TRIMs lack detectable ubiquitination activity in isolation, despite containing a RING domain. Here, we present parallel in cellulo, in vitro, and in silico structure-function analyses of the ubiquitin E3 ligase activity and RING domain structural characteristics of whole TRIM protein family. In-depth follow-up studies of this comprehensive dataset reveals a number of  pseudoligases, whose RING domains have structurally diverged at either the homodimerisation or E2[~]ubiquitin interfaces, thereby disrupting their ability to catalyse ubiquitin transfer. Together, these data raise intriguing open questions regarding the unknown TRIM functions in physiology and disease.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=83 SRC="FIGDIR/small/614675v1_ufig1.gif" ALT="Figure 1">
View larger version (17K):
org.highwire.dtl.DTLVardef@7c851org.highwire.dtl.DTLVardef@f06afdorg.highwire.dtl.DTLVardef@3cd7org.highwire.dtl.DTLVardef@19a60e6_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Dudley-Fraser, J.</dc:creator>
<dc:creator>Esposito, D.</dc:creator>
<dc:creator>McPhie, K.</dc:creator>
<dc:creator>Morley-Williams, C.</dc:creator>
<dc:creator>Auchynnikava, T.</dc:creator>
<dc:creator>Rittinger, K.</dc:creator>
<dc:date>2024-10-17</dc:date>
<dc:identifier>doi:10.1101/2024.10.15.614675</dc:identifier>
<dc:title><![CDATA[Identification of RING E3 pseudoligases in the TRIM protein family]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.11.617838v1?rss=1">
<title>
<![CDATA[
Identification of a specialized lipid barrier for Drosophila metamorphosis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.11.617838v1?rss=1"
</link>
<description><![CDATA[
In many terrestrial insects, the onset of metamorphosis marks a transition from humid to dry environments. Yet how metamorphosing insect pupae protect themselves against the threat of dehydration remains unclear. Here, we identify the chemical composition and biosynthetic origins of a lipid desiccation barrier specific to the pupal and sexually-immature adult stages of Drosophila melanogaster. This barrier comprises unisex hyper-long hydrocarbons, 29-37 carbons in length, which are synthesized by larval oenocytes and stored in the larval fat body before being deployed on the pupal and young adult cuticles. We show that the fatty acid elongase EloHL is required for the biosynthesis of hyper-long hydrocarbons that are essential for the barrier to water loss during metamorphosis. Across the Drosophila genus, many species express unisex profiles of hyper-long hydrocarbons and, as young adults, transition to sex-specific shorter hydrocarbons with known pheromonal functions. The desert species D. mojavensis, however, retains hyper-long hydrocarbons during adulthood likely as an adaptation to an arid environment. Our study reveals how the cuticular lipid barrier is tuned to meet changing environmental pressures during insect development and evolution.
]]></description>
<dc:creator>Lampe, L.</dc:creator>
<dc:creator>Newell, C. L.</dc:creator>
<dc:creator>Wang, B.-J.</dc:creator>
<dc:creator>Makki, R.</dc:creator>
<dc:creator>Alexandre, C.</dc:creator>
<dc:creator>Gilmore, I. S.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:creator>Gould, A. P.</dc:creator>
<dc:date>2024-10-12</dc:date>
<dc:identifier>doi:10.1101/2024.10.11.617838</dc:identifier>
<dc:title><![CDATA[Identification of a specialized lipid barrier for Drosophila metamorphosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.14.618182v1?rss=1">
<title>
<![CDATA[
Integrative analysis of higher-order transcriptome organisation and RNA condensation principles 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.14.618182v1?rss=1"
</link>
<description><![CDATA[
Complex RNA-protein networks play a pivotal role in the formation of many types of biomolecular condensates. How intrinsic RNA features contribute to condensate formation however remains unclear. Here, we integrate tailored transcriptomics assays to identify a distinct class of developmental condensation-prone RNAs termed  smOOPs (semi-extractable, orthogonal organic phase separation-enriched RNAs). These transcripts are localised to larger intracellular foci, form denser RNA-RNA interaction subnetworks than expected and are heavily bound by RNA binding proteins (RBPs). Using an explainable deep learning framework, we reveal that smOOPs harbor characteristic sequence composition with lower sequence complexity, increased intramolecular folding and specific RBP binding patterns. Intriguingly, these RNAs encode proteins bearing extensive intrinsically disordered regions and are markedly predicted to be involved in biomolecular condensates, indicating an interplay between RNA- and protein-based features in phase separation. This work advances our understanding of condensation-prone RNAs and provides a versatile resource to further investigate RNA-driven condensation principles.
]]></description>
<dc:creator>Klobucar, T.</dc:creator>
<dc:creator>Novljan, J.</dc:creator>
<dc:creator>Iosub, I. A.</dc:creator>
<dc:creator>Kokot, B.</dc:creator>
<dc:creator>Urbancic, I.</dc:creator>
<dc:creator>Jones, D. M.</dc:creator>
<dc:creator>Chakrabarti, A. M.</dc:creator>
<dc:creator>Luscombe, N.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Modic, M.</dc:creator>
<dc:date>2024-10-16</dc:date>
<dc:identifier>doi:10.1101/2024.10.14.618182</dc:identifier>
<dc:title><![CDATA[Integrative analysis of higher-order transcriptome organisation and RNA condensation principles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.17.618945v1?rss=1">
<title>
<![CDATA[
Deletions Rate-Limit Breast and Ovarian Cancer Initiation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.17.618945v1?rss=1"
</link>
<description><![CDATA[
Optimizing prevention and early detection of cancer requires understanding the number, types and timing of driver mutations. To quantify this, we exploited the elevated cancer incidence and mutation rates in germline BRCA1 and BRCA2 (gBRCA1/2) carriers. Using novel statistical models, we identify genomic deletions as the likely rate-limiting mutational processes, with 1-3 deletions required to initiate breast and ovarian tumors. gBRCA1/2-driven hereditary and sporadic tumors undergo convergent evolution to develop a similar set of driver deletions, and deletions explain the elevated cancer risk of gBRCA1/2-carriers. Orthogonal mutation timing analysis identifies deletions of chromosome 17 and 13q as early, recurrent events. Single-cell analyses confirmed deletion rate differences in gBRCA1/2 vs. non-carrier tumors as well as cells engineered to harbor gBRCA1/2. The centrality of deletion-associated chromosomal instability to tumorigenesis shapes interpretation of the somatic evolution of non-malignant tissue and guides strategies for precision prevention and early detection.
]]></description>
<dc:creator>Houlahan, K.</dc:creator>
<dc:creator>Bihie, M.</dc:creator>
<dc:creator>Contreras, J. G.</dc:creator>
<dc:creator>Fulop, D.</dc:creator>
<dc:creator>Lopez, G.</dc:creator>
<dc:creator>Huang, H.-H.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>Curtis, C.</dc:creator>
<dc:creator>Boutros, P.</dc:creator>
<dc:creator>Huang, K.-l.</dc:creator>
<dc:date>2024-10-21</dc:date>
<dc:identifier>doi:10.1101/2024.10.17.618945</dc:identifier>
<dc:title><![CDATA[Deletions Rate-Limit Breast and Ovarian Cancer Initiation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.22.619613v1?rss=1">
<title>
<![CDATA[
A distinctive PI(4,5)P2 compartment forms during entosis and related engulfment processes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.22.619613v1?rss=1"
</link>
<description><![CDATA[
Entosis is a form of cell cannibalism prevalent in human tumours. During entosis, one live and viable epithelial cell is completely internalised by another, then housed inside a large, single-membrane, endolysosomal vacuole in the host cytosol. The composition and maturation of this specialised, macroendocytic compartment has not been fully defined, but ultimately, the inner cell is killed and digested by host lysosomes. Here, we investigate the molecular characteristics and maturation profile of the entotic vacuole. Like phagosomes and macropinosomes, this vacuole undergoes a series of phospholipid modifications, but its maturation profile bears distinctive dynamics. While PI(4,5)P2 is lost rapidly during phagosome maturation, entosis yields an unusual, intracellular PI(4,5)P2-positive compartment, that can persist for hours, suggesting vacuole maturation is uncoupled from membrane scission. Loss of PI(4,5)P2 is eventually triggered, a requisite step for lysosomal killing of the internalised cell. More broadly, PI(4,5)P2-positive vacuoles also form during T-cell engulfment by thymic nurse cells, dependent on ROCK activity, suggesting this distinctive compartment represents a shared feature of entosis-like cell engulfments.
]]></description>
<dc:creator>Durgan, J.</dc:creator>
<dc:creator>Sloan, K.</dc:creator>
<dc:creator>Domart, M.-C.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:creator>Florey, O.</dc:creator>
<dc:date>2024-10-22</dc:date>
<dc:identifier>doi:10.1101/2024.10.22.619613</dc:identifier>
<dc:title><![CDATA[A distinctive PI(4,5)P2 compartment forms during entosis and related engulfment processes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.23.619804v1?rss=1">
<title>
<![CDATA[
Imaging of live bacterial whole-cell biosensors illuminates spatial sialic acid availability within the inflamed mammalian gut 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.23.619804v1?rss=1"
</link>
<description><![CDATA[
Host mucin-derived sialic acids are key drivers of microbial colonisation, growth and pathogenicity within the mammalian gut. However, their study is hindered by complex spatiotemporal dynamics: Many gut metabolites, including sialic acids, are rapidly consumed, transformed or absorbed by the microbiota or host, meaning that conventional measurements of faeces or bulk digesta often fail to capture their true local availability. In contrast, engineered bacterial-whole cell biosensors provide an in situ read-out of metabolite exposure at the point and time of use, before these molecules are depleted. Here, we demonstrate increases in the bioavailability of sialic acid in the inflamed mouse gut using an engineered Escherichia coli biosensor that reports on sialic acid exposure via NanR-regulated transcriptional circuit. The biosensor robustly colonises the mouse gut and remains functional for at least six weeks. Through organ-scale imaging at single bacterial resolution we observe strong correlations between disease status and biosensor response in two inflammation models. Profiling along the length of the gut uncovers regional variations between the maximal sialic acid sensing and the peak inflammatory response within host tissues in a murine colitis model. Simultaneous tracking of host and microbial markers of inflammation informs the therapeutic response to sialidase inhibition, which accelerates disease recovery. Together, these data illuminate the complex spatial dynamics involved in shared host-microbiome metabolism and demonstrate the broader power of using engineered bacterial biosensors to monitor in situ bioavailability of rapidly turned-over metabolites within the gut.
]]></description>
<dc:creator>Carreno, D.</dc:creator>
<dc:creator>Robinson, C. M.</dc:creator>
<dc:creator>Jackson, R.</dc:creator>
<dc:creator>Li, P.</dc:creator>
<dc:creator>Nunes, V.</dc:creator>
<dc:creator>Palma-Duran, S. A.</dc:creator>
<dc:creator>Nye, E.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Riglar, D. T.</dc:creator>
<dc:date>2024-10-23</dc:date>
<dc:identifier>doi:10.1101/2024.10.23.619804</dc:identifier>
<dc:title><![CDATA[Imaging of live bacterial whole-cell biosensors illuminates spatial sialic acid availability within the inflamed mammalian gut]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.22.619657v1?rss=1">
<title>
<![CDATA[
Extrachromosomal DNA driven oncogene spatial heterogeneity and evolution in glioblastoma 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.22.619657v1?rss=1"
</link>
<description><![CDATA[
Oncogene amplification on extrachromosomal DNA (ecDNA) is strongly associated with treatment resistance and shorter survival for patients with cancer, including patients with glioblastoma. The non-chromosomal inheritance of ecDNA during cell division is a major contributor to intratumoral genetic heterogeneity. At present, the spatial dynamics of ecDNA, and the impact on tumor evolutionary trajectories, are not well understood. Here, we investigate the spatial-temporal evolution of ecDNA and its clinical impact by analyzing tumor samples from 94 treatment-naive human IDH-wildtype glioblastoma patients. We developed a spatial-temporal computational model of ecDNA positive tumors ( SPECIES) that integrates whole-genome sequencing, multi-region DNA FISH, and nascent RNAscope, to provide unique insight into the spatial dynamics of ecDNA evolution. Random segregation in combination with positive selection of ecDNAs induce large, predictable spatial patterns of cell-to-cell ecDNA copy number variation that are highly dependent on the oncogene encoded on the circular DNA. EGFR ecDNAs often reach high mean copy number (mean of 50 copies per tumor cell), are under strong positive selection (mean selection coefficient, s > 2) and do not co-amplify other oncogenes on the same ecDNA particles. In contrast, PDGFRA ecDNAs have lower mean copy number (mean of 15 copies per cell), are under weaker positive selection and frequently co-amplify other oncogenes on the same ecDNA. Evolutionary modeling suggests that EGFR ecDNAs often accumulate prior to clonal expansion. EGFR structural variants, including vIII and c-terminal deletions are under strong positive selection, are found exclusively on ecDNA, and are intermixed with wild-type EGFR ecDNAs. Simulations show EGFRvIII ecDNA likely arises after ecDNA formation in a cell with high wild-type EGFR copy number (> 10) before the onset of the most recent clonal expansion. This remains true even in cases of co-selection and co-amplification of multiple oncogenic ecDNA species in a subset of patients. Overall, our results suggest a potential time window in which early ecDNA detection may provide an opportunity for more effective intervention.

HighlightsO_LIecDNA is the most common mechanism of focal oncogene amplification in IDHwt glioblastoma.
C_LIO_LIEGFR and its variants on ecDNA are particularly potent, likely arising early in tumor development, providing a strong oncogenic stimulus to drive tumorigenesis.
C_LIO_LIWild-type and variant EGFR ecDNA heteroplasmy (co-occurrence) is common with EGFRvIII or c-terminal deletions being derived from EGFR wild-type ecDNA prior to the most recent clonal expansion.
C_LIO_LITumors with ecDNA amplified EGFR versus PDGFRA exhibit different evolutionary trajectories.
C_LIO_LISPECIES model can infer spatial evolutionary dynamics of ecDNA in cancer.
C_LIO_LIA delay between ecDNA accumulation and subsequent oncogenic mutation may give a therapeutic window for early intervention.
C_LI
]]></description>
<dc:creator>Noorani, I.</dc:creator>
<dc:creator>Haughey, M.</dc:creator>
<dc:creator>Luebeck, J.</dc:creator>
<dc:creator>Rowan, A.</dc:creator>
<dc:creator>Grönroos, E.</dc:creator>
<dc:creator>Terenzi, F.</dc:creator>
<dc:creator>Wong, I. T.-L.</dc:creator>
<dc:creator>Kittel, J.</dc:creator>
<dc:creator>Bailey, C.</dc:creator>
<dc:creator>Weeden, C.</dc:creator>
<dc:creator>Bell, D.</dc:creator>
<dc:creator>Joo, E.</dc:creator>
<dc:creator>Barbe, V.</dc:creator>
<dc:creator>Jones, M. G.</dc:creator>
<dc:creator>Nye, E.</dc:creator>
<dc:creator>Green, M.</dc:creator>
<dc:creator>Meader, L.</dc:creator>
<dc:creator>Norton, E. J.</dc:creator>
<dc:creator>Fabian, M.</dc:creator>
<dc:creator>Kanu, N.</dc:creator>
<dc:creator>Jamal-Hanjani, M.</dc:creator>
<dc:creator>Santarius, T.</dc:creator>
<dc:creator>Nicoll, J.</dc:creator>
<dc:creator>Boche, D.</dc:creator>
<dc:creator>Chang, H. Y.</dc:creator>
<dc:creator>Bafna, V.</dc:creator>
<dc:creator>Huang, W.</dc:creator>
<dc:creator>Mischel, P. S.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Werner, B.</dc:creator>
<dc:date>2024-10-25</dc:date>
<dc:identifier>doi:10.1101/2024.10.22.619657</dc:identifier>
<dc:title><![CDATA[Extrachromosomal DNA driven oncogene spatial heterogeneity and evolution in glioblastoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.29.620545v1?rss=1">
<title>
<![CDATA[
Sequential transcriptional programs underpin activation of quiescent hippocampal stem cells. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.29.620545v1?rss=1"
</link>
<description><![CDATA[
Postnatal neural stem cells are primarily quiescent, which is a cellular state that exists as a continuum from deep to shallow quiescence. The molecular changes that occur along this continuum are beginning to be understood but the transcription factor network governing these changes has not been defined. We show that these transitions are regulated by sequential transcription factor programs. Single-cell transcriptomic analyses of mice with loss- or gain-of-function of the essential activation factor Ascl1, reveal that Ascl1 promotes the activation of hippocampal neural stem cells by driving these cells out of deep quiescence, despite its low protein expression. Subsequently, during the transition from deep to shallow quiescence, Ascl1 induces the expression of Mycn, which drives progression through shallow states of quiescence towards an active state. Together, these results define the required sequence of transcription factors during hippocampal neural stem cell activation.
]]></description>
<dc:creator>Rigo, P.</dc:creator>
<dc:creator>Ahmed-de-Prado, S.</dc:creator>
<dc:creator>Johnston, R. L.</dc:creator>
<dc:creator>Choudhury, C.</dc:creator>
<dc:creator>Guillemot, F.</dc:creator>
<dc:creator>Harris, L.</dc:creator>
<dc:date>2024-10-31</dc:date>
<dc:identifier>doi:10.1101/2024.10.29.620545</dc:identifier>
<dc:title><![CDATA[Sequential transcriptional programs underpin activation of quiescent hippocampal stem cells.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.30.621121v1?rss=1">
<title>
<![CDATA[
Replisome passage through the cohesin ring 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.30.621121v1?rss=1"
</link>
<description><![CDATA[
Following eukaryotic genome replication, the two newly synthesised sister chromatids remain paired by the ring-shaped cohesin complex, enabling their faithful segregation to daughter cells during cell divisions1. Cohesin topologically embraces DNA already before DNA replication2-5, and replisome passage through the cohesin ring is thought of as a fail-safe mechanism ensuring that cohesin entraps both replication products6,7. Whether replisomes indeed pass through cohesin rings remains unknown. Here, we use biochemical reconstitution4,8,9 and single molecule fluorescence microscopy to directly visualise replisome-cohesin encounters. We find that the translocating eukaryotic replicative Cdc45-Mcm2-7-GINS (CMG) helicase is frequently an obstacle to cohesin, but that the likelihood with which CMG passes cohesin increases in the presence of replisome components with known sister chromatid cohesion functions7,10-14, or by preventing cohesin from freely sliding along the template DNA. Cohesin retains topological DNA entrapment during CMG passage, suggesting that the helicase passes through the ring. The passage frequency increases further when fully reconstituted replisomes encounter cohesin rings, resulting in successful establishment of cohesion between the two replication products. Our findings demonstrate the existence of a simple mechanism that links genome replication with chromosome segregation, replisome passage through cohesin rings.
]]></description>
<dc:creator>Glaser, S.</dc:creator>
<dc:creator>Molodtsov, M. I.</dc:creator>
<dc:creator>Diffley, J. F.</dc:creator>
<dc:creator>Uhlmann, F.</dc:creator>
<dc:date>2024-11-01</dc:date>
<dc:identifier>doi:10.1101/2024.10.30.621121</dc:identifier>
<dc:title><![CDATA[Replisome passage through the cohesin ring]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.01.621508v1?rss=1">
<title>
<![CDATA[
DNA replication fork stabilization and restart mechanisms revealed by biochemical reconstitution 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.01.621508v1?rss=1"
</link>
<description><![CDATA[
Understanding how DNA replication forks stall and restart and how the DNA damage checkpoint prevents irreversible fork collapse in molecular detail are crucial for understanding how cells maintain stable genomes and how they prevent the genetic instability that drives cancer. Here we describe the reconstitution of fork stalling and restart with purified budding yeast proteins. After nucleotide depletion, leading strand DNA synthesis quickly stops but CMG helicase continues to unwind and Okazaki fragments continue to initiate on the lagging strand. Incomplete Okazaki fragments sequester PCNA, RFC and DNA polymerases {delta} and {varepsilon} which prevents normal DNA synthesis restart and exposes nascent DNA to nuclease attack. The DNA damage checkpoint limits this sequestration by restraining fork progression, which protects stalled forks from collapse and ensures restart.
]]></description>
<dc:creator>Canal, B.</dc:creator>
<dc:creator>Bertolin, A. P.</dc:creator>
<dc:creator>Lee, G. C.</dc:creator>
<dc:creator>Minamino, M.</dc:creator>
<dc:creator>Diffley, J. F. X.</dc:creator>
<dc:date>2024-11-03</dc:date>
<dc:identifier>doi:10.1101/2024.11.01.621508</dc:identifier>
<dc:title><![CDATA[DNA replication fork stabilization and restart mechanisms revealed by biochemical reconstitution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.01.621514v1?rss=1">
<title>
<![CDATA[
The mechanism of checkpoint-dependent DNA replication fork stabilization in human cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.01.621514v1?rss=1"
</link>
<description><![CDATA[
The DNA damage checkpoint is crucial for maintaining genome stability after genotoxic stress; without it, excess DNA replication origins are activated, stalled DNA replication forks cannot restart normally, high levels of DNA damage and single-stranded DNA (ssDNA) accumulate and cells cannot complete S phase. Preventing excess origin firing suppresses all these effects. Here we show that when replication is not restrained by a functional checkpoint, excess DNA synthesis sequesters the processivity factor PCNA and its loader RFC, preventing normal fork restart. Nascent DNA ends unprotected by RFC/PCNA are attacked by the Helicase-Like Transcription Factor (HLTF), causing irreversible replication fork collapse and hyperaccumulation of single-stranded DNA. This explains how the checkpoint stabilizes stalled replication forks and has implications for how origin firing is normally coordinated with fork progression. Loss of HLTF suppresses fork collapse and cell lethality in checkpoint-deficient cells, which has implications for how resistance to anti-checkpoint therapies may arise.
]]></description>
<dc:creator>Bertolin, A. P.</dc:creator>
<dc:creator>Canal, B.</dc:creator>
<dc:creator>Yekezare, M.</dc:creator>
<dc:creator>Zeng, J.</dc:creator>
<dc:creator>Instrell, R.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>Diffley, J. F. X.</dc:creator>
<dc:date>2024-11-03</dc:date>
<dc:identifier>doi:10.1101/2024.11.01.621514</dc:identifier>
<dc:title><![CDATA[The mechanism of checkpoint-dependent DNA replication fork stabilization in human cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.04.621342v1?rss=1">
<title>
<![CDATA[
Functionally important binding site for a volatile anesthetic in a voltage-gated sodium channel identified by X-ray crystallography 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.04.621342v1?rss=1"
</link>
<description><![CDATA[
Voltage-gated sodium channels (VGSCs), key mediators of excitability and synaptic transmission, are established and functionally relevant targets for volatile anaesthetic (VA) action. Using the structurally homologous prokaryotic VGSCs NavMs and NaChBac as models, we present a structure-function analysis of VGSC-VA interactions. We report that multiple VAs compete for binding sites on NavMs, and that these direct interactions mediate functional effects of sevoflurane on NavMs that mirror those attributed to VA effects in eukaryotic VGSCs, including human isoforms. Using X-ray crystallography, we determined the first atomic-resolution structure of a VA bound to a VGSC, showing sevoflurane displacing lipids to bind in an intramembranous hydrophobic pocket of NavMs. A conserved tyrosine residue within this binding site is critical for channel gating, and its substitution with alanine abolishes sevoflurane binding and selectively eliminates the characteristic anaesthetic-induced hyperpolarising shift of steady-state inactivation that reduces neuronal excitability at physiological membrane potentials. Finally, we provide evidence supporting VA action at the conserved sites in human VGSC isoforms. These findings define the first VA binding site in a VGSC. A membrane-mediated access pathway to the binding site leads to negative modulation of channel function that reduces neuronal activity and excitatory synaptic transmission in general anaesthesia.
]]></description>
<dc:creator>Hollingworth, D.</dc:creator>
<dc:creator>Herold, K. F.</dc:creator>
<dc:creator>Kelly, G.</dc:creator>
<dc:creator>Mykhaylyk, V. B.</dc:creator>
<dc:creator>Xiang, J.</dc:creator>
<dc:creator>Wallace, B. A.</dc:creator>
<dc:creator>Hemmings, H. C.</dc:creator>
<dc:date>2024-11-04</dc:date>
<dc:identifier>doi:10.1101/2024.11.04.621342</dc:identifier>
<dc:title><![CDATA[Functionally important binding site for a volatile anesthetic in a voltage-gated sodium channel identified by X-ray crystallography]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.07.622475v1?rss=1">
<title>
<![CDATA[
Metabolic and neuroactivity imbalances in plasma from aniridia patients with PAX6 haploinsufficiency 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.07.622475v1?rss=1"
</link>
<description><![CDATA[
PAX6 is a transcription factor crucial for the development of the eye, pancreas, and brain. Heterozygous variants resulting in PAX6 haploinsufficiency are the main genetic cause of congenital aniridia, characterized by both anterior and posterior ocular defects and sight loss. The extra-ocular features of PAX6 haploinsufficiency are becoming more widely recognised, with systemic manifestations like obesity, diabetes, and neurological/behavioural disorders being reported. In this study, we uncovered the metabolomic profile of the blood plasma from 25 PAX6-related aniridia patients compared to gender and age-matched controls. We found significant disruptions in lipid and energy metabolism, increased oxidative stress and neurotransmitters imbalances, as well as alterations linked to the gut microbiome. This study identified novel metabolic changes associated with PAX6 haploinsufficiency, providing evidence for the systemic aetiology of congenital aniridia and emphasizing the need for multidisciplinary management and further exploration into ocular and systemic therapeutic approaches.
]]></description>
<dc:creator>Cunha, D. L.</dc:creator>
<dc:creator>Kit, V.</dc:creator>
<dc:creator>Skinner, J.</dc:creator>
<dc:creator>Welch, A. A.</dc:creator>
<dc:creator>Moosajee, M.</dc:creator>
<dc:date>2024-11-08</dc:date>
<dc:identifier>doi:10.1101/2024.11.07.622475</dc:identifier>
<dc:title><![CDATA[Metabolic and neuroactivity imbalances in plasma from aniridia patients with PAX6 haploinsufficiency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.11.622998v1?rss=1">
<title>
<![CDATA[
Generation of morphologically distinct astrocyte subtypes from human iPSCs reveals a link between glial shape, function and reactivity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.11.622998v1?rss=1"
</link>
<description><![CDATA[
Astrocyte morphology in vivo is heterogeneous across different subtypes and dynamically changes in response to various stimuli. However, several questions on the mechanistic links between shape and function remain unanswered. Here, we developed an efficient protocol to generate pure populations of morphologically distinct human astrocytes in vitro, which we used for a systematic analysis of shape-function relationships. We performed a structural, molecular, and functional characterization of these populations and highlighted how their distinct morphologies mirror distinct functional and transcriptional patterns at the population level. We were also able to both correlate gene expression profiles of these morphologically distinct astrocyte subtypes with in vivo astrocytes in the human brain, and to validate our findings with primary isolated murine astrocytes in vitro. Moreover, we show that the observed morphological differences are correlated with changes in key cytoskeletal proteins, which offers a potential link to the observed functional differences. Finally, we demonstrated that different morphological subtypes of astrocytes have distinct reactivity responses to a common stimulus. This study offers a glimpse into the shape-function dynamics of human astrocytes, highlighting potential mechanistic links between cytoskeletal usage and astrocyte function, while also providing tools and datasets that will be useful for further studies into human glial biology in health and disease.
]]></description>
<dc:creator>O'Toole, K.</dc:creator>
<dc:creator>Guetta, L.</dc:creator>
<dc:creator>Urbanaviciute, P.</dc:creator>
<dc:creator>Suklai, P.</dc:creator>
<dc:creator>Wright, C.</dc:creator>
<dc:creator>Fisher, T.</dc:creator>
<dc:creator>Liddelow, S.</dc:creator>
<dc:creator>Luisier, R.</dc:creator>
<dc:creator>Marzi, S. J.</dc:creator>
<dc:creator>Serio, A.</dc:creator>
<dc:date>2024-11-11</dc:date>
<dc:identifier>doi:10.1101/2024.11.11.622998</dc:identifier>
<dc:title><![CDATA[Generation of morphologically distinct astrocyte subtypes from human iPSCs reveals a link between glial shape, function and reactivity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.14.623407v1?rss=1">
<title>
<![CDATA[
From naïve pluripotency to human neural organoids through a three-dimensional morphogenetic continuum 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.14.623407v1?rss=1"
</link>
<description><![CDATA[
Human central nervous system (CNS) development involves complex transitions from pluripotency to regionalised neural tissues. The early phases of this process are inaccessible in humans but can potentially be modelled in vitro using brain organoids, including to study neurodevelopmental disorders. However, current methods are based on post-implantation-like human pluripotent stem cells (hPSCs), which exhibit a hypermethylated state and show epigenetic memory retention. Here we developed a 3D model of human CNS development, starting from naive human induced PSCs (hiPSCs), which exhibit a hypomethylated pre-implantation-like state of pluripotency and develop into 3D neuroepithelial cysts in a timely morphogenetic continuum. Upon treatment with appropriate signalling cues, naive-derived neuroepithelial cysts can be specified toward different axial identities. Extended culture of anterior-specified organoids results in forebrain-like structures containing both dorsal and ventral neural precursors as well as mature neurons, exhibiting appropriate cellular diversity and functional properties. We applied this system to model Fragile X Syndrome (FXS), an epigenetically regulated neurodevelopmental disorder. We found that FXS patient-derived naive hiPSCs, initially demethylated at the Fmr1 locus, gradually underwent remethylation during organoid development. In addition, Fmr1 silencing started much earlier than can be detected by pre-natal analysis, and is concomitant with the development of mosaicisms. Our approach provides a new platform for studying human CNS development, including early epigenetic events and regional patterning, demonstrating the potential of naive hiPSC-derived organoids for modelling neurodevelopmental disorders with complex epigenetic regulation.

Highlights- single naive hiPSCs differentiate into 3D neuroepithelial cysts in a timely morphogenetic continuum
- signalling cues at appropriate developmental transitions can direct naive hiPSC- derived organoids to different regional identities of the human CNS
- naive hiPSC-derived forebrain organoids display cellular complexity representing both dorsal and ventral identities
- forebrain organoids from Fragile X Syndrome patients recapitulate the genetic instability and epigenetic dysregulation of Fmr1 locus.
]]></description>
<dc:creator>Laterza, C.</dc:creator>
<dc:creator>Cesare, E.</dc:creator>
<dc:creator>Stuart, H. T.</dc:creator>
<dc:creator>D'Ercole, M.</dc:creator>
<dc:creator>Gesualdo, A.</dc:creator>
<dc:creator>La Barbera, M. G.</dc:creator>
<dc:creator>Brignani, S.</dc:creator>
<dc:creator>Ribecco, C.</dc:creator>
<dc:creator>Polli, R.</dc:creator>
<dc:creator>Frison, R.</dc:creator>
<dc:creator>Angiolillo, S.</dc:creator>
<dc:creator>Maseta, A.</dc:creator>
<dc:creator>Gagliano, O.</dc:creator>
<dc:creator>Cacchiarelli, D.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Tanaka, E. M.</dc:creator>
<dc:creator>Murgia, A.</dc:creator>
<dc:creator>Elvassore, N.</dc:creator>
<dc:date>2024-11-15</dc:date>
<dc:identifier>doi:10.1101/2024.11.14.623407</dc:identifier>
<dc:title><![CDATA[From naïve pluripotency to human neural organoids through a three-dimensional morphogenetic continuum]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.11.08.515652v1?rss=1">
<title>
<![CDATA[
The AMPK-TORC1 signalling axis regulates caffeine-mediated DNA damage checkpoint override and cell cycle effects in fission yeast 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.08.515652v1?rss=1"
</link>
<description><![CDATA[
Caffeine is a widely consumed neuroactive substance. It can modulate cell cycle progression, override DNA damage checkpoint signalling and increase chronological lifespan (CLS) in various model systems. Early studies suggested that caffeine inhibits the phosphatidylinositol 3-kinase-related kinase (PIKK) Rad3 to override DNA damage-induced cell cycle arrest in fission yeast. We have previously suggested that caffeine modulates cell cycle progression and lifespan by inhibiting the Target of Rapamycin Complex 1 (TORC1). Nevertheless, whether this inhibition is direct or not, has remained elusive. TORC1 controls metabolism and mitosis timing by integrating nutrients and environmental stress response (ESR) signalling. Nutritional or other stresses activate the Sty1-Ssp1-Ssp2 (AMP-activated protein kinase complex, AMPK) pathway, which inhibits TORC1 and accelerates mitosis through Sck2 inhibition. Additionally, activation of the ESR pathway can extend lifespan in fission yeast. Here, we demonstrate that caffeine activates Ssp1, Ssp2 and the AMPK{beta} regulatory subunit Amk2 to advance mitosis. Ssp2 is phosphorylated in an Ssp1-dependent manner following exposure to caffeine. Furthermore, Ssp1 and Amk2, are required for resistance to caffeine under conditions of prolonged genotoxic stress. The effects of caffeine on DNA damage sensitivity are uncoupled from mitosis in AMPK pathway mutants. We propose that caffeine interacts synergistically with other genotoxic agents to increase DNA damage sensitivity. Our findings suggest that caffeine accelerates mitotic division and is beneficial for CLS through AMPK. Direct pharmacological targeting of AMPK may serve towards health span and lifespan benefits beyond yeasts, given the highly conserved nature of this key regulatory cellular energy sensor.
]]></description>
<dc:creator>Alao, J.-P.</dc:creator>
<dc:creator>Rallis, C.</dc:creator>
<dc:date>2022-11-08</dc:date>
<dc:identifier>doi:10.1101/2022.11.08.515652</dc:identifier>
<dc:title><![CDATA[The AMPK-TORC1 signalling axis regulates caffeine-mediated DNA damage checkpoint override and cell cycle effects in fission yeast]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.18.624170v1?rss=1">
<title>
<![CDATA[
RNA editing is a molecular clock in unmodified human cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.18.624170v1?rss=1"
</link>
<description><![CDATA[
Despite major advances in spatial RNA sequencing, the ability to extract temporal information in RNA sequencing experiments is still limited. Here, we describe Transcriptome Timestamping (T2), a system which harnesses naturally occurring A-to-I editing of RNA transcripts in unmodified human cells to infer transcriptional history. T2 provides age estimates for individual RNA transcripts, and serves as an endogenous molecular recorder, differentiating between complex transcriptional programs. We show that T2 can identify transient and transitional transcriptional programs in primary differentiating monocytes that are not apparent from gene expression analysis alone, including a regulatory module in the monocyte-to-macrophage transition that, to our knowledge, has not yet been described in humans. Finally, we show that T2 can also be applied to single cell data, allowing us to identify transcriptional programs in heterogeneous populations, such as asynchronously dividing cells. T2 is a scalable approach to temporal transcriptomics that can be applied to track the activity of thousands of genes in unmodified, primary human cells and tissues, with no genetic engineering.
]]></description>
<dc:creator>Ghareeb, A. E.</dc:creator>
<dc:creator>Bayne, J.</dc:creator>
<dc:creator>Wagen, A. Z.</dc:creator>
<dc:creator>Abdelhafid, A. M.</dc:creator>
<dc:creator>Miller, D.</dc:creator>
<dc:creator>Cubitt, L.</dc:creator>
<dc:creator>Meran, L.</dc:creator>
<dc:creator>Hill, C.</dc:creator>
<dc:creator>Cribbs, A. P.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Gaffney, E.</dc:creator>
<dc:creator>Coles, M.</dc:creator>
<dc:creator>Young, G.</dc:creator>
<dc:creator>Rodriques, S. G.</dc:creator>
<dc:date>2024-11-19</dc:date>
<dc:identifier>doi:10.1101/2024.11.18.624170</dc:identifier>
<dc:title><![CDATA[RNA editing is a molecular clock in unmodified human cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.19.623756v1?rss=1">
<title>
<![CDATA[
Progressive chromosome shape changes during cell divisions, recapitulated by loop capture simulations 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.19.623756v1?rss=1"
</link>
<description><![CDATA[
Mitotic chromosomes give genome portions the required compaction and mechanical stability for faithful inheritance during cell divisions. Here, we record human chromosome dimensions from their appearance in prophase over successive times in a mitotic arrest. Chromosomes first appear long and uniformly thin. Then, individual chromosome arms become discernible, which continuously shorten and thicken - the longer a chromosome arm, the thicker it becomes. The observed chromosome arm length to width relationship can be described by a power law with progressively increasing exponent. In the search for a molecular explanation of this behavior, the popular loop extrusion model provides no obvious means by which longer arms become thicker. Instead, we find that simulations of an alternative loop capture model recapitulate key features of our observations, including the gradually developing arm length to width relationship. Our analyses portray chromosomes as out-of-equilibrium structures in the process of transitioning towards, but on biologically relevant time scales not typically reaching, steady state.
]]></description>
<dc:creator>Kakui, Y.</dc:creator>
<dc:creator>Kusano, Y.</dc:creator>
<dc:creator>Clarence, T.</dc:creator>
<dc:creator>Lopez, M.</dc:creator>
<dc:creator>Hirota, T.</dc:creator>
<dc:creator>Khatri, B. S.</dc:creator>
<dc:creator>Uhlmann, F.</dc:creator>
<dc:date>2024-11-20</dc:date>
<dc:identifier>doi:10.1101/2024.11.19.623756</dc:identifier>
<dc:title><![CDATA[Progressive chromosome shape changes during cell divisions, recapitulated by loop capture simulations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.20.624489v1?rss=1">
<title>
<![CDATA[
Ancestry-specific gene expression in peripheral monocytes mediates risk of neurodegenerative disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.20.624489v1?rss=1"
</link>
<description><![CDATA[
It is hypothesised that peripheral immune states responding to regional environmental triggers contribute to central neurodegeneration. Region-specific genetic selection pressures require this hypothesis to be assessed in an ancestry specific manner. Here we utilise genome-wide association studies and expression quantitative trait loci from African, East Asian and European ancestries to show that genes causing neurodegeneration are preferentially expressed in innate rather than adaptive immune cells, and that expression of these genes mediates the risk of neurodegenerative disease in monocytes in an ancestry-specific manner.
]]></description>
<dc:creator>Wagen, A. Z.</dc:creator>
<dc:creator>Reynolds, R. H.</dc:creator>
<dc:creator>Nee Foo, J.</dc:creator>
<dc:creator>Fairbrother-Browne, A.</dc:creator>
<dc:creator>Gustavsson, E. K.</dc:creator>
<dc:creator>Gagliano-Turin, S. A.</dc:creator>
<dc:creator>Wood, N. W.</dc:creator>
<dc:creator>Blauwendraat, C. K.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:date>2024-11-22</dc:date>
<dc:identifier>doi:10.1101/2024.11.20.624489</dc:identifier>
<dc:title><![CDATA[Ancestry-specific gene expression in peripheral monocytes mediates risk of neurodegenerative disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.22.624643v1?rss=1">
<title>
<![CDATA[
Targeted CRISPR Screens Reveal Genes Essential for Cryptosporidium Survival in the Host Intestine 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.22.624643v1?rss=1"
</link>
<description><![CDATA[
The Cryptosporidium parasite is one of the leading causes of diarrheal morbidity and mortality in children, and adolescent infections are associated with chronic malnutrition. There are no vaccines available for protection and only one drug approved for treatment that has limited eKicacy. A major barrier to developing new therapeutics is a lack of foundational knowledge of Cryptosporidium biology, including which parasite genes are essential for survival and virulence. Here, we iteratively improve the tools for genetically manipulating Cryptosporidium and develop a targeted CRISPR-based screening method to rapidly assess how the loss of individual parasite genes influence survival in vivo. Using this method we examine the parasites pyrimidine salvage pathway and a set of leading Cryptosporidium vaccine candidates. From this latter group we determined the parasite gene known as Cp23 to be essential for survival, which was confirmed through inducible knockout in vitro and in vivo. Parasites deficient in Cp23 were able to replicate within and emerge from infected epithelial cells, yet unable to initiate gliding motility which is essential for the reinfection of neighbouring cells. The targeted screening method presented here is highly versatile and will enable researchers to more rapidly expand the knowledge base for Cryptosporidium infection biology, paving the way for new therapeutics.
]]></description>
<dc:creator>Watson, L. C.</dc:creator>
<dc:creator>Sala, K. A.</dc:creator>
<dc:creator>Bernitz, N.</dc:creator>
<dc:creator>Baumgartel, L.</dc:creator>
<dc:creator>Pallett, M. A.</dc:creator>
<dc:creator>Marzook, N. B.</dc:creator>
<dc:creator>Straker, L. C.</dc:creator>
<dc:creator>Peng, D.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:creator>Sateriale, A.</dc:creator>
<dc:date>2024-11-22</dc:date>
<dc:identifier>doi:10.1101/2024.11.22.624643</dc:identifier>
<dc:title><![CDATA[Targeted CRISPR Screens Reveal Genes Essential for Cryptosporidium Survival in the Host Intestine]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.19.624330v1?rss=1">
<title>
<![CDATA[
The ALS- and FTD-associated proteins Annexin A11 and CHMP2B act sequentially in membrane repair 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.19.624330v1?rss=1"
</link>
<description><![CDATA[
Maintenance of plasma membrane and organellar integrity is essential for cell viability. Cells must recognise damaged membranes and orchestrate repair programmes to preserve compartmentalisation. A variety of cellular factors including ESCRTs, Annexins, stress granules, lipids and proteins allowing vesicle and organelle fusion with damaged membranes have been reported to contribute to membrane repair. However, whether these factors operate independently or together to repair membranes is unclear. Here, we expose temporal differences and interdependencies in the recruitment of ESCRT-III and Annexin proteins to sites of membrane damage. We show that while Annexins are recruited immediately to sites of damage, ESCRT-III assembles only after membrane sealing. We show that ESCRT-III acts to shed damaged membranes from the cell and that FTD-and ALS-associated mutations in CHMP2B and ANXA11 compromise the repair process. These data present an integrated  sealing and healing model of events allowing membrane repair and restoration of membrane integrity.

One-Sentence Summary: A rubric of sealing and healing for ESCRT-mediated membrane repair
]]></description>
<dc:creator>Heffner, C. M.</dc:creator>
<dc:creator>Starling, G. P.</dc:creator>
<dc:creator>Isaacs, A.</dc:creator>
<dc:creator>Carlton, J. G.</dc:creator>
<dc:date>2024-11-21</dc:date>
<dc:identifier>doi:10.1101/2024.11.19.624330</dc:identifier>
<dc:title><![CDATA[The ALS- and FTD-associated proteins Annexin A11 and CHMP2B act sequentially in membrane repair]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.25.625278v1?rss=1">
<title>
<![CDATA[
Integration of hunger and hormonal state gates infant-directed aggression 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.25.625278v1?rss=1"
</link>
<description><![CDATA[
Social behaviour is profoundly shaped by internal physiological states. While significant progress has been made in understanding how individual states such as hunger, stress, or arousal modulate behaviour, animals experience multiple states at any given time. The neural mechanisms that integrate such orthogonal states--and how this integration affects behaviour--remain poorly understood. Here we report how hunger and estrous state converge on neurons in the medial preoptic area (MPOA) to shape infant-directed behaviour. We find that hunger promotes pup-directed aggression in normally non-aggressive virgin female mice. This behavioural switch occurs through inhibition of MPOA neurons, driven by the release of neuropeptide Y (NPY) from Agouti-related peptide-expressing neurons in the arcuate nucleus (ArcAgRP neurons). The propensity for hunger-induced aggression is set by reproductive state, with MPOA neurons detecting changes in progesterone (P4) to estradiol (E2) ratio across the estrous cycle. Hunger and estrous state converge on HCN (hyperpolarization-activated cyclic nucleotide-gated) channels, which sets the baseline activity and excitability of MPOA neurons. Using micro-endoscopic imaging, we confirm these findings in vivo, revealing that MPOA neurons encode a state for pup-directed aggression. This work thus provides a mechanistic understanding of how multiple physiological states are integrated to flexibly control social behaviour.
]]></description>
<dc:creator>Cao, M.</dc:creator>
<dc:creator>Ammari, R.</dc:creator>
<dc:creator>Chen, M. X.</dc:creator>
<dc:creator>Wai, P.</dc:creator>
<dc:creator>Sahni, A.</dc:creator>
<dc:creator>Liang, S.</dc:creator>
<dc:creator>Legrave, N.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Strom, M.</dc:creator>
<dc:creator>Kohl, J.</dc:creator>
<dc:date>2024-11-25</dc:date>
<dc:identifier>doi:10.1101/2024.11.25.625278</dc:identifier>
<dc:title><![CDATA[Integration of hunger and hormonal state gates infant-directed aggression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.26.625424v1?rss=1">
<title>
<![CDATA[
Sex chromosomes shape the transcriptional landscape of the preimplantation mouseembryo 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.26.625424v1?rss=1"
</link>
<description><![CDATA[
Sex chromosomes are emerging as key regulators of adult health and disease in males (XY) and females (XX), but their impact on embryo development is poorly understood. Using single-cell RNA sequencing (scRNA-seq) on wild type and aneuploid mouse embryos, we show that sex chromosomes significantly shape the preimplantation embryo transcriptional landscape. A hierarchy of effects are identified, distinctly mediated by the Y chromosome, the dosage of X chromosomes, X-chromosome imprinting, and by Xist, the non-coding RNA that initiates X-inactivation. The sex chromosomes have strong trans effects on autosomal gene expression throughout preimplantation development. The Y chromosome has an unexpectedly pronounced impact on the trophectoderm, the precursor of the placenta, and this property is shared with genes expressed from the inactive X chromosome. The paternal and maternal X chromosomes differentially promote preimplantation growth, and we identify multiple novel candidate X-linked imprinted genes mediating this effect. Our findings show that sex chromosomes impact the embryo from the beginning of life, long before the appearance of overt sex differences.
]]></description>
<dc:creator>Snell, D. M.</dc:creator>
<dc:creator>Varsally, W.</dc:creator>
<dc:creator>Courtois, A.</dc:creator>
<dc:creator>Menchero, S.</dc:creator>
<dc:creator>Munusamy, P.</dc:creator>
<dc:creator>Rietdijk, R.</dc:creator>
<dc:creator>Ojarikre, O. A.</dc:creator>
<dc:creator>Strohbuecker, S.</dc:creator>
<dc:creator>Kaya, H.</dc:creator>
<dc:creator>Sangrithi, M. N.</dc:creator>
<dc:creator>Turner, J. M.</dc:creator>
<dc:date>2024-11-26</dc:date>
<dc:identifier>doi:10.1101/2024.11.26.625424</dc:identifier>
<dc:title><![CDATA[Sex chromosomes shape the transcriptional landscape of the preimplantation mouseembryo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.28.625863v1?rss=1">
<title>
<![CDATA[
Sensing of extracellular L-Proline availability by the integrated stress response determines the outcome of cell competition. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.28.625863v1?rss=1"
</link>
<description><![CDATA[
Cell competition is a quality control acting from development to the adult that eliminates cells that are less-fit than their neighbours. How winner cells induce the elimination of losers during this process is poorly understood. Here, we address this question by studying the onset of differentiation in mouse, where cell competition eliminates 35% of embryonic cells. These loser cells have mitochondrial dysfunction, and we find that this causes amino-acid deprivation and activation of the integrated stress response (ISR), a pathway essential for their survival. We show that L-Proline is a key amino-acid sensed by the ISR and that in a competitive environment, winner cells induce increased L-Proline uptake in loser cells. This causes ISR repression and their elimination. Our results imply that cell competition is acting as a nutrient sensor, eliminating dysfunctional cells when amino acids are plentiful but sparing them in nutrient poor environments.
]]></description>
<dc:creator>Krishnan, S.</dc:creator>
<dc:creator>Lima, A.</dc:creator>
<dc:creator>Low, Y. T.</dc:creator>
<dc:creator>Montero, S. P.</dc:creator>
<dc:creator>Tan, S.</dc:creator>
<dc:creator>Gregorio, A. D.</dc:creator>
<dc:creator>Barreto, A. P.</dc:creator>
<dc:creator>Bowling, S.</dc:creator>
<dc:creator>Vousden, K.</dc:creator>
<dc:creator>Rodriguez, T. A.</dc:creator>
<dc:date>2024-11-28</dc:date>
<dc:identifier>doi:10.1101/2024.11.28.625863</dc:identifier>
<dc:title><![CDATA[Sensing of extracellular L-Proline availability by the integrated stress response determines the outcome of cell competition.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.28.625692v1?rss=1">
<title>
<![CDATA[
Cross-tissue isoform switches by human transcription factors suggest a widespread regulatory mechanism 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.28.625692v1?rss=1"
</link>
<description><![CDATA[
Many human genes, including transcription factor (TF) genes, produce alternative coding isoforms. Although some alternative coding TF isoforms have been experimentally studied, a genome-wide characterisation of all such isoforms in normal human physiology is still lacking. Here, we integrate RNA sequencing and protein domain data to reveal the expression levels, tissue specificity and putative functional classes of alternative coding isoforms of human TF genes. We found that isoforms lacking a DNA-binding domain (DBD) are annotated for almost a third of known TF genes, comprise 17% of all annotated TF isoforms and likely reverse the regulatory effects of DBD-containing isoforms. Considering all coding TF isoforms, we uncovered 536 switches between the highest-expressed alternative isoforms produced by 360 TF genes across 43 adult tissues. We predicted functional consequences of these switches using a functional annotation of structural domains present in switching isoforms. Furthermore, we found 45 isoform switches between the brain and non-brain tissues and interpreted six of these switches in detail. Finally, we assembled a literature-based compendium of human TF genes with experimentally studied alternative isoforms. In total, our results suggest that the switches of alternative coding TF isoforms between adult tissues represent a widespread, yet understudied, functional mechanism.
]]></description>
<dc:creator>Sidorov, S.</dc:creator>
<dc:creator>Pal, K.</dc:creator>
<dc:creator>Lenhard, B.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:date>2024-11-29</dc:date>
<dc:identifier>doi:10.1101/2024.11.28.625692</dc:identifier>
<dc:title><![CDATA[Cross-tissue isoform switches by human transcription factors suggest a widespread regulatory mechanism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.12.04.626561v1?rss=1">
<title>
<![CDATA[
The essential host genome for Cryptosporidium intracellular survival exposes metabolic dependencies that can be leveraged for treatment 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.04.626561v1?rss=1"
</link>
<description><![CDATA[
Mapping how pathogens interact with their host cells can reveal unexpected pathogen and host cell biology, paving the way for new treatments. Cryptosporidium is an intracellular parasite of intestinal epithelial cells, and a leading cause of diarrheal death and disease in infants worldwide. Despite this, very little is known about the cell biology of infection of this eukaryotic pathogen. Here, we designed and implemented a unique microscopy-based arrayed CRISPR-Cas9 screen to interrogate the effects of the loss of every protein-coding human gene on a Cryptosporidium infection. As the experimental readout is image-based, we extracted multiple phenotypic features of infection, including parasite growth, progression of the parasite to its sexual life stage, and recruitment of host actin to  pedestals beneath the parasite vacuole. Using this dataset, we discovered a tipping point in the host cholesterol biosynthesis pathway that controls Cryptosporidium infection. Parasite growth can either be inhibited or promoted by the intermediary metabolite squalene. A build-up of squalene in epithelial cells creates a reducing environment, with more reduced host glutathione available for uptake by the parasite. Because Cryptosporidium has lost the ability to synthesise glutathione, this uptake from the host cell is required for growth and progression through its life cycle. We demonstrate that this dependency can be leveraged for treatment with the abandoned drug lapaquistat, an inhibitor of host squalene synthase that has efficacy against Cryptosporidium in vitro and in vivo.
]]></description>
<dc:creator>Marzook, N. B.</dc:creator>
<dc:creator>Song, O.-R.</dc:creator>
<dc:creator>Baumgärtel, L.</dc:creator>
<dc:creator>Bernitz, N.</dc:creator>
<dc:creator>Mkandawire, T. T.</dc:creator>
<dc:creator>Watson, L. C.</dc:creator>
<dc:creator>Nunes, V.</dc:creator>
<dc:creator>Warchal, S.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>Sateriale, A.</dc:creator>
<dc:date>2024-12-05</dc:date>
<dc:identifier>doi:10.1101/2024.12.04.626561</dc:identifier>
<dc:title><![CDATA[The essential host genome for Cryptosporidium intracellular survival exposes metabolic dependencies that can be leveraged for treatment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.28.625663v1?rss=1">
<title>
<![CDATA[
HIFalpha isoform specific activities drive cell-type specificity of VHL-associated oncogenesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.28.625663v1?rss=1"
</link>
<description><![CDATA[
Cancers arising from dysregulation of generally operative signaling pathways are often tissue specific, but the mechanisms underlying this paradox are poorly understood. Based on striking cell-type specificity, we postulated that these mechanisms must operate early in cancer development and set out to study them in a model of von Hippel Lindau (VHL) disease. Biallelic mutation of the VHL ubiquitin ligase leads to constitutive activation of hypoxia inducible factors HIF1A and HIF2A and is generally a truncal event in clear cell renal carcinoma. We used an oncogenic tagging strategy in which VHL-mutant cells are marked by tdTomato, enabling their observation, retrieval, and analysis early after VHL-inactivation. Our findings reveal markedly different consequences of HIF1A and HIF2A activation, but that both contribute to renal cell-type specific consequences of VHL-inactivation in the kidney. Early involvement of HIF2A in promoting proliferation within proximal tubular epithelium supports therapeutic targeting of HIF2A early in VHL disease.
]]></description>
<dc:creator>Lima, J. D. C. C.</dc:creator>
<dc:creator>Hooker, M.</dc:creator>
<dc:creator>Li, R.</dc:creator>
<dc:creator>Barros, A. B.</dc:creator>
<dc:creator>Masson, N.</dc:creator>
<dc:creator>Pugh, C. W.</dc:creator>
<dc:creator>Mole, D. R.</dc:creator>
<dc:creator>Adam, J.</dc:creator>
<dc:creator>Ratcliffe, P. J.</dc:creator>
<dc:creator>Kurlekar, S.</dc:creator>
<dc:date>2024-12-03</dc:date>
<dc:identifier>doi:10.1101/2024.11.28.625663</dc:identifier>
<dc:title><![CDATA[HIFalpha isoform specific activities drive cell-type specificity of VHL-associated oncogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.12.04.625416v1?rss=1">
<title>
<![CDATA[
Non canonical Sun1-Allan complex orchestrates nuclear envelope remodelling and Basal Body/MTOC Segregation during rapid mitosis in Plasmodium 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.04.625416v1?rss=1"
</link>
<description><![CDATA[
Mitosis in eukaryotes involves reorganization of the nuclear envelope (NE) and microtubule-organizing centres (MTOCs). During male gametogenesis in Plasmodium, the causative agent of malaria, mitosis is exceptionally rapid and highly divergent. Within 8 min, the haploid male gametocyte genome undergoes three replication cycles (1N to 8N), while maintaining an intact NE. Axonemes assemble in the cytoplasm and connect to a bipartite MTOC-containing nuclear pole (NP) and cytoplasmic basal body, producing eight flagellated gametes. The mechanisms coordinating NE remodelling, MTOC dynamics, and flagellum assembly remain poorly understood.

We identify the SUN1-ALLAN complex as a novel mediator of NE remodelling and bipartite MTOC coordination during Plasmodium male gametogenesis. SUN1, a conserved NE protein, localizes to dynamic loops and focal points at the nucleoplasmic face of the spindle poles. ALLAN, a divergent allantoicase, has a location like that of SUN1, and these proteins form a unique complex, detected by live-cell imaging, ultrastructural expansion microscopy, and interactomics. Deletion of either SUN1 or ALLAN genes disrupts nuclear MTOC organization, leading to basal body mis-segregation, defective spindle assembly, and impaired spindle microtubule-kinetochore attachment, but axoneme formation remains intact. Ultrastructural analysis revealed nuclear and cytoplasmic MTOC miscoordination, producing aberrant flagellated gametes lacking nuclear material. These defects block development in the mosquito and parasite transmission, highlighting the essential functions of this complex.
]]></description>
<dc:creator>Zeeshan, M.</dc:creator>
<dc:creator>Blatov, I.</dc:creator>
<dc:creator>Yanase, R.</dc:creator>
<dc:creator>Ferguson, D. J. P.</dc:creator>
<dc:creator>Pashley, S. L.</dc:creator>
<dc:creator>Chahine, Z.</dc:creator>
<dc:creator>Botte, Y. Y.</dc:creator>
<dc:creator>Mishra, A.</dc:creator>
<dc:creator>Marche, B.</dc:creator>
<dc:creator>Bhanvadia, S.</dc:creator>
<dc:creator>Hair, M.</dc:creator>
<dc:creator>Batra, S.</dc:creator>
<dc:creator>Markus, R.</dc:creator>
<dc:creator>Brady, D.</dc:creator>
<dc:creator>Bottrill, A.</dc:creator>
<dc:creator>vaughan, s.</dc:creator>
<dc:creator>Botte, C. Y.</dc:creator>
<dc:creator>Le Roch, K. G.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:creator>Tromer, E. C.</dc:creator>
<dc:creator>Tewari, R.</dc:creator>
<dc:date>2024-12-05</dc:date>
<dc:identifier>doi:10.1101/2024.12.04.625416</dc:identifier>
<dc:title><![CDATA[Non canonical Sun1-Allan complex orchestrates nuclear envelope remodelling and Basal Body/MTOC Segregation during rapid mitosis in Plasmodium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.12.02.626017v1?rss=1">
<title>
<![CDATA[
Epigenetic investigation of multifocal small intestinal neuroendocrine tumours reveals accelerated ageing of tumours and epigenetic alteration of metabolic genes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.02.626017v1?rss=1"
</link>
<description><![CDATA[
BackgroundSmall intestinal neuroendocrine tumours (SI-NETs) are the most common malignancy of the small intestine and around 50% of patients present in clinic with multifocal disease. Recent investigations into the genomic architecture of multifocal SI-NETs have found evidence that these synchronous primary tumours evolve independently of each other. They also have extremely low mutational burden and few known driver genes, suggesting that epigenetic dysregulation may be driving tumorigenesis. Very little is known about epigenetic gene regulation, metabolism and ageing in these tumours, and how these traits differ across multiple tumours within individual patients.

MethodsIn this study, we performed the first investigation of genome-wide DNA methylation in multifocal SI-NETs, assessing multiple primary tumours within each patient (n=79 primary tumours from 14 patients) alongside matched metastatic tumours (n=12) and normal intestinal epithelial tissue (n=9). We assessed multifocal SI-NET differential methylation using a novel method, comparing primary tumours with matched normal epithelial tissue and an enterochromaffin-enriched cell line to enrich for tumour-specific effects. This method reduced the identification of  false positive methylation differences driven by cell composition differences between tumour and normal epithelial tissue. We also assessed tumour ageing using epigenetic clocks and applied metabolic predictors in the dataset to assess methylation variation across key metabolic genes.

ResultsWe have identified 12,392 tumour-specific differentially methylated positions (Bonferroni corrected p<0.05) which were enriched for neural pathways. The expression levels of the genes associated with top sites were also found to be significantly altered in SI-NETs. Age acceleration was observed across SI-NETs and a variability in epigenetic  age of tumours within each patient, which we believe is reflecting the  order in which tumours have developed. This is supported by the correlation of age acceleration with somatic mutational count in the tumours. We have identified SI-NET associated alterations to the methylation patterns in key metabolic genes compared to matched normal tissue, which is more pronounced in metastatic tumours and tumours harbouring chromosome 18 loss of heterozygosity, indicating metabolic differences in these tumour subtypes.

ConclusionsWe have identified accelerated ageing and changes to regulation of metabolic genes, alongside an epigenetic signature of multifocal SI-NETs. These findings add to our understanding of multifocal SI-NET biology and their molecular differences which may be instrumental in the development of these elusive tumours.
]]></description>
<dc:creator>Webster, A. P.</dc:creator>
<dc:creator>Makinen, N.</dc:creator>
<dc:creator>Mensah, N.</dc:creator>
<dc:creator>Castignani, C.</dc:creator>
<dc:creator>Larose Cadieux, E.</dc:creator>
<dc:creator>Shivdasani, R.</dc:creator>
<dc:creator>Singh, P.</dc:creator>
<dc:creator>Vaikkinen, H.</dc:creator>
<dc:creator>Dhami, P.</dc:creator>
<dc:creator>Ecker, S.</dc:creator>
<dc:creator>Brown, M.</dc:creator>
<dc:creator>Rimmer, B.</dc:creator>
<dc:creator>Henderson, S.</dc:creator>
<dc:creator>Herrero, J.</dc:creator>
<dc:creator>Suderman, M.</dc:creator>
<dc:creator>Yousefi, P. D.</dc:creator>
<dc:creator>Beck, S.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>Nakakura, E.</dc:creator>
<dc:creator>Thirlwell, C.</dc:creator>
<dc:date>2024-12-05</dc:date>
<dc:identifier>doi:10.1101/2024.12.02.626017</dc:identifier>
<dc:title><![CDATA[Epigenetic investigation of multifocal small intestinal neuroendocrine tumours reveals accelerated ageing of tumours and epigenetic alteration of metabolic genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.12.09.627451v1?rss=1">
<title>
<![CDATA[
Divergent regulatory element programs steer sex-specific supporting cell differentiation along mouse gonadal development 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.09.627451v1?rss=1"
</link>
<description><![CDATA[
Gonadal sex determination relies on tipping a delicate balance involving the activation and repression of several transcription factors and signalling pathways. This is likely mediated by numerous non-coding regulatory elements that shape sex-specific transcriptomic programs. To explore the dynamics of these in detail, we performed paired time-series of transcriptomic and chromatin accessibility assays on pre-granulosa and Sertoli cells throughout their development in the embryo, making use of new and existing mouse reporter lines. Regulatory elements were associated with their putative target genes by linkage analysis, and this was complemented and verified experimentally using promoter capture Hi-C. We identified the transcription factor motifs enriched in these regulatory elements along with their occupancy, pinpointing LHX9/EMX2 as potentially critical regulators of ovarian development. Variations in the DNA sequence of these regulatory elements are likely to be responsible for many of the unexplained cases of individuals with Differences of Sex Development.

TeaserMultiomics analysis revealed the regulatory elements and transcription factors responsible for gonadal sex determination.
]]></description>
<dc:creator>Stevant, I.</dc:creator>
<dc:creator>Abberbock, E.</dc:creator>
<dc:creator>Ridnik, M.</dc:creator>
<dc:creator>Weiss, R.</dc:creator>
<dc:creator>Swisa, L.</dc:creator>
<dc:creator>Futtner, C.</dc:creator>
<dc:creator>Maatouk, D.</dc:creator>
<dc:creator>Lovell-Badge, R.</dc:creator>
<dc:creator>Malysheva, V.</dc:creator>
<dc:creator>Gonen, N.</dc:creator>
<dc:date>2024-12-12</dc:date>
<dc:identifier>doi:10.1101/2024.12.09.627451</dc:identifier>
<dc:title><![CDATA[Divergent regulatory element programs steer sex-specific supporting cell differentiation along mouse gonadal development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.12.16.628661v1?rss=1">
<title>
<![CDATA[
Modelling co-development between somites and neural tube with human Trunk-like Structures (hTLS) 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.16.628661v1?rss=1"
</link>
<description><![CDATA[
Human stem cell-based embryo models have opened new avenues of research in development by providing experimentally amenable in vitro systems. One of the features of embryo models is their multilineage differentiation, which allows the co-development of, and interactions between, tissues. Here, we utilise a human Trunk-like Structure (hTLS) model to explore trunk development. We show that hTLS have morphologically organised somites and a neural tube that form through self-organised, endogenous signalling including anteroposterior FGF, Wnt and Retinoic Acid (RA) gradients that modulate the fate of neuromesodermal progenitors. Comparison to an existing dataset from organogenesis-stage human embryos shows that hTLS cells approximate Carnegie Stage 13-14 (28-35 days post-fertilisation). The absence of a notochord leads to a dorsal identity, but exogenous exposure to smoothened agonist (promoting Sonic Hedgehog signalling) progressively ventralises both somites and neural tube in a dose-dependent manner. Furthermore, we identify endogenous signalling from the neural tube to the somites, which leads to medially localised ALDH1A2, and subsequent RA signalling from the somites to the neural tube, which leads to spontaneous neural progenitor patterning and PAX6 expression. Together, our data highlight the value of modularity in embryo models which we leverage to explore human trunk co-development.

SummaryUsing a 3D, stem cell-based model of human embryonic trunk development, we examine the interactions across somitic and neural tissues to better understand the dynamics of human trunk co-development.
]]></description>
<dc:creator>Makwana, K.</dc:creator>
<dc:creator>Tilley, L.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Thompson, J.</dc:creator>
<dc:creator>Baillie-Benson, P.</dc:creator>
<dc:creator>Rodriguez-Polo, I.</dc:creator>
<dc:creator>Moris, N.</dc:creator>
<dc:date>2024-12-17</dc:date>
<dc:identifier>doi:10.1101/2024.12.16.628661</dc:identifier>
<dc:title><![CDATA[Modelling co-development between somites and neural tube with human Trunk-like Structures (hTLS)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.12.24.630225v1?rss=1">
<title>
<![CDATA[
Mechanistic studies of autophagic cargo recruitment and membrane expansion through in vitro reconstitution 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.24.630225v1?rss=1"
</link>
<description><![CDATA[
Autophagy is a highly conserved catabolic pathway to remove deleterious cytosolic material to maintain cellular homeostasis and cell survival. Upon autophagy induction, a unique double-membraned structure, called a phagophore, forms and expands into a cup shape to engulf these cytosolic substrates. ATG8 proteins are covalently conjugated to autophagic membranes by lipidation of phosphatidylethanolamine (PE) and are thought to localise on both sides of the phagophore membrane. ATG8 conjugated on the inner membrane of the phagophore recruits autophagy cargo receptors, such as p62. To recapitulate events on the inner membrane, we used giant unilamellar vesicles (GUVs) as a model membrane and encapsulated proteins of interest inside GUVs, thus generating a membrane platform to which ATG8 proteins could be localised on the inner leaflet of the vesicles. We reconstituted WIPI2b-directed and cargo-directed ATG8 lipidation inside the GUVs and revealed distinct roles of WIPI2b and p62 in initiating the ATG conjugation cascade. Furthermore, we showed that p62 or p62 droplets were recruited to the inner membrane of the GUVs though interaction with membrane-bound ATG8s. Using a bead-based membrane expansion assay, we demonstrated a redistribution and local enrichment of membrane-bound ATG8s across the membrane upon interaction with p62 and p62 droplets. Our study provides novel model systems to investigate the interactions on the inner membrane of the phagophore and reveals fundamental molecular insights into phagophore membrane bending. This process is directed by ATG8-cargo interaction, during which cargo receptors concentrate ATG8 proteins on the inner surface of the phagophore membrane.
]]></description>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Litschel, T.</dc:creator>
<dc:creator>D'Antuono, R.</dc:creator>
<dc:creator>Davis, C.</dc:creator>
<dc:creator>Schreiber, A.</dc:creator>
<dc:creator>Tooze, S.</dc:creator>
<dc:date>2024-12-24</dc:date>
<dc:identifier>doi:10.1101/2024.12.24.630225</dc:identifier>
<dc:title><![CDATA[Mechanistic studies of autophagic cargo recruitment and membrane expansion through in vitro reconstitution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.12.23.630104v1?rss=1">
<title>
<![CDATA[
Structural and functional characterization of the KHNYN extended-diKH domain for mediating ZAP antiviral activity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.23.630104v1?rss=1"
</link>
<description><![CDATA[
Zinc finger antiviral protein (ZAP) binds CpG dinucleotides in viral RNA and targets them for decay. ZAP interacts with several cofactors to form the ZAP antiviral system, including KHNYN, a multidomain endoribonuclease required for ZAP-mediated RNA decay. However, it is unclear how the individual domains in KHNYN contribute to its activity. Here, we demonstrate that the KHNYN amino terminal extended-diKH (ex-diKH) domain is required for antiviral activity and present its crystal structure. The structure belongs to a rare group of KH-containing domains, characterized by a non-canonical arrangement between two type-1 KH modules, with an additional helical bundle. N4BP1 is a KHNYN paralog with an ex-diKH domain that functionally complements the KHNYN ex-diKH domain. Interestingly, the ex-diKH domain structure is present in N4BP1-like proteins in lancelets, which are basal chordates, indicating that it is evolutionarily ancient. While many KH domains demonstrate RNA binding activity, biolayer interferometry and electrophoretic mobility shift assays indicate that the KHNYN ex-diKH domain does not bind RNA. Furthermore, residues required for canonical KH domains to bind RNA are not required for KHNYN antiviral activity. By contrast, an inter-KH domain cleft in KHNYN is a potential protein-protein interaction site and mutations that eliminate arginine salt bridges at the edge of this cleft decrease KHNYN antiviral activity. This suggests that this domain could be a binding site for an unknown KHNYN cofactor.
]]></description>
<dc:creator>Youle, R. L.</dc:creator>
<dc:creator>Lista, M. J.</dc:creator>
<dc:creator>Bouton, C.</dc:creator>
<dc:creator>Kunzelmann, S.</dc:creator>
<dc:creator>Wilson, H.</dc:creator>
<dc:creator>Cottee, M. A.</dc:creator>
<dc:creator>Purkiss, A. G.</dc:creator>
<dc:creator>Morris, E. R.</dc:creator>
<dc:creator>Neil, S. J.</dc:creator>
<dc:creator>Taylor, I. A.</dc:creator>
<dc:creator>Swanson, C. M.</dc:creator>
<dc:date>2024-12-23</dc:date>
<dc:identifier>doi:10.1101/2024.12.23.630104</dc:identifier>
<dc:title><![CDATA[Structural and functional characterization of the KHNYN extended-diKH domain for mediating ZAP antiviral activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.12.11.627489v1?rss=1">
<title>
<![CDATA[
Dominant RDH12-retinitis pigmentosa impairs photoreceptor development and cone function in retinal organoids 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.11.627489v1?rss=1"
</link>
<description><![CDATA[
Retinal dehydrogenase 12 (RDH12) is a photoreceptor NADPH-dependent retinal reductase enzyme, converting all-trans-retinal to all-trans-retinol. Heterozygous variants in RDH12 cause a rare autosomal dominant (AD) retinitis pigmentosa. As no disease models exist, we generated human induced pluripotent stem cell derived retinal organoids (RO) from a RDH12-AD patient (with pathogenic c.759del p.(Phe254Leufs*24) variant), alongside a healthy control (WT). RDH12-AD RO exhibited correct localisation of RDH12 to the photoreceptor inner segments up to week 44; transmission electron microscopy at week 37 showed photoreceptors were less abundant and shorter in length compared to WT. Visual cone function, retinol biosynthesis and the vitamin A pathway were also highly disrupted at week 44. Our study is the first to describe a RDH12-AD disease model with pathology at later stages of photoreceptor differentiation, in keeping with the milder disease course seen in humans. It provides insights into the aetiology and possible targets for future therapeutic development.
]]></description>
<dc:creator>Mejecase, C.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Owen, N.</dc:creator>
<dc:creator>Soro-Barrio, P.</dc:creator>
<dc:creator>Cheloni, R.</dc:creator>
<dc:creator>Nair, N.</dc:creator>
<dc:creator>Sarkar, H.</dc:creator>
<dc:creator>Toualbi, L.</dc:creator>
<dc:creator>Moosajee, M.</dc:creator>
<dc:date>2024-12-15</dc:date>
<dc:identifier>doi:10.1101/2024.12.11.627489</dc:identifier>
<dc:title><![CDATA[Dominant RDH12-retinitis pigmentosa impairs photoreceptor development and cone function in retinal organoids]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.12.09.627552v1?rss=1">
<title>
<![CDATA[
Bounded optimality of time investments in rats, mice, and humans 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.09.627552v1?rss=1"
</link>
<description><![CDATA[
Time is our scarcest resource. Allocating time optimally presents a universal challenge for all organisms because the future benefits of time investments are uncertain. We developed a normative framework for assessing bounded optimality in time allocation, emphasizing the accuracy of future predictions, independent of subjective costs and benefits. In a common decision task across humans, rats, and mice, we varied uncertainty by titrating ambiguous sensory evidence and measured the time each subject was willing to invest post-decision. We observed that all species and subjects invested more time when they were more likely to be correct, which reflected a statistical confidence of uncertain evidence. Time allocation strategy approached the lower bound of optimality, indicating an accurate decision-by-decision assessment of confidence in the likelihood that waiting will pay off - independent of the subjective payoff values and time costs. We demonstrate that an elementary algorithm based on a drift-diffusion process algorithm can implement this optimal time investment strategy. These results illuminate the computational mechanisms governing rational time investment, showing that humans, rats, and mice can maximize payoffs via confidence-guided time allocation.

HighlightsO_LIComputational and behavioral framework to assess bounded optimality of investments.
C_LIO_LIHumans, rats, and mice invest more time to obtain more likely payoffs, in proportion to statistical confidence.
C_LIO_LITime investment was close to optimal model predictions, reflecting bounded optimality of investments under uncertainty.
C_LIO_LIBounded-optimal time investment may be an evolutionary ancient adaptive behavioral strategy.
C_LI
]]></description>
<dc:creator>Ott, T.</dc:creator>
<dc:creator>Bosc, M.</dc:creator>
<dc:creator>Sanders, J. I.</dc:creator>
<dc:creator>Masset, P.</dc:creator>
<dc:creator>Kepecs, A.</dc:creator>
<dc:date>2024-12-15</dc:date>
<dc:identifier>doi:10.1101/2024.12.09.627552</dc:identifier>
<dc:title><![CDATA[Bounded optimality of time investments in rats, mice, and humans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.12.09.627463v1?rss=1">
<title>
<![CDATA[
Cell volume regulates terminal differentiation of cultured human epidermal keratinocytes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.09.627463v1?rss=1"
</link>
<description><![CDATA[
Differentiation of cultured human epidermal stem cells is regulated by interactions with the underlying substrate. Whereas differentiation is typically stimulated when keratinocytes are prevented from spreading, we previously identified two micron-scale topographical substrates that regulate differentiation of spread cells. On one substrate (S1), individual cells interact with small circular topographies, and differentiation is stimulated; on the other (S2), cells interact with larger triangular topographies, and differentiation is inhibited. By scanning electron microscopy we visualised substrate interactions at higher resolution than previously and using live cell imaging we established that induction of the differentiation marker involucrin did not involve transient cell rounding on S1. Bulk gene expression profiling did not reveal any differences between cells on S1 and S2 prior to the selective upregulation of differentiation markers at 12h on S1 and cell stiffness was lower on both S1 and S2 than on flat substrates. Nevertheless, cells on S2 differed from cells on flat and S1 substrates because they exhibited reduced cell volume, prompting us to explore whether cell volume could regulate differentiation independent of culture substrate. Treatment with polyethylene glycol (PEG) reduced cell volume and inhibited differentiation regardless of whether keratinocytes were seeded on flat, S1 or S2 substrates, micropatterned islands or in suspension. Conversely, treatment with deionised water increased cell volume and stimulated differentiation of substrate adherent keratinocytes. On flat substrates treatment with the Ca2+ chelator 1,2-bis-(2-aminophenoxy)ethane-N,N,N,N-tetraacetic acid acetoxymethyl ester or an inhibitor of the water channel aquaporin 3 blocked induction of differentiaton by deionised water, whereas the gadolinium3+, a stretch-activated calcium channel blocker, did not. Our studies identify a new mechanism by which keratinocyte-niche interactions regulate initiation of differentiation.
]]></description>
<dc:creator>Watt, F. M.</dc:creator>
<dc:creator>Zijl, S.</dc:creator>
<dc:creator>Hiratsuka, T.</dc:creator>
<dc:creator>Mobasseri, A.</dc:creator>
<dc:creator>Ebrahimkutty, M.</dc:creator>
<dc:creator>Boermel, M.</dc:creator>
<dc:creator>Garcia-Manyes, S.</dc:creator>
<dc:date>2024-12-09</dc:date>
<dc:identifier>doi:10.1101/2024.12.09.627463</dc:identifier>
<dc:title><![CDATA[Cell volume regulates terminal differentiation of cultured human epidermal keratinocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.12.23.630099v1?rss=1">
<title>
<![CDATA[
Gene network architecture, mutation and selection collectively drive developmental pattern evolvability and predictability 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.23.630099v1?rss=1"
</link>
<description><![CDATA[
A hallmark of development is the generation of spatial patterns driven by morphogen gradients and Gene Regulatory Networks (GRNs). Although the mechanistic basis by which GRNs orchestrate cellular responses and tissue patterning during development is well understood, their evolutionary dynamics remain less clear. Still, utations in regulatory elements that govern GRN-driven patterning are a key mechanism for patterning evolution. In this study, we use the de novo evolution of a stripe phenotype as a model framework to investigate the evolutionary dynamics associated with the emergence of new spatial gene expression boundaries and the adjustment of existing boundaries. To probe general principles of GRN-driven pattern evolution we introduce a new high-throughput theoretical framework that rapidly produces a comprehensive dataset of evolutionary trajectories. We leverage this large dataset to investigate the types of mutations that drive different phenotypic shifts in spatial patterning. Our findings suggest that the order in which mutations in gene-gene interactions appear, and the specific combination of gene-gene interactions that mutate together determine the evolvability of novel spatial gene expression patterns. We interpret our results in the context of epistatic effects that naturally arise in networks of interconnected genes, and show how contingencies and constraints emerge in our system. Our results elucidate the interplay between mutation and gene network organization, revealing how historical contingencies arise and impact the evolvability of GRNs and the predictability of their evolutionary outcomes.
]]></description>
<dc:creator>Booth, H.</dc:creator>
<dc:creator>Hadjivasiliou, Z.</dc:creator>
<dc:date>2024-12-23</dc:date>
<dc:identifier>doi:10.1101/2024.12.23.630099</dc:identifier>
<dc:title><![CDATA[Gene network architecture, mutation and selection collectively drive developmental pattern evolvability and predictability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.12.23.630113v1?rss=1">
<title>
<![CDATA[
Type I interferons induced upon respiratory viral infection impair lung metastatic initiation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.23.630113v1?rss=1"
</link>
<description><![CDATA[
Invasive breast cancer accounts for 7% of all cancer-related deaths, with the lungs being a common site of metastases. At the same time, lower respiratory tract infections are a common cause of morbidity and mortality worldwide. Acute viral respiratory infections induce transitional changes in the lung; however, the impact of these changes on metastasis initiation and cancer progression remains unclear. Using primary murine MMTV-PyMT breast cancer cells in an experimental lung metastasis model, we show that changes induced by respiratory syncytial virus (RSV) infection impair tumor cell seeding and early establishment in the lung, resulting in lower number of metastatic nodules. Furthermore, we demonstrate that this reduction of metastases is due to alterations in the lung environment mediated by type I interferons (IFNs) that are produced in response to RSV infection. Consistent with that notion, intranasal administration of recombinant IFN- recapitulates the anti-tumor effect of RSV infection. Type I IFNs change the lung cellular composition and induce an Interferon Stimulated Gene (ISG) driven response, creating an alveolar environment that is less supportive of tumor cell growth. Indeed, epithelial cells from mice infected with RSV or intranasally exposed to IFN-, are less supportive of tumor cell growth ex vivo. Altogether, our results suggest that type I IFNs induced by infection with some respiratory viruses perturb the lungs and consequently interfere with the ability of tumor cells to successfully initiate metastatic colonization.

SignificanceWomen diagnosed with metastatic breast cancer have a low survival rate. The lungs are a common metastatic site and are constantly exposed to viral pathogens, such as coronavirus, RSV and influenza virus. Thus, breast cancer and respiratory virus infection are likely to co-occur, but their interplay remains unclear. We show that type I interferons (IFNs), induced upon viral infection impair metastatic cancer cell seeding of mouse lungs. This is potentially via an effect of IFNs on lung epithelial cells, which become less supportive of early tumor cell proliferation. These findings indicate that viral infections and type I IFNs can alter the lung environment and impair implantation of metastatic cells, which could be explored to improve future cancer treatments.
]]></description>
<dc:creator>Farias, A.</dc:creator>
<dc:creator>Bridgeman, V.</dc:creator>
<dc:creator>Rodrigues, F.</dc:creator>
<dc:creator>Owen, A.</dc:creator>
<dc:creator>Ruhland, S.</dc:creator>
<dc:creator>Ferreira, R.</dc:creator>
<dc:creator>Mack, M.</dc:creator>
<dc:creator>Malanchi, I.</dc:creator>
<dc:creator>Johansson, C.</dc:creator>
<dc:date>2024-12-23</dc:date>
<dc:identifier>doi:10.1101/2024.12.23.630113</dc:identifier>
<dc:title><![CDATA[Type I interferons induced upon respiratory viral infection impair lung metastatic initiation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.12.17.629001v1?rss=1">
<title>
<![CDATA[
Covalent inhibitors of the RAS binding domain of PI3Ka impair tumor growth driven by RAS and HER2 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.17.629001v1?rss=1"
</link>
<description><![CDATA[
Genetic disruption of the RAS binding domain (RBD) of PI 3-kinase (PI3K) prevents the growth of mutant RAS driven tumors in mice and does not impact PI3Ks role in insulin mediated control of glucose homeostasis. Selectively blocking the RAS-PI3K interaction may represent an attractive strategy for treating RAS-dependent cancers as it would avoid the toxicity associated with inhibitors of PI3K lipid kinase activity such as alpelisib. Here we report compounds that bind covalently to cysteine 242 in the RBD of PI3K p110 and block the ability of RAS to activate PI3K activity. These inhibitors have a profound impact on the growth of RAS mutant and also HER2 over-expressing tumors, particularly when combined with other inhibitors of the RAS/MAPK pathway, without causing hyperglycemia.
]]></description>
<dc:creator>Klebba, J. E.</dc:creator>
<dc:creator>Roy, N.</dc:creator>
<dc:creator>Bernard, S. M.</dc:creator>
<dc:creator>Grabow, S.</dc:creator>
<dc:creator>Hoffman, M. A.</dc:creator>
<dc:creator>Miao, H.</dc:creator>
<dc:creator>Tamiya, J.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Berry, C.</dc:creator>
<dc:creator>Esparza-Oros, A.</dc:creator>
<dc:creator>Lin, R.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Pariollaud, M.</dc:creator>
<dc:creator>Parker, H.</dc:creator>
<dc:creator>Mochalkin, I.</dc:creator>
<dc:creator>Rana, S.</dc:creator>
<dc:creator>Snead, A. N.</dc:creator>
<dc:creator>Walton, E. J.</dc:creator>
<dc:creator>Wyrick, T. E.</dc:creator>
<dc:creator>Aitichson, E.</dc:creator>
<dc:creator>Bedke, K.</dc:creator>
<dc:creator>Brannon, J. C.</dc:creator>
<dc:creator>Chick, J. M.</dc:creator>
<dc:creator>Hee, K.</dc:creator>
<dc:creator>Horning, B. D.</dc:creator>
<dc:creator>Ismail, M.</dc:creator>
<dc:creator>Lamb, K. N.</dc:creator>
<dc:creator>Lin, W.</dc:creator>
<dc:creator>Metzger, J.</dc:creator>
<dc:creator>Pastuszka, M. K.</dc:creator>
<dc:creator>Pollock, J.</dc:creator>
<dc:creator>Sigler, J. J.</dc:creator>
<dc:creator>Tomaschko, M.</dc:creator>
<dc:creator>Tran, E.</dc:creator>
<dc:creator>Kinsella, T. M.</dc:creator>
<dc:creator>Molina-Arcas, M.</dc:creator>
<dc:creator>Simon, G. M.</dc:creator>
<dc:creator>Weinstein, D. S.</dc:creator>
<dc:creator>Downward, J.</dc:creator>
<dc:creator>Patricelli, M. P.</dc:creator>
<dc:date>2024-12-20</dc:date>
<dc:identifier>doi:10.1101/2024.12.17.629001</dc:identifier>
<dc:title><![CDATA[Covalent inhibitors of the RAS binding domain of PI3Ka impair tumor growth driven by RAS and HER2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.01.03.631007v1?rss=1">
<title>
<![CDATA[
Transcriptomic analysis of repeat expansion-ataxias uncovers distinct non-neuronal cell type-specific signatures of disease across the human brain 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.01.03.631007v1?rss=1"
</link>
<description><![CDATA[
Hereditary ataxias are a heterogeneous group of neurogenetic conditions characterised by the clinical syndrome of progressive loss of coordination from neurodegeneration of the cerebellum. A commonality across the most prevalent ataxias is the underlying disease mechanism secondary to expansions of short tandem DNA repeats. There is currently an incomplete understanding of the pathogenic mechanisms of these repeat expansion disorders, a core feature of which revolves around RNA-dysregulation. In this study, we used both bulk and single nuclear RNA-sequencing to study post-mortem brain tissue of human donors with a range of repeat-expansion ataxias to reveal further mechanistic insights.

We compared post-mortem paired cerebellar and frontal cortex tissue bulk RNA-sequencing data from 23 ataxia patients and 22 sex-, age-matched controls from two brain banks (spinocerebellar ataxia (SCA)1, SCA2, SCA6, SCA7, SCA17, Friedreichs ataxia (FRDA), and 7 cases with unknown molecular diagnoses). We analysed bulk RNA-sequencing data for transcript usage, differential and cell-type-specific expression to transcriptomically profile these diseases. We also generated single nuclear RNA-sequencing data of the cerebellum from donors with SCA1, SCA2, SCA6 and FRDA to decipher changes in cell type proportions in the disease state.

Using this approach, we found that: (i) despite the commonalities in the genetics of ataxia, there were components of their transcriptional signatures which were distinct; (ii) there were extensive transcriptional changes evident not only in the cerebellum but also the frontal cortex in ataxia cases; (iii) activation of immune and inflammatory pathways, as well as involvement of non-neuronal cell types was a feature of all ataxias to a lesser or greater extent.

This study provides a novel resource to understand the mechanisms of disease in ataxia. Furthermore, taken together, these results highlight immune pathways and the role of non-neuronal cell types as early and potentially important therapeutic targets. These findings provide a map of transcriptomic changes in ataxia to further understanding of the underlying pathogenesis.
]]></description>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Hicks, A.</dc:creator>
<dc:creator>Brenton, J.</dc:creator>
<dc:creator>Grant-Peters, M.</dc:creator>
<dc:creator>Reynolds, R. H.</dc:creator>
<dc:creator>Gustavsson, E. K.</dc:creator>
<dc:creator>Rocca, C.</dc:creator>
<dc:creator>Rocamora-Perez, G.</dc:creator>
<dc:creator>Garza, R.</dc:creator>
<dc:creator>Garcia-Ruiz, S.</dc:creator>
<dc:creator>Dominik, N.</dc:creator>
<dc:creator>Anderson, C.</dc:creator>
<dc:creator>Curless, T.</dc:creator>
<dc:creator>Montgomery, K.</dc:creator>
<dc:creator>Macpherson, H.</dc:creator>
<dc:creator>Nethisinghe, S.</dc:creator>
<dc:creator>Gavriouchkina, D.</dc:creator>
<dc:creator>Blunskyte-Hendley, M.</dc:creator>
<dc:creator>Fairbrother-Browne, A.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Luo, H.</dc:creator>
<dc:creator>Efthymiou, S.</dc:creator>
<dc:creator>Murphy, D.</dc:creator>
<dc:creator>Hinton, F.</dc:creator>
<dc:creator>Botia, J. A.</dc:creator>
<dc:creator>Cortese, A.</dc:creator>
<dc:creator>Wood, N.</dc:creator>
<dc:creator>Giunti, P.</dc:creator>
<dc:creator>Hardy, J.</dc:creator>
<dc:creator>Jakobsson, J.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Tucci, A.</dc:creator>
<dc:creator>McLean, C.</dc:creator>
<dc:creator>Jaunmuktane, Z.</dc:creator>
<dc:creator>Houlden, H.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:date>2025-01-03</dc:date>
<dc:identifier>doi:10.1101/2025.01.03.631007</dc:identifier>
<dc:title><![CDATA[Transcriptomic analysis of repeat expansion-ataxias uncovers distinct non-neuronal cell type-specific signatures of disease across the human brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.01.10.632130v1?rss=1">
<title>
<![CDATA[
Timed chromatin invasion during mitosis governs prototype foamy virus integration site selection and infectivity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.01.10.632130v1?rss=1"
</link>
<description><![CDATA[
Selection of a suitable chromatin environment during retroviral integration is a tightly regulated and multilayered process that involves interplay between viral and host factors. However, whether intrinsic chromatin dynamics during mitosis modulate retroviral genome invasion is currently poorly described. Direct interaction between the spumaretrovirus prototype foamy virus (PFV) Gag protein and cellular chromatin has been described as a major determinant for integration site selection. A previous Gag chromatin-binding site (CBS)-nucleosome co-crystal structure revealed an interaction with the histone H2A-H2B acidic patch via a highly conserved arginine anchor residue. Yet, the molecular mechanisms regulating Gag-chromatin capture during PFV infection remain obscure. Here, we investigated the kinetics of Gag-chromatin interactions during mitosis and proviral integration of PFV-infected synchronized cells. Using Gag CBS variant viruses, we showed that alteration of Gag affinity for nucleosome binding induced untimely chromatin tethering during mitosis, decreased infectivity and redistributed viral integration sites to markers associated with late replication timing of host chromosomes. Mutant Gag proteins were moreover defective in their ability to displace the histone H4 tail from the nucleosome acidic patch of highly condensed mitotic chromatin. These data indicate that the mitotic chromatin landscape during Gag-nucleosome interactions hosts PFV integration site selection determinants and that spumaretroviruses evolved high-affinity chromatin binding to overcome early mitosis chromatin condensation for optimal viral DNA tethering, integration and infection.
]]></description>
<dc:creator>Lagadec, F.</dc:creator>
<dc:creator>Singh, P. K.</dc:creator>
<dc:creator>Calmels, C.</dc:creator>
<dc:creator>Lapaillerie, D.</dc:creator>
<dc:creator>Lindemann, D.</dc:creator>
<dc:creator>Parissi, V.</dc:creator>
<dc:creator>Cherepanov, P.</dc:creator>
<dc:creator>Engelman, A. N.</dc:creator>
<dc:creator>Lesbats, P.</dc:creator>
<dc:date>2025-01-11</dc:date>
<dc:identifier>doi:10.1101/2025.01.10.632130</dc:identifier>
<dc:title><![CDATA[Timed chromatin invasion during mitosis governs prototype foamy virus integration site selection and infectivity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.01.09.631995v1?rss=1">
<title>
<![CDATA[
Endogenous gene tagging with FnCas9 to track and sort neural lineages from 3D cortical organoids 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.01.09.631995v1?rss=1"
</link>
<description><![CDATA[
Neural lineage tracing, or molecular dissection of lineage-specific brain cell types, is used in many labs to learn how neurons grow and mature. However, these studies depend on the growth and characterization of pure cultures, which takes a long time because of biochemical or fluorescence-based isolation through cell surface markers that overlap. These lineage-specific cells, however, can be efficiently sorted using endogenously expressed, fluorescently labeled marker genes. The labeled cell lines can be used not only to differentiate and purify different types of neurons but also to study the long-term development of neural lineages in two- and three-dimensional development models. In this study, we used an orthogonal Cas protein to generate human embryonic stem cell (hESC) lines with genetically labeled fluorescent barcodes for discrete neural lineages. We use these lines to successfully demonstrate spatial and temporal tracing of DCX-positive neuroblasts and immature neuronal cells within 2D neural cultures and 3D cortical organoids derived from human embryonic stem cells. This allowed the purification of endogenously tagged live neural cells from heterogeneous cortical organoids across multiple stages of development.
]]></description>
<dc:creator>Kumar, M.</dc:creator>
<dc:creator>Torras, B. T.</dc:creator>
<dc:creator>Guillemot, F.</dc:creator>
<dc:creator>Chakraborty, D.</dc:creator>
<dc:date>2025-01-13</dc:date>
<dc:identifier>doi:10.1101/2025.01.09.631995</dc:identifier>
<dc:title><![CDATA[Endogenous gene tagging with FnCas9 to track and sort neural lineages from 3D cortical organoids]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.01.14.632683v1?rss=1">
<title>
<![CDATA[
Cell heterogeneity and fate bistability drive tissue patterning during intestinal regeneration 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.01.14.632683v1?rss=1"
</link>
<description><![CDATA[
Tissue regeneration relies on the ability of cells to undergo de novo patterning. While tissue patterning has been viewed as the transition from initially identical un-patterned cells to an arrangement of different cell types, recent evidence suggests that initial heterogeneities between cells modulate tissue-scale pattern formation. Yet, how such heterogeneities arise and, thereafter, regulate cell type emergence in a population of cells is poorly understood. Using in vivo and in vitro mouse regenerative systems, we identify a critical tissue density that is required to induce heterogeneous inactivation of the mechanosensor YAP1. Experimental and biophysical approaches demonstrate that YAP1 cell-to-cell heterogeneity pre-patterns the first cell fate decision, via both chromatin remodelling and a supracellular feedback between FOXA1 and Delta-Notch signalling. This feedback motif induces cell fate bistability endowing memory to the system and the maintenance of patterns during homeostasis. These findings reveal a generalisable framework in which transient cell-to-cell heterogeneity, regulated by tissue-scale properties, serves as a critical control parameter for the emergence of cell fate and stable patterning during regeneration.
]]></description>
<dc:creator>Schwayer, C.</dc:creator>
<dc:creator>Barbiero, S.</dc:creator>
<dc:creator>Brückner, D. B.</dc:creator>
<dc:creator>Baader, C.</dc:creator>
<dc:creator>Repina, N. A.</dc:creator>
<dc:creator>Diaz, O. E.</dc:creator>
<dc:creator>Meylan, L. C.</dc:creator>
<dc:creator>Kalck, V.</dc:creator>
<dc:creator>Suppinger, S.</dc:creator>
<dc:creator>Yang, Q.</dc:creator>
<dc:creator>Schnabl, J.</dc:creator>
<dc:creator>Kilik, U.</dc:creator>
<dc:creator>Camp, J. G.</dc:creator>
<dc:creator>Stockinger, B.</dc:creator>
<dc:creator>Bühler, M.</dc:creator>
<dc:creator>Stadler, M. B.</dc:creator>
<dc:creator>Hannezo, E.</dc:creator>
<dc:creator>Liberali, P.</dc:creator>
<dc:date>2025-01-15</dc:date>
<dc:identifier>doi:10.1101/2025.01.14.632683</dc:identifier>
<dc:title><![CDATA[Cell heterogeneity and fate bistability drive tissue patterning during intestinal regeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.01.14.632924v1?rss=1">
<title>
<![CDATA[
Unlocking the Secrets of NSP3: AlphaFold2-assisted Domain Determination in SARS-CoV-2 Protein 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.01.14.632924v1?rss=1"
</link>
<description><![CDATA[
Non-structural protein 3 (nsp3) is crucial for the SARS-CoV-2 infection cycle. It is the largest protein of the virus, consisting of roughly 2000 residues, and a major drug target. However, due to its size, disordered regions, and transmembrane domains, the atomic structure of the whole protein has not yet been established. Only 10 out of its 16 domains were individually determined in experiments.

Here, we demonstrate how structural bioinformatics, AI-based fold prediction, and traditional experiments complement each other and can shed light on the makeup of this important protein, both in SARS-CoV-2 and related viruses. Our method can be generalized for other multi-domain proteins, so we describe it in detail.

Our prediction-based approach reveals a previously undescribed folded domain, which we could confirm experimentally. Our research also suggests a potential function of the nidovirus-wide conserved domain Y1: This domain may be involved in the assembly of nsp3, nsp4, and nsp6 into the hexameric pore, which was discovered by electron tomography and exports RNA into the cytosol. The Y1-hexamer, however, could not be expressed and purified on its own. We also provide a revised domain segmentation and nomenclature of nsp3 domains based on a compilation of previous research and our own findings.
]]></description>
<dc:creator>Edich, M.</dc:creator>
<dc:creator>Briggs, D.</dc:creator>
<dc:creator>Gao, Y.</dc:creator>
<dc:creator>Thorn, A.</dc:creator>
<dc:date>2025-01-15</dc:date>
<dc:identifier>doi:10.1101/2025.01.14.632924</dc:identifier>
<dc:title><![CDATA[Unlocking the Secrets of NSP3: AlphaFold2-assisted Domain Determination in SARS-CoV-2 Protein]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.01.17.632799v1?rss=1">
<title>
<![CDATA[
Evolutionary trajectories of immune escape across cancers 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.01.17.632799v1?rss=1"
</link>
<description><![CDATA[
Immune escape is a critical hallmark of cancer progression and underlies resistance to multiple immunotherapies. However, it remains unclear when the genetic events associated with immune escape occur during cancer development. Here, we integrate functional genomics studies of immunomodulatory genes with a tumor evolution reconstruction approach to infer the evolution of immune escape across 38 cancer types from the Pan-Cancer Analysis of Whole Genomes dataset. Different cancers favor mutations in different immunomodulatory pathways. For example, the antigen presentation machinery is highly mutated in colorectal adenocarcinoma, lung squamous cell carcinoma, and chromophobe renal cell carcinoma, and the protein methylation pathway is highly mutated in bladder transitional cell carcinoma and lung adenocarcinoma. We also observe different timing patterns in multiple immunomodulatory pathways. For instance, mutations impacting genes involved in cellular amino acid metabolism were more likely to happen late in pancreatic adenocarcinoma. Mutations in the glucocorticoid receptor regulatory network pathway tended to occur early, while mutations in the TNF pathways were more likely to occur late in B-cell non-Hodgkin lymphoma. Mutations in the NOD1/2 signaling pathway and DNA binding transcription factor activity tended to happen late in breast adenocarcinoma and ovarian adenocarcinoma. Together, these results delineate the evolutionary trajectories of immune escape in different cancer types and highlight opportunities for improved immunotherapy of cancer.

SignificanceDespite its critical role in cancer progression, the evolution of immune escape is poorly understood. We integrate functional genomics and tumor evolution reconstruction and infer immune escape trajectories across cancer types. Our results have important implications for developing biomarkers for immunoprevention and treatment strategies for immune escape of cancer.
]]></description>
<dc:creator>Chen, W.</dc:creator>
<dc:creator>Baker, T.</dc:creator>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Ogilvie, H. A.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>Gu, S.</dc:creator>
<dc:date>2025-01-18</dc:date>
<dc:identifier>doi:10.1101/2025.01.17.632799</dc:identifier>
<dc:title><![CDATA[Evolutionary trajectories of immune escape across cancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.01.20.633776v1?rss=1">
<title>
<![CDATA[
RAS-PI3K Pathway in CAFs Shapes Physicochemical Properties of Tumor ECM and restrains Tumor progression 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.01.20.633776v1?rss=1"
</link>
<description><![CDATA[
Cancer-associated fibroblasts (CAFs) are key regulators of the tumor microenvironment, promoting tumor progression through extracellular matrix (ECM) remodeling and paracrine signaling, but the signaling pathways controlling CAF function remain incompletely defined. Here, we demonstrate that RAS-PI3K signaling plays a central role in CAF activation and ECM remodeling by promoting collagen crosslinking, fibronectin organization, and glycoprotein deposition at least partially through the activation of YAP. Disruption of RAS-PI3K interaction in CAFs leads to structurally and mechanically defective ECMs that impair tumor cell adhesion, migration, and proliferation. In vivo, fibroblast-specific deletion of RAS-PI3K reduces tumor burden in different models of KRAS-driven lung cancer, limits ECM deposition, and enhances the response to chemotherapy and RAS-targeted therapies in lung adenocarcinoma models. These findings position RAS-PI3K signaling as a critical regulator of CAF function and ECM remodeling, highlighting a drug repurposing therapeutic strategy to disrupt tumor- stroma interactions and improve treatment outcomes.
]]></description>
<dc:creator>Cuesta, C.</dc:creator>
<dc:creator>Alcon-Perez, M.</dc:creator>
<dc:creator>Zheng, J.</dc:creator>
<dc:creator>Procel, N.</dc:creator>
<dc:creator>Cota, R. R.</dc:creator>
<dc:creator>Fennema, D.</dc:creator>
<dc:creator>Rosell, A.</dc:creator>
<dc:creator>Meson, D. L.</dc:creator>
<dc:creator>Martinez-Castedo, B.</dc:creator>
<dc:creator>Arafat, Y.</dc:creator>
<dc:creator>Sanz-Fraile, H.</dc:creator>
<dc:creator>Rajeeve, V.</dc:creator>
<dc:creator>Alonso, D.</dc:creator>
<dc:creator>Hynds, R. E.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Alcaraz, J.</dc:creator>
<dc:creator>Cutillas, P.</dc:creator>
<dc:creator>Reyes-Aldasoro, C. C.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Castellano, E.</dc:creator>
<dc:date>2025-01-22</dc:date>
<dc:identifier>doi:10.1101/2025.01.20.633776</dc:identifier>
<dc:title><![CDATA[RAS-PI3K Pathway in CAFs Shapes Physicochemical Properties of Tumor ECM and restrains Tumor progression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.01.28.635015v1?rss=1">
<title>
<![CDATA[
Capturing Nematic Order on Tissue Surfaces of Arbitrary Geometry 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.01.28.635015v1?rss=1"
</link>
<description><![CDATA[
A leading paradigm for understanding the large-scale behavior of tissues is via generalizations of liquid crystal physics; much like liquid crystals, tissues combine fluid-like, viscoelastic behaviors with local orientational order, such as nematic symmetry. Whilst aspects of quantitative agreement have been achieved for flat monolayers, the most striking features of tissue morphogenesis -- such as symmetry breaking, folding and invagination -- concern surfaces with complex curved geometries. As yet, however, characterizing such complex behaviors in three dimensions has been frustrated due to the absence of proper image analysis methods; current state-of-the-art methods almost exclusively rely on two-dimensional (2D) intensity projections of multiple image planes, which superimpose data and lose geometric information that can be crucial. Here, we describe an analysis pipeline that properly captures the nematic order of tissue surfaces of arbitrary geometry, which we demonstrate in the context of in vitro multicellular aggregates, and in vivo zebrafish hearts. For the former, we correlate the number of topological defects with the aggregates surface area and verify theoretical predictions, whilst for the latter, we link biological properties to physical concepts (Laplace pressure) through spatio-temporal correlations of the heart geometry with fluorescence signals of intracellular proteins. Our analysis enables access to the  hidden third dimension of conventional image acquisition via stacked 2D planes and highlights how such characterizations can deliver meaningful physical insight.
]]></description>
<dc:creator>Eckert, J.</dc:creator>
<dc:creator>Andrews, T. G.</dc:creator>
<dc:creator>Pollard, J.</dc:creator>
<dc:creator>Priya, R.</dc:creator>
<dc:creator>Yap, A. S.</dc:creator>
<dc:creator>Morris, R. G.</dc:creator>
<dc:date>2025-01-29</dc:date>
<dc:identifier>doi:10.1101/2025.01.28.635015</dc:identifier>
<dc:title><![CDATA[Capturing Nematic Order on Tissue Surfaces of Arbitrary Geometry]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.01.31.635869v1?rss=1">
<title>
<![CDATA[
Arp2/3-mediated bidirectional actin assembly by SPIN90 dimers in metazoans 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.01.31.635869v1?rss=1"
</link>
<description><![CDATA[
Branched actin networks nucleated by the Arp2/3 complex play critical roles in various cellular processes, from cell migration to intracellular transport. However, when activated by WISH/DIP/SPIN90 family proteins, Arp2/3 nucleates linear actin filaments. Unexpectedly, we found that human SPIN90 is a dimer that can nucleate bidirectional actin filaments. To understand the basis for this, we determined a 3 [A] resolution structure of human SPIN90-Arp2/3 complex nucleating actin filaments. Our structure shows that SPIN90 dimerises via a 3-helix bundle and interacts with two Arp2/3 complexes. Each SPIN90 molecule binds both Arp2/3 complexes to promote their activation. Our analysis demonstrates that single filament nucleation by Arp2/3 is mechanistically more like branch formation than previously appreciated. The dimerisation domain in SPIN90 orthologues is conserved in metazoans, suggesting that this mode of bidirectional nucleation is a common strategy to generate anti-parallel actin filaments.
]]></description>
<dc:creator>Liu, T.</dc:creator>
<dc:creator>Cao, L.</dc:creator>
<dc:creator>Mladenov, M.</dc:creator>
<dc:creator>Romet-Lemonne, G.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:creator>Moores, C.</dc:creator>
<dc:date>2025-01-31</dc:date>
<dc:identifier>doi:10.1101/2025.01.31.635869</dc:identifier>
<dc:title><![CDATA[Arp2/3-mediated bidirectional actin assembly by SPIN90 dimers in metazoans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.05.636656v1?rss=1">
<title>
<![CDATA[
Non-Redundant Roles of Topoisomerase 2α and 2β in the Cytosolic Replication of Vaccinia Virus 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.05.636656v1?rss=1"
</link>
<description><![CDATA[
Vaccinia virus is a large enveloped DNA virus, which, like all poxviruses, replicates in the cytoplasm of infected cells. Vaccinia was historically thought to encode all the proteins required for its replication. However, more recent findings have shown that nuclear host proteins are redirected to the cytoplasm to facilitate viral replication. Among these, topoisomerase 2 (TOP2A) and 2{beta} (TOP2B), which mediate nuclear transcription, DNA replication, and chromosome segregation are the most abundant host proteins associated with nascent viral genomes. Here, we investigate the mechanisms driving TOP2A and TOP2B cytoplasmic translocation and their role in viral replication. We found that early viral protein synthesis induces the cytosolic relocalization of both isoforms, which are subsequently recruited to viral factories by an interaction of their C-terminal domains with the viral ligase, A50. TOP2A promotes replication by interacting with the vaccinia DNA replication machinery. In contrast, TOP2B suppresses replication by enhancing the formation of double-stranded RNA and antiviral granules, containing components of the tRNA splicing ligase complex. Our analysis provides new insights into host-pathogen interactions during poxvirus infection and the role of topoisomerase 2 outside of the nucleus.
]]></description>
<dc:creator>Dalla Rosa, I.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:date>2025-02-06</dc:date>
<dc:identifier>doi:10.1101/2025.02.05.636656</dc:identifier>
<dc:title><![CDATA[Non-Redundant Roles of Topoisomerase 2α and 2β in the Cytosolic Replication of Vaccinia Virus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.07.637038v1?rss=1">
<title>
<![CDATA[
Insect wings arose with a genetic circuit that extends the useful range of a BMP morphogen 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.07.637038v1?rss=1"
</link>
<description><![CDATA[
Morphogens are produced by a subset of cells to trigger a signalling gradient that provides positional information to surrounding tissues. At increasing distances from the source, the dwindling number of morphogen molecules is expected to constrain the useful range of morphogen gradients. We have identified a genetic circuit that counteracts this limitation in developing wings of Drosophila by boosting BMP signalling at the distal end of the gradient without amplifying the signal near the source. This circuit involves Brinker, a transcription factor that represses BMP target genes while itself being repressed by BMP signalling. We suggest that temporal averaging inherent to the production of the inverse Brk gradient contributes to the enhancement of the positional information far from the Dpp source. Despite being a core component of BMP signalling in flies, Brinker is exclusively found in insects, likely in all insect species. Genomic analysis across a wide range of insects and gene expression analysis in limb primordia of the apterygote Thermobia domestica suggests that Brinker is an insect-specific innovation that was subsequently wired into the BMP signalling network in pterygotes, perhaps to enable wing development.
]]></description>
<dc:creator>Huang, A.</dc:creator>
<dc:creator>Cocconi, L.</dc:creator>
<dc:creator>Nicholls-Mindlin, B.</dc:creator>
<dc:creator>Alexandre, C.</dc:creator>
<dc:creator>Salbreux, G.</dc:creator>
<dc:creator>Vincent, J.-P.</dc:creator>
<dc:date>2025-02-08</dc:date>
<dc:identifier>doi:10.1101/2025.02.07.637038</dc:identifier>
<dc:title><![CDATA[Insect wings arose with a genetic circuit that extends the useful range of a BMP morphogen]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.10.633538v1?rss=1">
<title>
<![CDATA[
Self-supervised image restoration in coherent X-rayneuronal microscopy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.10.633538v1?rss=1"
</link>
<description><![CDATA[
Coherent X-ray microscopy is emerging as a transformative technology for neuronal imaging, with the potential to offer a scalable solution for reconstruction of neural circuits in millimeter sized tissue volumes. Specifically, X-ray holographic nanoto-mography (XNH) brings together outstanding capabilities in terms of contrast, spatial resolution and data acquisition speed. While recent XNH developments already enabled generating valuable datasets for neuro-sciences, a major challenge for reconstruction of neural circuits remained overcoming resolving power limits to distinguish smaller neurites and synapses in the reconstructed volumes. Here we present a self-supervised image restoration approach that simultaneously improves spatial resolution, contrast, and data acquisition speed. This enables revealing synapses with XNH, marking a major milestone in the quest for generating connectomes of full mammalian brains. We demonstrate that this method is effective for various types of neuronal tissues and acquisition schemes. We propose a scalable implementation compatible with multi-terabyte image volumes. Altogether, this work brings large-scale X-ray nanotomography to a new precision level.
]]></description>
<dc:creator>Laugros, A.</dc:creator>
<dc:creator>Cloetens, P.</dc:creator>
<dc:creator>Bosch, C.</dc:creator>
<dc:creator>Schoonhoven, R.</dc:creator>
<dc:creator>Pavlovic, L.</dc:creator>
<dc:creator>Kuan, A. T.</dc:creator>
<dc:creator>Livingstone, J.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Hendriksen, A.</dc:creator>
<dc:creator>Holler, M.</dc:creator>
<dc:creator>Wanner, A. A.</dc:creator>
<dc:creator>Azevedo, A.</dc:creator>
<dc:creator>Batenburg, K. J.</dc:creator>
<dc:creator>Tuthill, J. C.</dc:creator>
<dc:creator>Lee, W.-C. A.</dc:creator>
<dc:creator>Schaefer, A. T.</dc:creator>
<dc:creator>Vigano, N.</dc:creator>
<dc:creator>Pacureanu, A.</dc:creator>
<dc:date>2025-02-10</dc:date>
<dc:identifier>doi:10.1101/2025.02.10.633538</dc:identifier>
<dc:title><![CDATA[Self-supervised image restoration in coherent X-rayneuronal microscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.10.637409v1?rss=1">
<title>
<![CDATA[
Enhancing Reproducibility Through Bioimage Analysis: The Significance of Effect Sizes and Controls 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.10.637409v1?rss=1"
</link>
<description><![CDATA[
Bioimage analysis is a powerful tool for investigating complex biological processes, but its robustness depends on technical precision and rigorous experimental design. In particular, the use of appropriate controls and experimental repetition is critical for drawing meaningful conclusions. However, both are often used inadequately or overlooked, with "statistical significance" often prioritised, frequently obtained through the misuse or misinterpretation of statistical tests. In this study, we reanalyse publicly available image datasets to highlight the crucial role of robust experimental design in interpreting results. Our findings underscore the importance of focusing on effect sizes and biological relevance over arbitrary statistical thresholds. We also discuss the diminishing returns of increased data collection once statistical stability has been achieved. By refining control usage and emphasising effect sizes, this work aims to enhance the reproducibility and robustness of research findings. We provide open-access code to allow researchers to engage with the dataset, promoting better practices in experimental design and data interpretation.
]]></description>
<dc:creator>Barry, D. J.</dc:creator>
<dc:creator>Marcotti, S.</dc:creator>
<dc:creator>Gerontogianni, L.</dc:creator>
<dc:creator>Kelly, G.</dc:creator>
<dc:date>2025-02-11</dc:date>
<dc:identifier>doi:10.1101/2025.02.10.637409</dc:identifier>
<dc:title><![CDATA[Enhancing Reproducibility Through Bioimage Analysis: The Significance of Effect Sizes and Controls]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.06.636691v1?rss=1">
<title>
<![CDATA[
Generation of C9orf72h370 mice, an intron 1 humanised C9orf72 repeat-expansion knock-in model 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.06.636691v1?rss=1"
</link>
<description><![CDATA[
An autosomal dominant GGGGCC repeat expansion in intron 1 of the C9orf72 gene is the most common genetic cause of both amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Here, we set out to engineer a gene targeted mouse model harbouring a pathogenic length humanised C9orf72 repeat expansion allele, in order to model pathological mechanisms in a physiological context. In human disease, pathogenic repeats typically range from the hundreds to thousands of units in length, representing a considerable challenge for cellular and in vivo model generation given the instability of GC rich and repetitive DNA sequences during molecular cloning. To overcome this challenge, we developed new methodology to synthetically and iteratively build pure GGGGCC repeats within a linear vector system, which we then seamlessly and scarlessly embedded within the native human genomic sequence. This created a gene targeting DNA vector for homologous recombination of the human sequence in mouse embryonic stem cells. We used this novel targeting vector to generate a new gene targeted mouse allele, C9orf72h370, that for the first time has mouse C9orf72 intron 1 scarlessly replaced with human intron 1 including a pure (GGGGCC)370 hexanucleotide repeat expansion. We confirm that the mouse model expresses human intron 1-derived RNA and produces dipeptide repeat proteins derived from the GGGGCC repeat expansion. We now provide this model as a new freely available resource for the field. In addition, we demonstrate the utility of our cloning method for engineering diverse repeat expansion sequences for modelling other disorders, such as Fragile X Syndrome.
]]></description>
<dc:creator>Nair, R. R.</dc:creator>
<dc:creator>Carcole, M.</dc:creator>
<dc:creator>Thompson, D.</dc:creator>
<dc:creator>Tibbit, C.</dc:creator>
<dc:creator>McLeod, R.</dc:creator>
<dc:creator>Cammack, A.</dc:creator>
<dc:creator>Jakubcova, T.</dc:creator>
<dc:creator>Biggs, D.</dc:creator>
<dc:creator>Wyles, M.</dc:creator>
<dc:creator>Parker, M.</dc:creator>
<dc:creator>Caulder, A.</dc:creator>
<dc:creator>Teboul, L.</dc:creator>
<dc:creator>Fisher-Ward, C. L.</dc:creator>
<dc:creator>Awan, A. R.</dc:creator>
<dc:creator>Flower, M.</dc:creator>
<dc:creator>Davies, B.</dc:creator>
<dc:creator>Isaacs, A. M.</dc:creator>
<dc:creator>Fisher, E. M.</dc:creator>
<dc:creator>Cunningham, T. J.</dc:creator>
<dc:date>2025-02-06</dc:date>
<dc:identifier>doi:10.1101/2025.02.06.636691</dc:identifier>
<dc:title><![CDATA[Generation of C9orf72h370 mice, an intron 1 humanised C9orf72 repeat-expansion knock-in model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.11.637674v1?rss=1">
<title>
<![CDATA[
Functional specialisation of multisensory temporal integration in the mouse superior colliculus 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.11.637674v1?rss=1"
</link>
<description><![CDATA[
Our perception of the world depends on the brains ability to integrate information from multiple senses, with temporal disparities providing a critical cue for binding or segregating cross-modal signals1,2. The superior colliculus (SC) is a key site for integrating sensory modalities, but how cellular and network mechanisms in distinct anatomical regions within the SC contribute to multisensory integration remains poorly understood. Here, we recorded responses from over 5,000 neurons across the SCs anatomical axes of awake mice during presentations of spatially coincident audiovisual stimuli with varying temporal asynchronies. Our findings revealed that multisensory neurons reliably encoded audiovisual delays and exhibited nonlinear summation of auditory and visual inputs, with nonlinearities being more pronounced when visual stimuli preceded auditory stimuli, consistent with the natural statistics of light and sound propagation. Nonlinear summation was crucial for population-level decoding accuracy and precision of AV delay representation. Moreover, enhanced population decoding of audiovisual delays in the posterior-medial SC, facilitated temporal discriminability in the peripheral visual field. Cross-correlation analysis indicated higher connectivity in the medial SC and functional specific recurrent connectivity, with visual, auditory, and multisensory neurons preferentially connecting to other neurons of the same functional subclass, and multisensory neurons receiving approximately 50 percent of the total local input from other multisensory neurons. Our results highlight the interplay between single-neuron computations, network connectivity, and population coding in the SC, where nonlinear integration, distributed representations and regional functional specialisations enables robust sensory binding and supports the accurate encoding of temporal multisensory information. Our study provides new insights into how the brain leverages both single-neuron and network-level mechanisms to represent sensory features by adapting to the statistics of the natural world.
]]></description>
<dc:creator>Bianchini, G.</dc:creator>
<dc:creator>Razafindrahaba, I.</dc:creator>
<dc:creator>Moglie, M. J.</dc:creator>
<dc:creator>Konstantinou, G.</dc:creator>
<dc:creator>Cano-Ferrer, X.</dc:creator>
<dc:creator>Imbert, A.</dc:creator>
<dc:creator>Iacaruso, F.</dc:creator>
<dc:date>2025-02-12</dc:date>
<dc:identifier>doi:10.1101/2025.02.11.637674</dc:identifier>
<dc:title><![CDATA[Functional specialisation of multisensory temporal integration in the mouse superior colliculus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.04.636413v1?rss=1">
<title>
<![CDATA[
Enhancing the efficiency of oncolytic vaccinia virus for ovarian cancer treatment 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.04.636413v1?rss=1"
</link>
<description><![CDATA[
Vaccinia virus, known for its clinical safety has a tropism for primary and metastatic tumours as well as ovarian tissue. Consequently, oncolytic approaches with recombinant vaccinia viruses have emerged as attractive agents against ovarian cancer. Unfortunately, oncolytic vaccinia monotherapies are yet to live up to their potential promise. Given this, there is a need to identify combination agents that improve the effectiveness of vaccinia in ovarian cancer treatment. We screened 9,000 compounds to identify drugs that enhance the ability of a recombinant vaccinia virus lacking VGF and F1 ({Delta}VF) to induce death of ID8 Trp53-/- murine ovarian cancer cells. We identified a class of tubulin polymerisation inhibitors including vinorelbine. The combination of vinorelbine and vaccinia induces ID8 Trp53-/- cell death via apoptosis. In a syngeneic mouse model of high grade serous ovarian carcinoma, {Delta}VF virus lacking the viral thymidine kinase (TK), armed with GM-CSF and expressing NeonGreen ({Delta}VFTK-NG-GM-CSF) is tumour specific. A combination of the {Delta}VFTK-NG-GM-CSF virus with vinorelbine prolongs mouse survival compared to the treatment of mice with either agent alone. Our study suggests vinorelbine is a promising agent to combine with oncolytic vaccinia virus approaches for the management of ovarian cancer.
]]></description>
<dc:creator>Drymiotou, S.</dc:creator>
<dc:creator>Queval, C. J.</dc:creator>
<dc:creator>Tyson, K. E.</dc:creator>
<dc:creator>Sheach, L. A.</dc:creator>
<dc:creator>Postigo, A.</dc:creator>
<dc:creator>Rosa, I. D.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>McNeish, I. A.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:date>2025-02-04</dc:date>
<dc:identifier>doi:10.1101/2025.02.04.636413</dc:identifier>
<dc:title><![CDATA[Enhancing the efficiency of oncolytic vaccinia virus for ovarian cancer treatment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.15.638424v1?rss=1">
<title>
<![CDATA[
Subunit specialisation in AAA+ proteins and substrate unfolding during transcription complex remodelling 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.15.638424v1?rss=1"
</link>
<description><![CDATA[
Bacterial RNA polymerase (RNAP) is a multi-subunit enzyme that copies DNA into RNA in a process known as transcription. Bacteria use {sigma} factors to recruit RNAP to promoter regions of genes that need to be transcribed, with 60% bacteria containing at least one specialized {sigma} factor {sigma}54. {sigma}54 recruits RNAP to promoters of genes associated with stress responses and forms a stable closed complex that does not spontaneously isomerize to the open state where promoter DNA is melted out and competent for transcription. The {sigma}54-mediated open complex formation requires specific AAA+ proteins (ATPases Associated with diverse cellular Activities) known as bacterial enhancer-binding proteins (bEBPs). We have now obtained structures of new intermediate states of bEBP-bound complexes during transcription initiation, which elucidate the mechanism of DNA melting driven by ATPase activity of bEBPs and suggest a mechanistic model that couples the ATP hydrolysis cycle within the bEBP hexamer with {sigma}54 unfolding. Our data reveal that bEBP forms a non-planar hexamer with the hydrolysis-ready subunit located at the furthest/highest point of the spiral hexamer relative to the RNAP. ATP hydrolysis induces conformational changes in bEBP that drives a vectoral transiting of the regulatory N-terminus of {sigma}54 into the bEBP hexamer central pore causing the partial unfolding of {sigma}54, while forming specific bEBP contacts with promoter DNA. Furthermore, our data suggest a mechanism of AAA+ protein that is distinct from the hand-over-hand mechanism proposed for many AAA+ proteins, highlighting the versatile mechanisms utilized by the large protein family.
]]></description>
<dc:creator>Gao, F.</dc:creator>
<dc:creator>Ye, F.</dc:creator>
<dc:creator>Buck, M.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:date>2025-02-16</dc:date>
<dc:identifier>doi:10.1101/2025.02.15.638424</dc:identifier>
<dc:title><![CDATA[Subunit specialisation in AAA+ proteins and substrate unfolding during transcription complex remodelling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.21.639482v1?rss=1">
<title>
<![CDATA[
Phosphatase specificity influences phosphorylation timing of CDK substrates during the cell cycle 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.21.639482v1?rss=1"
</link>
<description><![CDATA[
Cell cycle events are ordered by cyclin-dependent kinases (CDKs), which phosphorylate hundreds of substrates. Multiple phosphatases oppose CDK substrate phosphorylation, yet a systematic understanding of how these phosphatases collectively influence phosphorylation timing is lacking. Here, we show that phosphatases influence the timing of CDK substrate phosphorylation during G2 and mitosis in fission yeast. We identify substrates of four phosphatases (PP2A-B55, PP2A-B56, CDC14, and PP1), showing that each phosphatase targets a distinct subset of CDK substrate sites. On average, sites dephosphorylated by CDC14 and PP2A-B56 are phosphorylated earlier during G2, followed by sites dephosphorylated by PP1 and then PP2A-B55. This suggests that the identity of the phosphatase impacts the timing of CDK substrate phosphorylation, establishing different phosphorylation thresholds at the G2/M transition. Consistent with this, depletion of PP2A-B55 and CDC14 advances mitotic onset independently of CDK activity regulation, likely due to the earlier phosphorylation of their respective CDK substrates.
]]></description>
<dc:creator>Zeisner, T. U.</dc:creator>
<dc:creator>Auchynnikava, T.</dc:creator>
<dc:creator>Nurse, P.</dc:creator>
<dc:date>2025-02-22</dc:date>
<dc:identifier>doi:10.1101/2025.02.21.639482</dc:identifier>
<dc:title><![CDATA[Phosphatase specificity influences phosphorylation timing of CDK substrates during the cell cycle]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.21.639491v1?rss=1">
<title>
<![CDATA[
Dissecting PADI6 function defines oocyte cytoplasmic lattices as regulatory hubs for fundamental cellular processes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.21.639491v1?rss=1"
</link>
<description><![CDATA[
Oocyte cytoplasmic lattices are critical for early embryo development but their composition and function are not fully understood. Mutations in PADI6, an essential component of cytoplasmic lattices, lead to early embryonic developmental arrest and female infertility. To investigate PADI6 function in mRNA storage, global protein levels, and lattice composition during early mammalian development we used single cell transcriptomics and proteomics methods to study two mouse models. Padi6 null mutation resulted in inhibition of embryonic genome activation, defective maternal mRNA degradation, and disruption to protein storage on the cytoplasmic lattices. Distinct developmental phenotypes were observed with a hypomorphic Padi6 mutation. By developing a powerful single cell proteomic fractionation method, we define the cytoplasmic lattice enriched proteome in which we find essential components of another major oocyte-specific compartment, the endolysosomal vesicular assembly (ELVA), suggesting previously unknown interconnections between them. Our findings highlight a critical scaffolding function of PADI6 and implicate cytoplasmic lattices as regulatory hubs for key processes in the oocyte and early embryo, including translation, respiration and protein degradation.
]]></description>
<dc:creator>Williams, J. P. C.</dc:creator>
<dc:creator>Auchynnikava, T.</dc:creator>
<dc:creator>McCarthy, A.</dc:creator>
<dc:creator>Ojarikre, O. A.</dc:creator>
<dc:creator>Bertran, M. T.</dc:creator>
<dc:creator>Weston, A. E.</dc:creator>
<dc:creator>Leonce, D.</dc:creator>
<dc:creator>Olsen, J.</dc:creator>
<dc:creator>Niakan, K. K.</dc:creator>
<dc:creator>Skehel, M.</dc:creator>
<dc:creator>Turner, J. M. A.</dc:creator>
<dc:creator>Walport, L. J.</dc:creator>
<dc:date>2025-02-22</dc:date>
<dc:identifier>doi:10.1101/2025.02.21.639491</dc:identifier>
<dc:title><![CDATA[Dissecting PADI6 function defines oocyte cytoplasmic lattices as regulatory hubs for fundamental cellular processes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.19.639192v1?rss=1">
<title>
<![CDATA[
3D spatial transcriptomics reveals the molecular domain structure of the mouse olfactory bulb 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.19.639192v1?rss=1"
</link>
<description><![CDATA[
A core organizing principle of the vertebrate brain is its symmetry along multiple axes. However, the precision with which neurons, circuit modules, and brain regions align to these axes remains poorly understood. Here, we used 3D spatial transcriptomics to reconstruct the anatomical and molecular organization of the mouse olfactory bulb. We mapped the positions of nearly one thousand molecularly distinct glomeruli, the structural and functional units of odor processing, revealing highly symmetric organization across hemispheres. Within each bulb, we delineated a curved axis of symmetry that divides pairs of sister glomeruli. Gene expression in the olfactory epithelium predicted glomerular position with near-glomerular resolution. However, glomerular symmetry did not extend to deeper layer mitral and granule cells, suggesting a reorganization from sensory input to cortical output pathways. Our findings provide the first comprehensive map of the olfactory bulb and reveal how its molecular structure is instructed by epithelial gene expression programs.
]]></description>
<dc:creator>Klimpert, N.</dc:creator>
<dc:creator>Kollo, M.</dc:creator>
<dc:creator>Brann, D. H.</dc:creator>
<dc:creator>Tan, C.</dc:creator>
<dc:creator>Barry, D.</dc:creator>
<dc:creator>Ma, Y.</dc:creator>
<dc:creator>Schaefer, A. T.</dc:creator>
<dc:creator>Fleischmann, A.</dc:creator>
<dc:date>2025-02-24</dc:date>
<dc:identifier>doi:10.1101/2025.02.19.639192</dc:identifier>
<dc:title><![CDATA[3D spatial transcriptomics reveals the molecular domain structure of the mouse olfactory bulb]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.25.639898v1?rss=1">
<title>
<![CDATA[
Cryptosporidium modifies intestinal microvilli through an exported virulence factor 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.25.639898v1?rss=1"
</link>
<description><![CDATA[
Cryptosporidium is a common intestinal infection of vertebrates and a significant threat to public health. Within the epithelial layer of the intestine, the parasite invades and replicates. Infected cells are readily detected under the microscope by the presence of elongated microvilli, particularly around the vacuole where the parasite resides. Here, we identify a family of Cryptosporidium virulence factors that are exported into the host cell during infection and localise to the microvilli. We examine the trafficking and function of the most highly expressed family member, MVP1, which appears to control the elongation of microvilli through engagement of host EBP50 and CDC42. Remarkably, this mechanism closely mirrors that of an enteropathogenic Escherichia coli virulence factor, MAP, which is also known to drive host microvilli elongation during infection. This highlights a unique instance where eukaryotic and prokaryotic virulence factors have convergently evolved to modulate host actin structures through a similar mechanism.
]]></description>
<dc:creator>Rodrigues, E.</dc:creator>
<dc:creator>Pallett, M. A.</dc:creator>
<dc:creator>Straker, L. C.</dc:creator>
<dc:creator>Mkandawire, T. T.</dc:creator>
<dc:creator>Sala, K.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:creator>Sateriale, A.</dc:creator>
<dc:date>2025-02-25</dc:date>
<dc:identifier>doi:10.1101/2025.02.25.639898</dc:identifier>
<dc:title><![CDATA[Cryptosporidium modifies intestinal microvilli through an exported virulence factor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.20.639240v1?rss=1">
<title>
<![CDATA[
Dynamic Modeling of Cell Cycle Arrest Through Integrated Single-Cell and Mathematical Modelling Approaches 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.20.639240v1?rss=1"
</link>
<description><![CDATA[
Highly multiplexed imaging assays allow simultaneous quantification of multiple protein and phosphorylation markers, providing a static snapshots of cell types and states. Pseudo-time techniques can transform these static snapshots of unsynchronized cells into dynamic trajectories, enabling the study of dynamic processes such as development trajectories and the cell cycle. Such ordering also enables training of mathematical models on these data, but technical challenges have hitherto made it difficult to integrate multiple experimental conditions, limiting the predictive power and insights these models can generate. In this work, we propose data processing and model training approaches for integrating multiplexed, multi-condition immunofluorescence data with mathematical modelling. We devise training strategies that are applicable to datasets where cells exhibit oscillatory as well as arrested dynamics and use them to train a cell cycle model on a dataset of MCF-10A mammary epithelial exposed to cell-cycle arresting small molecules. We validate the model by investigating predicted growth factor sensitivities and responses to inhibitors of cells at different initial conditions. We anticipate that our framework will generalise to other highly multiplexed measurement techniques such as mass-cytometry, rendering larger bodies of data accessible to dynamic modelling and paving the way to deeper biological insights.

Author SummaryAdvanced imaging techniques allow us to see detailed pictures of different proteins and cell changes. By using computational algorithms, we turn these static pictures into dynamic sequences to understand processes like the cell cycle better. However, combining data from different experiments is difficult and limits how well our models can predict outcomes. This study introduces new ways to process data and train models to handle complex data from various conditions.

The approach is tested by using data from untreated and treated cells to create a model of the cell cycle. This model was then checked for accuracy by seeing how well it could predict how cells respond to growth factors and drugs from different starting points. In the future, this method could be used with other data types, allowing for more detailed and accurate models of cellular behavior.
]]></description>
<dc:creator>Cortes-Rios, J.</dc:creator>
<dc:creator>Rodriguez-Fernandez, M.</dc:creator>
<dc:creator>Sorger, P. K.</dc:creator>
<dc:creator>Fröhlich, F.</dc:creator>
<dc:date>2025-02-26</dc:date>
<dc:identifier>doi:10.1101/2025.02.20.639240</dc:identifier>
<dc:title><![CDATA[Dynamic Modeling of Cell Cycle Arrest Through Integrated Single-Cell and Mathematical Modelling Approaches]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.21.639580v1?rss=1">
<title>
<![CDATA[
Retroelement co-option disrupts the cancer transcriptional programme 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.21.639580v1?rss=1"
</link>
<description><![CDATA[
BackgroundTranscriptional activation of otherwise repressed endogenous retroelements (RTEs) is a hallmark of cancer, shaping tumour progression and immunogenicity by multifaceted, yet incompletely understood mechanisms.

MethodsWe used an extended pan-cancer transcriptome assembly to identify potential effects of RTEs on the genes near or within which they have integrated. These were subsequently verified in test cases by further analysis of transcriptional profiles in cancer patient data, and by in vitro studies involving restoration of gene activity, and proliferation and migration assays in cancer cell lines.

ResultsWe report that cancer-specific transcriptional activation of RTEs causes frequent reduction or loss of gene function. Exonisation and alternative splicing of RTEs creates non-functional RNA and protein isoforms and derepressed RTE promoter activity initiates antisense transcription, both at the expense of the canonical isoforms. Contrary to theoretical expectation, transcriptionally activated RTEs affect genes with established tumour-promoting function, including the common essential RNGTT and the lung cancer-promoting CHRNA5 genes. Furthermore, the disruptive effect of RTE activation on adjacent tumour-promoting genes is associated with slower disease progression in clinical data, whereas experimental restoration of gene activity enhances tumour cell in vitro growth and invasiveness

ConclusionsThese findings underscore the gene-disruptive potential of seemingly innocuous germline RTE integrations, unleashed only by their transcriptional utilisation in cancer. They further suggest that such metastable RTE integrations are co-opted as sensors of the epigenetic and transcriptional changes occurring during cellular transformation and as executors that disrupt the function of tumour-promoting genes.
]]></description>
<dc:creator>Loong, J.</dc:creator>
<dc:creator>Thompson, R.</dc:creator>
<dc:creator>Hall, C.</dc:creator>
<dc:creator>Doglio, L.</dc:creator>
<dc:creator>Pape, J.</dc:creator>
<dc:creator>Kassiotis, G.</dc:creator>
<dc:date>2025-02-27</dc:date>
<dc:identifier>doi:10.1101/2025.02.21.639580</dc:identifier>
<dc:title><![CDATA[Retroelement co-option disrupts the cancer transcriptional programme]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.25.640040v1?rss=1">
<title>
<![CDATA[
Biochemical reconstitution of sister chromatid cohesion establishment during DNA replication 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.25.640040v1?rss=1"
</link>
<description><![CDATA[
Concomitant with DNA replication, the ring-shaped cohesin complex encircles both newly synthesized sister chromatids, enabling their faithful segregation during cell divisions. Our molecular understanding of how cohesin co-entraps both replication products remains incomplete. Here, we reconstitute sister chromatid cohesion establishment using purified budding yeast proteins. Cohesin rings, initially loaded onto template DNA, remain DNA-bound during DNA synthesis and encircle both replication products. Additionally, DNA replication instigates new cohesin recruitment, as a second pathway that generates sister chromatid cohesion. In both scenarios, cohesin often embraces only one of the two replication products, suggestive of a two-step sister chromatid capture mechanism. Sister chromatid co-entrapment occurs independently of replication fork-associated cohesion establishment factors, suggesting a role for the latter during chromatin replication or in facilitating the subsequent cohesin acetylation reaction. Our results make sister chromatid cohesion establishment amenable to direct experimental exploration.
]]></description>
<dc:creator>Minamino, M.</dc:creator>
<dc:creator>Diffley, J.</dc:creator>
<dc:creator>Uhlmann, F.</dc:creator>
<dc:date>2025-02-28</dc:date>
<dc:identifier>doi:10.1101/2025.02.25.640040</dc:identifier>
<dc:title><![CDATA[Biochemical reconstitution of sister chromatid cohesion establishment during DNA replication]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.01.640964v1?rss=1">
<title>
<![CDATA[
Salmonella effector kinase SteC is activated by host-mediated phosphorylation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.01.640964v1?rss=1"
</link>
<description><![CDATA[
The pathogen Salmonella, which causes significant human morbidity and mortality, encodes an effector kinase, SteC, which mediates actin polymerisation and cell migration. Given the minimal nature of its kinase domain, it remains unclear how SteC is catalytically active and how this activity is regulated. Here, we show that SteC is activated following the phosphorylation of the highly conserved S379 residue by a host kinase. Phosphorylation of S379 dramatically increases nucleotide binding affinity of SteC, enabling substrate phosphorylation and promoting actin polymerisation. Further mutational analysis identified the functional role of HD and DGD motifs that likely mimic the HxD and DFG motifs of eukaryotic kinases. Meanwhile, the C-tail of SteC, encompassing amino acids 429-457, is essential for function following translocation from Salmonella, but dispensable for catalysis in vitro. Overall, our findings uncover two previously unappreciated mechanisms that mediate the activity of the only Salmonella effector kinase within the host.

HIGHLIGHTSSteC is phosphorylated at S379 in mammalian cells

SteC phosphorylation is critical for kinase activity and actin polymerisation

Phosphorylation at S379 promotes nucleotide binding

The C-tail of SteC is not required for catalysis but is essential for activity in infected cells
]]></description>
<dc:creator>Pillay, T. D.</dc:creator>
<dc:creator>Lemetais, B.</dc:creator>
<dc:creator>Huber, J.</dc:creator>
<dc:creator>Diaz Del Olmo, I.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Masino, L.</dc:creator>
<dc:creator>Maslen, S.</dc:creator>
<dc:creator>Liu, M.</dc:creator>
<dc:creator>Esposito, D.</dc:creator>
<dc:creator>Hinton, J. C. D.</dc:creator>
<dc:creator>Yu, X.-J.</dc:creator>
<dc:creator>Thurston, T. L. M.</dc:creator>
<dc:creator>Rittinger, K.</dc:creator>
<dc:date>2025-03-02</dc:date>
<dc:identifier>doi:10.1101/2025.03.01.640964</dc:identifier>
<dc:title><![CDATA[Salmonella effector kinase SteC is activated by host-mediated phosphorylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.28.640277v1?rss=1">
<title>
<![CDATA[
A human induced pluripotent stem cell toolbox for studying sex chromosome effects 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.28.640277v1?rss=1"
</link>
<description><![CDATA[
Sex chromosomes shape male (XY) - female (XX) differences in development and disease. These differences can be modelled in vitro by comparing XY and XX human induced pluripotent stem cells (hiPSCs). However, in this system, inter-individual autosomal variation and unstable X-dosage compensation can confound identification of sex chromosomal effects. Here, we utilise sex chromosome loss in XXY fibroblasts to generate XX and XY hiPSCs that are autosomally isogenic and exhibit stable X-dosage compensation. We also create X-monosomic (XO) hiPSCs, to investigate X-Y dosage effects. Using these autosomally isogenic lines, we examine sex differences in pluripotent stem cell expression. Transcriptional differences between XX and XY hiPSCs are surprisingly modest. However, X-haploinsufficiency induces transcriptional deregulation predominantly affecting autosomes. This effect is mediated by Y-genes with broad housekeeping functions that have X-homologues escaping X-inactivation. Our isogenic hiPSC lines provide a resource for exploring sex chromosome effects on development and disease in vitro.
]]></description>
<dc:creator>Meleckyte, R.</dc:creator>
<dc:creator>Varsally, W.</dc:creator>
<dc:creator>Zohren, J.</dc:creator>
<dc:creator>Eriksson, J.</dc:creator>
<dc:creator>Incitti, T.</dc:creator>
<dc:creator>Starnes, L.</dc:creator>
<dc:creator>Pointon, A.</dc:creator>
<dc:creator>Hicks, R.</dc:creator>
<dc:creator>Powell, B. E.</dc:creator>
<dc:creator>Turner, J.</dc:creator>
<dc:date>2025-03-03</dc:date>
<dc:identifier>doi:10.1101/2025.02.28.640277</dc:identifier>
<dc:title><![CDATA[A human induced pluripotent stem cell toolbox for studying sex chromosome effects]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.04.641379v1?rss=1">
<title>
<![CDATA[
Molecular and cellular signatures differentiate Parkinson's disease from Parkinson's disease with dementia 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.04.641379v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) affects millions of people worldwide, and up to 40% of these patients develop dementia, profoundly affecting their quality of life. Whether Parkinsons disease dementia (PDD) simply represents a late stage of PD or constitutes a distinct neurodegenerative process remains unresolved. To clarify this, we generated the largest single nuclear transcriptomic atlas of PD and PDD to date--almost one million nuclei derived from the anterior cingulate cortex and inferior parietal lobule of 64 post-mortem donors. By integrating these data with long-read RNA-seq, we found that the cellular compositions, biological pathways, and molecular profiles diverge substantially between PD and PDD, with limited overlap in differentially expressed genes and pathways. While PD was characterised by widespread upregulation of gene expression programs and robust regional signatures, PDD showed extensive pathway downregulation, loss of cortical regional identity, and significant shifts in transcript usage, including alterations in APP isoforms that may influence pathological amyloid beta accumulation. These findings reveal that PD and PDD represent fundamentally distinct disease states, offering important insights for understanding their underlying mechanisms and will guide the development of targeted therapies and more effective clinical trials.
]]></description>
<dc:creator>Fairbrother-Browne, A.</dc:creator>
<dc:creator>Grant-Peters, M.</dc:creator>
<dc:creator>Brenton, J. W.</dc:creator>
<dc:creator>Nelvagal, H.</dc:creator>
<dc:creator>Reynolds, R. H.</dc:creator>
<dc:creator>Lim, Y. M.</dc:creator>
<dc:creator>Gustavsson, E. K.</dc:creator>
<dc:creator>Macpherson, H.</dc:creator>
<dc:creator>Montgomery, K.</dc:creator>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>Hicks, A. R.</dc:creator>
<dc:creator>Chiraki, N.</dc:creator>
<dc:creator>Curless, T.</dc:creator>
<dc:creator>Real, R.</dc:creator>
<dc:creator>Xenakis, T.</dc:creator>
<dc:creator>Houlden, H.</dc:creator>
<dc:creator>Morris, H. R.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Wood, N. W.</dc:creator>
<dc:creator>Hardy, J.</dc:creator>
<dc:creator>Jaunmuktane, Z.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:date>2025-03-04</dc:date>
<dc:identifier>doi:10.1101/2025.03.04.641379</dc:identifier>
<dc:title><![CDATA[Molecular and cellular signatures differentiate Parkinson's disease from Parkinson's disease with dementia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.02.641068v1?rss=1">
<title>
<![CDATA[
CAR-SPLASH identifies nascent pre-mRNA structures implicated in kinetic coupling and alternative splicing 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.02.641068v1?rss=1"
</link>
<description><![CDATA[
Pre-mRNA splicing is kinetically coupled to transcription as shown by the widespread effects of transcription speed on alternative splicing outcomes. The molecular basis for such kinetic coupling is incompletely understood, but one potential mechanism is through elongation rate dependent alternative folding pathways of the nascent pre-mRNA. To search for RNA structures in nascent pre-mRNA, we modified SPLASH (1) (Sequencing of Psoralen Crosslinked, Ligated And Selected Hybrids) for use with Chromatin Associated RNA. We applied this new method called CAR-SPLASH to cells expressing WT and slow mutant RNA polymerase II and identified > 3000 intramolecular RNA structures of which > 400 are proximal to splice sites. Antisense oligonucleotide (ASO) disruption of several such structures that sequester splice sites has a major impact on alternative splicing outcomes, even though the ASOs do not directly disrupt splice sites. ASO disruption of novel regulatory elements that we designate "RNA kinetic switches", modified alternative splicing of NISCH Exon 18, GAK Exon 7 and MEGF8 Exon 14 in a way that depends on the rate of transcription elongation. We propose that these switches mediate kinetic coupling via effects of transcription speed on folding of nascent RNA structures that modulate alternative splicing, and that many nascent RNA structures can thereby serve as new targets for splice modifying ASOs.

Significance StatementAlternative splicing (AS) of mRNAs is a major regulator of gene expression that is frequently disrupted is disease. AS is affected by the speed of RNA polymerase II that synthesizes mRNA precursors, but how such kinetic coupling works is not well understood. We developed a method, CAR-SPLASH, to capture RNA structures in nascent RNA chains by cross-linking with psoralen. CAR-SPLASH identified novel structural regulatory elements we call "RNA kinetic switches" that control AS in a way that depends on the speed of transcription. RNA kinetic switches are new targets for splice modifying antisense oligonucleotides that could have therapeutic value.
]]></description>
<dc:creator>Shenasa, H.</dc:creator>
<dc:creator>Fong, N.</dc:creator>
<dc:creator>Erickson, B.</dc:creator>
<dc:creator>Iosub, I.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Bentley, D.</dc:creator>
<dc:date>2025-03-06</dc:date>
<dc:identifier>doi:10.1101/2025.03.02.641068</dc:identifier>
<dc:title><![CDATA[CAR-SPLASH identifies nascent pre-mRNA structures implicated in kinetic coupling and alternative splicing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.06.641852v1?rss=1">
<title>
<![CDATA[
Mechanism of DNA-protein crosslink bypass by CMG helicase 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.06.641852v1?rss=1"
</link>
<description><![CDATA[
DNA-protein crosslinks (DPCs) pose significant barriers to DNA replication and genome integrity. Previous studies have established that bypass of DPCs by the eukaryotic replicative helicase, CMG complex, is critical for DPC repair. We investigated the mechanism of leading-strand DPC bypass by CMG using purified components. Our results reveal that CMG can bypass DPCs in the presence of downstream single-stranded DNA, without the aid of other replisome factors. DPC bypass is a slow process, and its efficiency depends on the size and structure of the protein barrier. DPC bypass does not require CMG to interact with the excluded strand or to unfold the protein adduct, implying that the CMG helicase ring opens to navigate past a DPC. Furthermore, opening of no single MCM interface is essential for bypass, suggesting a flexible ring opening mechanism. Our work highlights the remarkable versatility of the CMG complex in navigating replication challenges, ensuring proper replication fork progression and preserving genome stability.
]]></description>
<dc:creator>Kyaw, M. T.</dc:creator>
<dc:creator>Xie, S.</dc:creator>
<dc:creator>de Lamo Peitz, E.</dc:creator>
<dc:creator>Yardimci, H.</dc:creator>
<dc:date>2025-03-06</dc:date>
<dc:identifier>doi:10.1101/2025.03.06.641852</dc:identifier>
<dc:title><![CDATA[Mechanism of DNA-protein crosslink bypass by CMG helicase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.07.642002v1?rss=1">
<title>
<![CDATA[
Predictable Engineering of Signal-DependentCis-Regulatory Elements 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.07.642002v1?rss=1"
</link>
<description><![CDATA[
Cis-regulatory elements (CREs) control how genes respond to external signals, but the principles governing their structure and function remain poorly understood. While differential transcription factor binding is known to regulate gene expression, how CREs integrate the amount and combination of inputs to secure precise spatiotemporal profiles of gene expression remains unclear. Here, we developed a high-throughput combinatorial screening strategy, that we term NeMECiS, to investigate signal- dependent synthetic CREs (synCREs) in differentiating mammalian stem cells. By concatenating fragments of functional CREs from genes that respond to Sonic Hedgehog in the developing vertebrate neural tube, we found that CRE activity follows hierarchical design rules. While individual 200-base-pair fragments showed minimal activity, their combinations generated thousands of functional signal-responsive synCREs, many exceeding the activity of natural sequences. Statistical modelling revealed CRE function can be decomposed into specific quantitative contributions in which sequence fragments combine through a multiplicative rule, tuned by their relative positioning and spacing. These findings provide a predictive framework for CRE redesign, which we used to engineer synthetic CREs that alter the pattern of motor neuron differentiation in neural tissue. These findings establish quantitative principles for engineering synthetic regulatory elements with programmable signal responses to rewire genetic circuits and control stem cell differentiation, providing a basis for understanding developmental gene regulation and designing therapeutic gene expression systems.
]]></description>
<dc:creator>Cornwall-Scoones, J.</dc:creator>
<dc:creator>Benzinger, D.</dc:creator>
<dc:creator>Yu, T.</dc:creator>
<dc:creator>Pezzotta, A.</dc:creator>
<dc:creator>Sagner, A.</dc:creator>
<dc:creator>Gerontogianni, L.</dc:creator>
<dc:creator>Bernadet, S.</dc:creator>
<dc:creator>Finnie, E.</dc:creator>
<dc:creator>Boezio, G. L. M.</dc:creator>
<dc:creator>Stuart, H. T.</dc:creator>
<dc:creator>Melchionda, M.</dc:creator>
<dc:creator>Inge, O. C. K.</dc:creator>
<dc:creator>Dumitrascu, B.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Delas, M. J.</dc:creator>
<dc:date>2025-03-07</dc:date>
<dc:identifier>doi:10.1101/2025.03.07.642002</dc:identifier>
<dc:title><![CDATA[Predictable Engineering of Signal-DependentCis-Regulatory Elements]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.07.641942v1?rss=1">
<title>
<![CDATA[
Mechanical fracturing of the extracellular matrix patterns the vertebrate heart 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.07.641942v1?rss=1"
</link>
<description><![CDATA[
Pattern formation is fundamental to embryonic morphogenesis. In the zebrafish heart, spatially confined single-cell delamination in the ventricle outer curvature initiates trabeculation, a conserved morphogenetic process critical for heart function and embryonic life. Yet, what confines delamination in the ventricle outer curvature remains ill-understood. Contrary to the prevailing notion of patterning through biochemical signals, we now show that mechanical fracturing of the cardiac extracellular matrix (cECM) patterns delamination in the outer curvature. cECM fractures emerge preferentially in the outer curvature, cells delaminate into these fractures and experimental blocking of fractures blocks delamination. These fractures display characteristic signature of mechanical defects and myocardial tissue contractility is sufficient to fracture the cECM, independent of molecular signals, enzymatic activity, or delamination events. Notably, the anisotropic geometry of myocardial tissue generates higher mechanical strain in the outer curvature, thereby locally patterning cECM fractures and delamination. Consequently, cECM fractures evolve in response to dynamic changes in tissue geometry, and experimental manipulation of tissue geometry is sufficient to alter the fracture pattern. Together, our findings underscore mechanical fractures as a morphogenetic strategy, and more generally, corroborate the long-standing but understudied paradigm that tissue form-function can feed back to steer its own patterning.
]]></description>
<dc:creator>Jin Ji, C. C.</dc:creator>
<dc:creator>Santos-Olivan, D.</dc:creator>
<dc:creator>Ramel, M.-C.</dc:creator>
<dc:creator>Sanchez-Posada, J.</dc:creator>
<dc:creator>Paizakis, P.</dc:creator>
<dc:creator>Andrews, T. G.</dc:creator>
<dc:creator>Noel, E. S.</dc:creator>
<dc:creator>Torres-Sanchez, A.</dc:creator>
<dc:creator>Priya, R.</dc:creator>
<dc:date>2025-03-07</dc:date>
<dc:identifier>doi:10.1101/2025.03.07.641942</dc:identifier>
<dc:title><![CDATA[Mechanical fracturing of the extracellular matrix patterns the vertebrate heart]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.05.641116v1?rss=1">
<title>
<![CDATA[
DEK::NUP214 acts as an XPO1-dependent transcriptional activator of essential leukemia genes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.05.641116v1?rss=1"
</link>
<description><![CDATA[
The t(6;9)(p22.3;q34.1) translocation/DEK::NUP214 fusion protein defines a distinct subgroup of younger AML patients classified as a separate disease entity by the World Health Organization. DEK is a nuclear factor with multifunctional roles, including gene regulation, while its fusion partner, NUP214, plays a pivotal role in nuclear export by interacting with transport receptors such as XPO1. However, the precise mechanism by which DEK::NUP214 drives leukemia remains unclear. A comprehensive multi-omics comparison of 57 AML primary samples (including whole genome sequencing, targeted sequencing, transcriptomics, and drug screening with > 500 compounds) revealed that t(6;9) cases display a selective response to XPO1 inhibitors (Selinexor & Eltanexor) and a distinct transcriptomic signature characterized by the overexpression of FOXC1 and HOX genes that are key leukemia mediators. CUT&RUN experiments demonstrated the direct binding of DEK::NUP214 to the promoters of FOXC1 and HOXA/B clusters. Strikingly, the expression of these genes and the binding of DEK::NUP214 to their regulatory regions were selectively reduced upon XPO1 inhibition in t(6;9) cells. Altogether, these results identified a novel function of DEK::NUP214 as an XPO1-dependent transcriptional activator of key leukemia drivers and provide a rationale to explore the use of XPO1 inhibitors in this patient population.
]]></description>
<dc:creator>Kaya, F.</dc:creator>
<dc:creator>Bewicke-Copley, F.</dc:creator>
<dc:creator>Miettinen, J. J.</dc:creator>
<dc:creator>Casado, P.</dc:creator>
<dc:creator>Leddy, E.</dc:creator>
<dc:creator>Deniz, O.</dc:creator>
<dc:creator>Lavallee, V.-P.</dc:creator>
<dc:creator>Philippe, C.</dc:creator>
<dc:creator>Zheng, J.</dc:creator>
<dc:creator>Grebien, F.</dc:creator>
<dc:creator>Khan, N.</dc:creator>
<dc:creator>Krizsan, S.</dc:creator>
<dc:creator>Saad, J.</dc:creator>
<dc:creator>Nolin-Lapalme, A.</dc:creator>
<dc:creator>Lemieux, S.</dc:creator>
<dc:creator>Audemard, E. O.</dc:creator>
<dc:creator>Matthews, J.</dc:creator>
<dc:creator>Grantham, M.</dc:creator>
<dc:creator>Di Bella, D.</dc:creator>
<dc:creator>Wennerberg, K.</dc:creator>
<dc:creator>Parsons, A.</dc:creator>
<dc:creator>Rio-Machin, A.</dc:creator>
<dc:creator>Gribben, J.</dc:creator>
<dc:creator>Cavenagh, J. D.</dc:creator>
<dc:creator>Freeman, S. D.</dc:creator>
<dc:creator>Bodor, C.</dc:creator>
<dc:creator>Sauvageau, G.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Llamas-Sillero, P.</dc:creator>
<dc:creator>Cazier, J.-B.</dc:creator>
<dc:creator>Taussig, D. C.</dc:creator>
<dc:creator>Bonnet, D.</dc:creator>
<dc:creator>Cutillas, P. R.</dc:creator>
<dc:creator>Heckman, C.</dc:creator>
<dc:creator>Fitzgibbon, J.</dc:creator>
<dc:creator>Rouault-Pierre, K.</dc:creator>
<dc:date>2025-03-07</dc:date>
<dc:identifier>doi:10.1101/2025.03.05.641116</dc:identifier>
<dc:title><![CDATA[DEK::NUP214 acts as an XPO1-dependent transcriptional activator of essential leukemia genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.03.640303v1?rss=1">
<title>
<![CDATA[
Granulocyte Derived Resistin Inhibits Monocyte Maturation and Induces Immune Suppression in CMML 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.03.640303v1?rss=1"
</link>
<description><![CDATA[
Chronic myelomonocytic leukaemia (CMML) is a haematological malignancy characterised by overlapping myeloid dysplasia and proliferation with persisting monocytosis. While monocytes are the cardinal malignant cell type in CMML, as a stem cell neoplasm the disease clone comprises most lineages and differentiation stages, including granulocytes. To investigate the pathogenic contribution of granulocytes in CMML maintenance and progression, we performed phenotypic, transcriptomic and functional characterization of CMML granulocytes. Compared with healthy age-matched controls, CMML granulocytes exhibit defective maturation with reduced granularity and phagocytic capacity. Transcriptome analysis revealed activation of pathways linked to proliferation, Myc activity and inflammation. Notably, RETN, which encodes the inflammatory mediator resistin, was upregulated approximately 100-fold in CMML granulocytes; but not differentially expressed in CMML PBMNCs, sorted monocytes, or stem and progenitor cells compared to their healthy counterparts. Accordingly, resistin protein levels were 10-fold higher in plasma from CMML patients and higher plasma resistin levels correlate with poor overall survival and AML-free survival. Remarkably, exposure of healthy monocytes to exogenous recombinant resistin inhibited monocyte maturation and macrophage differentiation. Transcriptome analysis of resistin treated monocytes revealed that resistin induces gene signatures related to immune suppression and myeloid-derived suppressor cell phenotype. We found SEMA4A to be a downstream target of resistin and overexpressed in CMML monocytes. Consistent with known roles for SEMA4A, CMML patients displayed higher percentage of Tregs and elevated Th2/Th1 ratio compared with healthy controls and percentage of Tregs corresponding with associated resistin levels. Furthermore, we demonstrated that resistin directly skews the Th2/Th1 ratio via binding to monocytes. In conclusion, we showed that immature granulocytes in CMML produce high levels of resistin, which contributes to defective monocyte maturation and immune suppression.
]]></description>
<dc:creator>Hull, N. J.</dc:creator>
<dc:creator>Cant, R.</dc:creator>
<dc:creator>Guest, L. A.</dc:creator>
<dc:creator>Wang, Y.-H.</dc:creator>
<dc:creator>Gurashi, K.</dc:creator>
<dc:creator>Paredes, R.</dc:creator>
<dc:creator>Huerga Encabo, H.</dc:creator>
<dc:creator>Lin, C.-C.</dc:creator>
<dc:creator>Tien, H.-F.</dc:creator>
<dc:creator>Sellers, R.</dc:creator>
<dc:creator>Sahoo, S.</dc:creator>
<dc:creator>Bonnet, D.</dc:creator>
<dc:creator>Selimoglu-Buet, D.</dc:creator>
<dc:creator>Solary, E.</dc:creator>
<dc:creator>Wiseman, D. H.</dc:creator>
<dc:creator>Batta, K.</dc:creator>
<dc:date>2025-03-07</dc:date>
<dc:identifier>doi:10.1101/2025.03.03.640303</dc:identifier>
<dc:title><![CDATA[Granulocyte Derived Resistin Inhibits Monocyte Maturation and Induces Immune Suppression in CMML]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.07.642064v1?rss=1">
<title>
<![CDATA[
Engineering Cortical Networks: An Open Platform for Controlled Human Circuit Formation and Synaptic Analysis In vitro 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.07.642064v1?rss=1"
</link>
<description><![CDATA[
Neuronal circuits are complex networks formed by specific neuron connections across brain regions. Understanding their development is key to studying circuit-related dysfunctions in brain diseases. Human-induced pluripotent stem cell (iPSC) models aid in this research but lack precise architecture, limiting insights into neuronal interactions and activity-dependent processes. Microfluidic technologies offer structural control but are restricted by closed systems that hinder 3D integration, scalability, and cell retrieval. To address these limitations, we developed an open cortical network platform integrating iPSC-derived cortical neurons with bioengineering techniques. Using a polydimethylsiloxane (PDMS)-based microgroove topography and a cell plating guide, we created neuronal nodes for controlled circuit assembly. This design enables large-scale functional cortical circuits without physical barriers, allowing optogenetic control of neural activity and flexible network modifications, including cellular composition, neurite directionality, and synapse formation. The open design facilitates neuronal material accessibility, supporting multi-level analyses such as proteomics. This platform serves as a powerful tool for investigating neuronal network development and function, offering new opportunities to study both normal and pathological states, including molecular changes linked to connectivity loss in brain diseases.
]]></description>
<dc:creator>Suklai, P.</dc:creator>
<dc:creator>Minckley, T. F.</dc:creator>
<dc:creator>Hagemann, C.</dc:creator>
<dc:creator>Faber, K.</dc:creator>
<dc:creator>Norkett, R.</dc:creator>
<dc:creator>Guetta, L.</dc:creator>
<dc:creator>OToole, K.</dc:creator>
<dc:creator>Geary, B.</dc:creator>
<dc:creator>Devine, M. J.</dc:creator>
<dc:creator>Serio, A.</dc:creator>
<dc:date>2025-03-10</dc:date>
<dc:identifier>doi:10.1101/2025.03.07.642064</dc:identifier>
<dc:title><![CDATA[Engineering Cortical Networks: An Open Platform for Controlled Human Circuit Formation and Synaptic Analysis In vitro]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.22.644703v1?rss=1">
<title>
<![CDATA[
PhotoNeuro: A compact photodetector for synchronization of visual stimulus presentation during behavioral experiments in neuroscience. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.22.644703v1?rss=1"
</link>
<description><![CDATA[
Presenting visual stimuli in neuroscience experiments often requires the combination of analogue signals that carry information about the visual cue presented on the LCD display. Such signals are often sensed by photodetectors and recorded in analogue to digital converter (ADC) acquisition boards. The use of open-source visual programming languages such as Bonsai is becoming more and more popular. They are often used in combination with other open-source hardware such as Arduino development boards. These microcontroller-based boards can be used to automate behavioural experiments: e.g., actuate valves and motors and acquire analogue signals on their ADC channels. LCDs and other modern display allow fast presentation of arbitrary visual stimuli and are widely used for psychophysics and neuroscience experiments. However, most displays do not provide hardware timestamping options and are intrinsically nonlinear. Solving this limitation often requires a direct recording of the light emitted by the display with a photodiode. Such photodetectors are are often amplified at higher voltages and hard to integrate in most common recording systems that use microcontrollers. The other drawback commonly found by neuroscience researchers in commercial devices is the relatively big footprint that the sensor occupies on the screen which, ideally should be minimised so not to interfere with the stimuli presentation. In this paper we present a small footprint photodetector that can be easily replicated and operates at 5V making it suitable to use with common development boards and the visual programming language Bonsai that is commonly used for experiment creation and control. Additionally, we share a version that includes four photodiodes in small area (400 mm2).

Specifications table

O_TBL View this table:
org.highwire.dtl.DTLVardef@7bafcforg.highwire.dtl.DTLVardef@5353ccorg.highwire.dtl.DTLVardef@a29f47org.highwire.dtl.DTLVardef@86f5d2org.highwire.dtl.DTLVardef@74da5f_HPS_FORMAT_FIGEXP  M_TBL C_TBL
]]></description>
<dc:creator>Cano Ferrer, X.</dc:creator>
<dc:creator>Moglie, M. J.</dc:creator>
<dc:creator>Konstantinou, G.</dc:creator>
<dc:creator>Blot, A.</dc:creator>
<dc:creator>Bianchini, G.</dc:creator>
<dc:creator>Imbert, A.</dc:creator>
<dc:creator>Znamenskiy, P.</dc:creator>
<dc:creator>Iacaruso, M. F.</dc:creator>
<dc:date>2025-03-24</dc:date>
<dc:identifier>doi:10.1101/2025.03.22.644703</dc:identifier>
<dc:title><![CDATA[PhotoNeuro: A compact photodetector for synchronization of visual stimulus presentation during behavioral experiments in neuroscience.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.25.645230v1?rss=1">
<title>
<![CDATA[
Mapping and quantifying nascent transcript start sites using TT-TSS-seq 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.25.645230v1?rss=1"
</link>
<description><![CDATA[
Transcription initiation is a highly dynamic and tightly regulated process involving the coordinated action of transcription factors, chromatin remodelers, and RNA polymerase which determine where and when transcription begins. Accurately mapping and quantifying transcription start sites (TSSs) from nascently transcribed RNAs remains a key area of interest, as it provides critical insights into transcription dynamics. Here, we combined transient transcriptome sequencing with transcription start site sequencing (TT-TSS-seq) to accurately map and quantify transcription initiation sites from nascent transcripts. Since transient metabolic labelling yields low-input RNA, we optimized the TSS-seq protocol to enhance sensitivity and accuracy. Specifically, we refined enzymatic reactions for decapping and RNA ligation and incorporated 5 oligonucleotides containing unique molecular identifiers (UMIs) and barcodes to enable accurate quantification and sample multiplexing. The TT-TSS-seq approach detected transcription initiation of unstable transcripts, such as enhancer RNAs. Moreover, we identified that a large fraction of genes use multiple transcription initiation sites, yet often produce only a single stable transcript. Overall, TT-TSS-seq provides precise mapping and quantification of transcription initiation sites, offering new insights into transcriptional dynamics and expanding the toolkit for studying gene regulation.
]]></description>
<dc:creator>Elgood Hunt, E.</dc:creator>
<dc:creator>Vivori, C.</dc:creator>
<dc:creator>van Werven, F.</dc:creator>
<dc:date>2025-03-27</dc:date>
<dc:identifier>doi:10.1101/2025.03.25.645230</dc:identifier>
<dc:title><![CDATA[Mapping and quantifying nascent transcript start sites using TT-TSS-seq]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.25.645220v1?rss=1">
<title>
<![CDATA[
DNA supercoiling impacts alternative transcription start site selection in yeast 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.25.645220v1?rss=1"
</link>
<description><![CDATA[
Most genes are transcribed from multiple transcription start sites (TSSs), defined as alternative TSSs, which are highly regulated and can lead to various gene regulatory outcomes including changes in translation efficiency and protein isoform expression. Transcription factors and chromatin regulators control alternative TSS selection. DNA supercoiling affects multiple aspects of transcription including transcription initiation; however, its regulatory effect on genes with multiple TSSs is not known. Here, we investigated how DNA supercoiling impacts alternative TSS usage in Saccharomyces cerevisiae. We depleted topoisomerases during early meiosis, where alternative TSS usage is prevalent, and applied an improved TSS sequencing protocol. We show that supercoiling affects alternative TSS usage at almost 600 genes. Increased alternative and aberrant TSS usage was observed near and within open reading frames, likely resulting from transcription-induced supercoiling originating from upstream alternative TSSs. DNA supercoiling had the greatest impact on genes with a dominant alternative TSS, significant spacing between alternative TSSs, and greater overall gene length. Our results establish that DNA supercoiling release during transcription is critical for correct TSS selection.
]]></description>
<dc:creator>Elgood Hunt, E.</dc:creator>
<dc:creator>Vivori, C.</dc:creator>
<dc:creator>van Werven, F.</dc:creator>
<dc:date>2025-03-26</dc:date>
<dc:identifier>doi:10.1101/2025.03.25.645220</dc:identifier>
<dc:title><![CDATA[DNA supercoiling impacts alternative transcription start site selection in yeast]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.06.641848v1?rss=1">
<title>
<![CDATA[
PRMT activity promotes global 3' UTR shortening in proliferating cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.06.641848v1?rss=1"
</link>
<description><![CDATA[
Protein methyltransferase (PRMT)-catalysed arginine methylation is a widespread post-translational modification that regulates numerous RNA-binding proteins and frequently becomes dysregulated in cancer. While PRMT inhibitors have shown promise as an anti-cancer strategy, greater understanding of the downstream pathways linking arginine methylation to tumour-promoting phenotypes is needed to improve patient stratification and develop more effective therapeutic approaches. Here, we reveal arginine methylation as a critical regulator of alternative polyadenylation (APA) patterns that are fundamental to tumour progression. 3' RNA-sequencing assays uncover a rapid and global shift toward longer 3' UTR isoforms upon dual (symmetric and asymmetric) methylation (DMAi), impacting a broad range of cellular proliferation and signalling genes. Arginine methylation is required for sustaining proximal poly(A) site usage under high proliferative demand, as DMAi treatment blocks use of such sites in activated T cells, various cancer cell lines and patient-derived lung organoids. DMAi also counteracts the 3' UTR shortening caused by reduced CFIM25 expression, which normally promotes oncogenic isoforms. DMAi treatment affects APA in many of the same mRNAs as impaired cleavage and polyadenylation activity, and these mRNAs contain characteristic signatures such as high GC-content and long 3 UTRs. This systematic impact of PRMT activity on APA regulation broadens the potential utility of PRMT inhibitors as therapeutic agents for both cancer and immune-related diseases.
]]></description>
<dc:creator>Griffith, L.</dc:creator>
<dc:creator>Capitanchik, C.</dc:creator>
<dc:creator>Moore, S.</dc:creator>
<dc:creator>Farcas, A.</dc:creator>
<dc:creator>Gwynne, M.</dc:creator>
<dc:creator>Pedna, M.</dc:creator>
<dc:creator>Jones, D. M.</dc:creator>
<dc:creator>Chakrabarti, A. M.</dc:creator>
<dc:creator>Lagos, D.</dc:creator>
<dc:creator>Urosevic, J.</dc:creator>
<dc:creator>Lynch, J. T.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:date>2025-03-10</dc:date>
<dc:identifier>doi:10.1101/2025.03.06.641848</dc:identifier>
<dc:title><![CDATA[PRMT activity promotes global 3' UTR shortening in proliferating cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.14.643248v1?rss=1">
<title>
<![CDATA[
The evolution of gene regulation in mammalian cerebellum development 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.14.643248v1?rss=1"
</link>
<description><![CDATA[
Gene regulatory changes are considered major drivers of evolutionary innovations, including the cerebellums expansion during human evolution, yet they remain largely unexplored. In this study, we combined single-nucleus measurements of gene expression and chromatin accessibility from six mammals (human, bonobo, macaque, marmoset, mouse, and opossum) to uncover conserved and diverged regulatory networks in cerebellum development. We identified core regulators of cell identity and developed sequence-based models that revealed conserved regulatory codes. By predicting chromatin accessibility across 240 mammalian species, we reconstructed the evolutionary histories of human cis-regulatory elements, identifying sets associated with positive selection and gene expression changes, including the recent gain of THRB expression in cerebellar progenitor cells. Collectively, our work reveals the shared and mammalian lineage-specific regulatory programs governing cerebellum development.
]]></description>
<dc:creator>Sarropoulos, I.</dc:creator>
<dc:creator>Sepp, M.</dc:creator>
<dc:creator>Yamada, T.</dc:creator>
<dc:creator>Schaefer, P. S. L.</dc:creator>
<dc:creator>Trost, N.</dc:creator>
<dc:creator>Schmidt, J.</dc:creator>
<dc:creator>Schneider, C.</dc:creator>
<dc:creator>Drummer, C.</dc:creator>
<dc:creator>Missbach, S.</dc:creator>
<dc:creator>Taskiran, I. I.</dc:creator>
<dc:creator>Hecker, N.</dc:creator>
<dc:creator>Bravo Gonzalez-Blas, C.</dc:creator>
<dc:creator>Kempynck, N.</dc:creator>
<dc:creator>Froemel, R.</dc:creator>
<dc:creator>Joshi, P.</dc:creator>
<dc:creator>Leushkin, E.</dc:creator>
<dc:creator>Arnskoetter, F.</dc:creator>
<dc:creator>Leiss, K.</dc:creator>
<dc:creator>Okonechnikov, K.</dc:creator>
<dc:creator>Lisgo, S.</dc:creator>
<dc:creator>Palkovits, M.</dc:creator>
<dc:creator>Paabo, S.</dc:creator>
<dc:creator>Cardoso-Moreira, M.</dc:creator>
<dc:creator>Kutscher, L. M.</dc:creator>
<dc:creator>Behr, R.</dc:creator>
<dc:creator>Pfister, S. M.</dc:creator>
<dc:creator>Aerts, S.</dc:creator>
<dc:creator>Kaessmann, H.</dc:creator>
<dc:date>2025-03-16</dc:date>
<dc:identifier>doi:10.1101/2025.03.14.643248</dc:identifier>
<dc:title><![CDATA[The evolution of gene regulation in mammalian cerebellum development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.14.643230v1?rss=1">
<title>
<![CDATA[
Autologous human iPSC-derived Alveolus-on-Chip reveals early pathological events of M. tuberculosis infection 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.14.643230v1?rss=1"
</link>
<description><![CDATA[
Immunocompetent and experimentally accessible alveolar systems to study human respiratory diseases are lacking. Here, we developed a single donor human induced pluripotent stem cell (iPSC)-derived Lung-on-Chip (iLoC) containing Type II and I alveolar epithelial cells, vascular endothelial cells, and macrophages in a microfluidic device that mimic lung 3D mechanical stretching and air-liquid interface. Imaging and scRNA-seq analysis revealed that the iLoC recapitulated cellular profiles present in the human distal lung. Infection of the iLoC with the human pathogen Mycobacterium tuberculosis (Mtb) showed that both macrophages and epithelial cells were infected and showed limited bacterial replication. Stochastically, large macrophage clusters containing necrotic core-like structure and Mtb replication were observed. A genetically engineered autophagy deficient iLoC revealed that after Mtb infection, macrophage necrosis was higher upon ATG14 deficiency without bacterial replication. Altogether, we report an autologous, genetically tractable human alveolar model to study lung diseases and therapies.
]]></description>
<dc:creator>Luk, C. H.</dc:creator>
<dc:creator>Conway, G.</dc:creator>
<dc:creator>Goh, K. J.</dc:creator>
<dc:creator>Fearns, A.</dc:creator>
<dc:creator>Rodriguez-Hernandez, I.</dc:creator>
<dc:creator>D'Antuono, R.</dc:creator>
<dc:creator>Day, N. J.</dc:creator>
<dc:creator>Athanasiadi, N.</dc:creator>
<dc:creator>Pellegrino, E.</dc:creator>
<dc:creator>Hobi, N.</dc:creator>
<dc:creator>Stucki, J.</dc:creator>
<dc:creator>Gutierrez, M. G.</dc:creator>
<dc:date>2025-03-17</dc:date>
<dc:identifier>doi:10.1101/2025.03.14.643230</dc:identifier>
<dc:title><![CDATA[Autologous human iPSC-derived Alveolus-on-Chip reveals early pathological events of M. tuberculosis infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.23.644738v1?rss=1">
<title>
<![CDATA[
Population activity of mossy fibre axon input to the cerebellar cortex during behaviours 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.23.644738v1?rss=1"
</link>
<description><![CDATA[
The cerebellum gathers information from the neocortex via the cortico-ponto-cerebellar pathway, and forms sensorimotor associations for predicting movements and their sensory consequences. However, little is known about the functional properties of this major input to the cerebellar cortex. Recordings from individual cerebellar mossy fibre axons (MFAs) have shown that they convey sensory and motor information, but nothing is known about their population code. Here, we report that the population activity of pontine MFAs is heterogeneous, high-dimensional and that different subpopulations of MFAs are active during quiet and active behavioural states. Population activity occupied a substantial fraction of the state space and some MFAs are particularly informative about behaviour. Surprisingly, positively and negatively modulated MFAs are intermingled, suggesting granule cells integrate opposite-signed inputs to generate mixed bidirectional sensorimotor representations. Our results establish that neocortex and cerebellum can communicate with a low redundancy, high capacity, bidirectional population code, which is well-suited for forming sensorimotor associations.

Highlights[bullet] Population activity of main mossy fibre axonal input to cerebellar cortex.
[bullet]Ponto-cerebellar code is high dimensional during behaviours.
[bullet]Behavioural information conveyed by bidirectional population code.
[bullet]Modelling predicts heterogeneous response properties of granule cells.
]]></description>
<dc:creator>Ros, H.</dc:creator>
<dc:creator>Xie, Y.</dc:creator>
<dc:creator>Sadeh, S.</dc:creator>
<dc:creator>Silver, R. A.</dc:creator>
<dc:date>2025-03-24</dc:date>
<dc:identifier>doi:10.1101/2025.03.23.644738</dc:identifier>
<dc:title><![CDATA[Population activity of mossy fibre axon input to the cerebellar cortex during behaviours]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.27.643001v1?rss=1">
<title>
<![CDATA[
Two distinct cytokine response clusters identified in healthcare workers with apparent resistance to infection with Mycobacterium tuberculosis despite sustained occupational exposure 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.27.643001v1?rss=1"
</link>
<description><![CDATA[
IntroductionEvidence exists that some individuals resist ("Resisters") Mycobacterium tuberculosis (Mtb) infection despite sustained exposure, but the protective mechanisms involved are not fully understood. We investigated immune responses induced by ex vivo stimulation of peripheral blood mononuclear cells (PBMC) with live Mtb in Resisters compared to those with latent TB infection (LTBI).

MethodsHIV-uninfected healthcare workers working in high TB exposure healthcare facilities for over 5 years were screened for Mtb sensitization using the interferon-gamma release assay (IGRA) and the tuberculin skin test (TST). We identified Resisters (TST<10mm and IGRA<0.35 IU/mL, n=129) and those with LTBI (TST[&ge;]10mm and IGRA[&ge;]0.35 IU/mL n=145). We selected a subset of  extreme resisters (TST=0mm and IGRA<0.2 IU/mL; n=26) and  extreme LTBI (TST[&ge;]15mm and IGRA[&ge;]1 IU/mL; n=24) for these analyses. Blood was collected and PBMC isolated and cultured with live H37Rv Mtb for 18 hours. Supernatants were collected and used for measuring 65 secreted analytes by Luminex. We also evaluated cell-associated cytokine expression by CD4 T cells and monocytes in these participants using flow cytometry.

ResultsUsing the feature selection in R, we identified a set of 4 cytokines: IL-17A, MCP-1, IL-8, and MDC, which collectively classified extreme Resisters from extreme LTBI with an area-under-the-curve (AUC) of 0.67 (0.54-0.85). Focusing only on the extreme Resisters, and using hierarchical clustering, participants in this group segregated into two main clusters (Resister_c1 and Resister_c2). Further analyses identified 37 cytokines that were significantly higher and 15 cytokines that were lower in Resister_c2 compared with Resister_c1. A set of 5 cytokines (TRAIL, MIP-1{beta}, Fractalkine, GRO- and IL-1) collectively classified these two clusters with an AUC of 1 (1-1). CD4 T cell and monocyte responses to Mtb did not significantly differ between the two clusters.

ConclusionsMtb-specific immune responses segregate extreme Resisters into two distinct clusters in individuals with apparent resistance to Mtb infection and may allow discrimination of individuals with true resistance from those with infection but have alternative immune responses not detected by IGRA and TST.
]]></description>
<dc:creator>Balfour, A.</dc:creator>
<dc:creator>Masina, N.</dc:creator>
<dc:creator>Queiroz, A.</dc:creator>
<dc:creator>Andrade, B.</dc:creator>
<dc:creator>Fukutani, E.</dc:creator>
<dc:creator>Bekiswa, A.</dc:creator>
<dc:creator>Lewinsohn, D.</dc:creator>
<dc:creator>Lewinsohn, D.</dc:creator>
<dc:creator>Goliath, R.</dc:creator>
<dc:creator>Wilkinson, K. A.</dc:creator>
<dc:creator>Schutz, C.</dc:creator>
<dc:creator>Meintjes, G.</dc:creator>
<dc:creator>Shey, M. S.</dc:creator>
<dc:date>2025-03-28</dc:date>
<dc:identifier>doi:10.1101/2025.03.27.643001</dc:identifier>
<dc:title><![CDATA[Two distinct cytokine response clusters identified in healthcare workers with apparent resistance to infection with Mycobacterium tuberculosis despite sustained occupational exposure]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.25.645352v1?rss=1">
<title>
<![CDATA[
Structural Basis of CSN-mediated SCF Deneddylation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.25.645352v1?rss=1"
</link>
<description><![CDATA[
Cullin-RING ligases (CRLs) are the largest family of E3 ligases, with their ubiquitination activity dynamically regulated by neddylation and deneddylation by the COP9 signalosome (CSN). CSN-mediated deneddylation not only deactivates CRLs but also enables substrate receptor exchange, fine-tuning CRL specificity. CSN has emerged as a promising drug target, yet the structural basis underlying its catalytic mechanism remains insufficiently understood. To address this, we used cryo-electron microscopy (cryo-EM) analysis of CSN-CRL1 (SCF) complexes to uncover distinct functional states, capturing key intermediates of the deneddylation cycle. The earliest state represents an initial docking step in which CSN remains autoinhibited. In contrast, the catalytic intermediate reveals the fully engaged state, with the CSN5 Ins-1 loop, RBX1 RING, and neddylated Cullin WHB domains repositioned for isopeptide bond cleavage. We further resolve four dissociation intermediates that define the stepwise release of CSN from its deneddylated product, highlighting a central role for RBX1RING in stabilising key interactions throughout this process. Additionally, our structures reveal the previously uncharacterised position of CSNAP, which integrates into the CSN scaffold at a groove formed by CSN3 and CSN8. Together, our study provides a comprehensive mechanistic model of CSN function, linking CRL recognition, catalytic activation, and stepwise disengagement. These insights lay the groundwork for the rational design of CSN inhibitors, offering new opportunities to modulate CRL activity for therapeutic applications.

Key TakeawaysO_LIHigh-resolution cryo-EM structures capture key intermediates of the CSN-mediated SCF deneddylation cycle
C_LIO_LIStructural identification of CSNAP places it within a previously uncharacterized groove at the interface of CSN3 and CSN8.
C_LIO_LIPre-catalytic and catalytic states define the conformational transitions of CSN5Ins-1, RBX1RING, and N8WHB that enable isopeptide bond cleavage.
C_LIO_LIFour dissociation intermediates reveal a sequential pathway for CSN disengagement, with RBX1RING playing a central role in stabilising key interactions.
C_LI
]]></description>
<dc:creator>Enchev, R. I.</dc:creator>
<dc:creator>Ding, S.</dc:creator>
<dc:creator>Clapperton, J. A.</dc:creator>
<dc:creator>Maeots, M.-E.</dc:creator>
<dc:creator>Kunzelmann, S.</dc:creator>
<dc:creator>Shaaban, M.</dc:creator>
<dc:date>2025-03-26</dc:date>
<dc:identifier>doi:10.1101/2025.03.25.645352</dc:identifier>
<dc:title><![CDATA[Structural Basis of CSN-mediated SCF Deneddylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.27.645708v1?rss=1">
<title>
<![CDATA[
The ubiquitin E3 ligase Huwe1 facilitates viral and self RNA sensing by RIG-I-like receptors 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.27.645708v1?rss=1"
</link>
<description><![CDATA[
RIG-I-like receptors (RLRs) are cytoplasmic RNA sensors that promote type I and type III interferon (IFN) production in response to RNA ligands of viral or endogenous origin. The RLR pathway is tightly regulated by dynamic post-translational modifications, including ubiquitination. Huwe1 is a HECT domain-containing giant ubiquitin E3 ligase that has not been implicated in the RLR or IFN pathway. Here, we investigated whether Huwe1 is required for type I IFN induction downstream of RLRs. We demonstrate that loss of Huwe1 severely attenuates the expression of IFN-{beta}, IFN-{lambda}1 and IFN-stimulated genes (ISGs) in ADAR1-deficient human cells and primary murine bone-marrow derived macrophages, in which unedited self RNAs that serve as RLR ligands accumulate. In addition, depletion of Huwe1 reduces the induction of type I and III IFNs upon transfection with synthetic viral RNA mimetics or infection with a picornavirus. Using proteomics, we identified several putative Huwe1 substrates, which include key components of the RLR pathway (MAVS, TRAFs). We demonstrate that these substrates interact with Huwe1 and that Huwe1 is essential for the activity of TRAF5 in type I IFN induction. Collectively, our results put Huwe1 on the map as an important ubiquitin E3 ligase in the RLR pathway and provide new insights into ubiquitin-dependent regulation of cell-intrinsic antiviral immune pathways.
]]></description>
<dc:creator>Oosenbrug, T.</dc:creator>
<dc:creator>Gravekamp, D.</dc:creator>
<dc:creator>Pinto-Fernandez, A.</dc:creator>
<dc:creator>Crotta, S.</dc:creator>
<dc:creator>Finsterbusch, K.</dc:creator>
<dc:creator>van der Kooij, S. B.</dc:creator>
<dc:creator>ter Haar, L. R.</dc:creator>
<dc:creator>Asschert, Y.</dc:creator>
<dc:creator>Kleian, G. F. E.</dc:creator>
<dc:creator>Kessler, B. M.</dc:creator>
<dc:creator>Wack, A.</dc:creator>
<dc:creator>van der Veen, A. G.</dc:creator>
<dc:date>2025-04-01</dc:date>
<dc:identifier>doi:10.1101/2025.03.27.645708</dc:identifier>
<dc:title><![CDATA[The ubiquitin E3 ligase Huwe1 facilitates viral and self RNA sensing by RIG-I-like receptors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.04.02.645514v1?rss=1">
<title>
<![CDATA[
An intercellular metabolic relay for brain sparingin Drosophila 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.04.02.645514v1?rss=1"
</link>
<description><![CDATA[
Brain sparing protects growth of the central nervous system (CNS) at the expense of other developing organs during nutrient restriction. This survival strategy is conserved from Drosophila to mammals but little is known about its underlying metabolic mechanisms. Here, we show that CNS uptake and catabolism of circulating glutamine is essential for neural stem cell proliferation during brain sparing but not normal brain development. Glutamine is imported into perineurial glia of the blood-brain-barrier, which upregulate glutaminase (Gls) and vesicular glutamate transporter 1 (VGlut1) to promote glutaminolysis and glutamate secretion. Neural stem cells then import released glutamate via excitatory amino acid transporter 1 (Eaat1) and maintain it at homeostatic levels via glutamine synthetase 1 (Gs1). Glutamine nitrogen contributes surprisingly little to newly synthesized DNA or protein in neural stem cells but its carbon fuels the oxidative tricarboxylic acid cycle. These findings identify an intercellular metabolic relay specifically required to sustain the proliferation of neural stem cells during brain sparing.
]]></description>
<dc:creator>Franchet, A.</dc:creator>
<dc:creator>Jin, Y.</dc:creator>
<dc:creator>Newell, C. L.</dc:creator>
<dc:creator>Girard, V.</dc:creator>
<dc:creator>Maniere, G.</dc:creator>
<dc:creator>Grosjean, Y.</dc:creator>
<dc:creator>Barrington, C.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Gilmore, I. S.</dc:creator>
<dc:creator>Gould, A. P.</dc:creator>
<dc:date>2025-04-03</dc:date>
<dc:identifier>doi:10.1101/2025.04.02.645514</dc:identifier>
<dc:title><![CDATA[An intercellular metabolic relay for brain sparingin Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.24.614724v1?rss=1">
<title>
<![CDATA[
Why do we have so many excitatory neurons? 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.24.614724v1?rss=1"
</link>
<description><![CDATA[
The emerging electron microscopy connectome datasets provides connectivity maps of the brains at single cell resolution, enabling us to estimate various network statistics, such as connectedness. We desire the ability to assess how the functional complexity of these networks depends on these network statistics. To this end, we developed an analysis pipeline and a statistic, XORness, which quantifies the functional complexity of these networks with varying network statistics. We illustrate that actual connectomes have high XORness, as do generated connectomes with the same network statistics, suggesting a normative role for functional complexity in guiding the evolution of connectomes, and providing clues to guide the development of artificial neural networks.
]]></description>
<dc:creator>Wang, Q.</dc:creator>
<dc:creator>Cardona, A.</dc:creator>
<dc:creator>Zlatic, M.</dc:creator>
<dc:creator>Vogelstein, J.</dc:creator>
<dc:creator>Priebe, C.</dc:creator>
<dc:date>2024-09-24</dc:date>
<dc:identifier>doi:10.1101/2024.09.24.614724</dc:identifier>
<dc:title><![CDATA[Why do we have so many excitatory neurons?]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.04.08.647770v1?rss=1">
<title>
<![CDATA[
VP-CLEM-Kit: An accessible pipeline for visual proteomics using super resolution volume correlative light and electron microscopy (SR-vCLEM) 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.04.08.647770v1?rss=1"
</link>
<description><![CDATA[
Visual proteomics (VP) aims to allow researchers to visualise, measure and analyse proteins in the context of cell and tissue structure in health and disease. VP is becoming a reality through technological advances across several domains, including in situ structural biology and correlative light and electron microscopy (CLEM). However, widespread adoption remains limited due to the complexity and cost of the various VP approaches reported to date. Here we present the VP-CLEM-Kit, a disruptive cost-effective pipeline for super resolution volume CLEM (SR-vCLEM) that can be implemented with minimal advanced electron microscopy expertise and equipment, making it accessible to light microscopy facilities and research labs. SR-vCLEM is based on in-resin fluorescence (IRF), where fluorophores are preserved through processing into resin. The easyIRF protocol reported here reduces the requirement for complex costly sample preparation equipment and toxic chemicals compared to standard IRF protocols. easyIRF blocks are cut into ultrathin sections that are imaged using  tomoSTORM, a new modular and cost-effective openFrame-based light microscope controlled by the open-source software package Micro-Manager that provides serial single molecule localisation microscopy in array tomography format. Sections are then post-stained and imaged using a tabletop scanning electron microscope controlled by open-source SBEMimage software to run in array tomography format. We demonstrate the potential of the VP-CLEM-Kit by imaging organelle reporters in human cell lines and stem cell derived neurons, fluorescently labelled protein in neurons, and immunolabelled cells in human kidney biopsy tissue from transplant patients. The VP-CLEM-Kit delivers a [~]5-fold improvement in resolution at a [~]7-fold lower cost, and thus provides new technical capability as well as a blueprint for more equitable access to advanced imaging workflows.
]]></description>
<dc:creator>Lumkwana, D.</dc:creator>
<dc:creator>Lightley, J.</dc:creator>
<dc:creator>Vesga, A. G.</dc:creator>
<dc:creator>de Folter, J.</dc:creator>
<dc:creator>Domart, M.-C.</dc:creator>
<dc:creator>Maclachlan, C.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>Peddie, C.</dc:creator>
<dc:creator>Burrell, A.</dc:creator>
<dc:creator>Yoshimura, A.</dc:creator>
<dc:creator>Mangali, A.</dc:creator>
<dc:creator>Vahrjmeijer, N.</dc:creator>
<dc:creator>Bhaga, M.</dc:creator>
<dc:creator>Smit, C.</dc:creator>
<dc:creator>Titze, B.</dc:creator>
<dc:creator>Horrocks, M. H.</dc:creator>
<dc:creator>Cobra Straker, L.</dc:creator>
<dc:creator>Garcia, E.</dc:creator>
<dc:creator>Jones, M. L.</dc:creator>
<dc:creator>Mclean, A.</dc:creator>
<dc:creator>Roufosse, C.</dc:creator>
<dc:creator>Loos, B.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Strange, A.</dc:creator>
<dc:creator>Henriques, R.</dc:creator>
<dc:creator>French, P. M.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:date>2025-04-10</dc:date>
<dc:identifier>doi:10.1101/2025.04.08.647770</dc:identifier>
<dc:title><![CDATA[VP-CLEM-Kit: An accessible pipeline for visual proteomics using super resolution volume correlative light and electron microscopy (SR-vCLEM)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.04.21.649820v1?rss=1">
<title>
<![CDATA[
Chronobot: Deep learning guided time-resolved cryo-EM captures molecular choreography of RecA in homology search 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.04.21.649820v1?rss=1"
</link>
<description><![CDATA[
The function of proteins and other biological macromolecules is regulated by conformational dynamics1. Many functional changes take place on millisecond timescales which cannot be experimentally captured by manual sample preparation for cryo-EM2. Here we introduce Chronobot, a robust, data-driven platform enabling reproducible, time-resolved cryo-EM sample preparation to visualize these transient intermediates. We quantify ice thickness, an important precondition and thus a reliable predictor of 3D reconstruction quality, using two complementary methods: a bespoke deep learning model analysing high-speed camera videos of the grid just before vitrification, and detailed ice thickness quantifications of outputs from common TEM screening workflows like the EPU software. Combining these methods enables rapid optimisation, resulting in an 11-fold improvement in cryo-EM sample quality compared to our previously reported workflow3.

To demonstrate the Chronobot in capturing transient reaction intermediates visualised through cryo-EM and single particle analysis we focused on RecA homology search. The RecA family of recombinases perform the essential task of rapidly scanning for homologous dsDNA sequences to initiate homologous recombination. The dynamics of these RecA-dsDNA interactions occur on millisecond timescales, limiting structural insights4. We capture time-resolved homology search intermediates at 250 milliseconds. These structures reveal the involvement of the secondary DNA binding site in initial capture of dsDNA before homology sampling occurs. We also observe three-strand homology sampling intermediates, where the homologous strand is not fully displaced, and homology is not stably bound. Our results suggest a model of how RecA-family recombinases function in early homologous recombination, by coordinating the incoming DNA between RecAs various DNA binding sites depending on the stage of homology search and the presence of suUicient homology. We anticipate the Chronobot method to be broadly applicable to processes which cannot be captured by manual sample preparation methods. In addition, by leveraging AI inference, our rapid user feedback mechanisms allow for per-sample optimisation of grid conditions, increasing the likelihood of success and reducing the sample requirements of each time-resolved experiment.
]]></description>
<dc:creator>Mäeots, M.-E.</dc:creator>
<dc:creator>Tupin, S.</dc:creator>
<dc:creator>Esfahani, N. M. M.</dc:creator>
<dc:creator>Molina, J. B. R.</dc:creator>
<dc:creator>Clapperton, J. A.</dc:creator>
<dc:creator>Amin, A.</dc:creator>
<dc:creator>Imbert, A.</dc:creator>
<dc:creator>Enchev, R. I.</dc:creator>
<dc:date>2025-04-22</dc:date>
<dc:identifier>doi:10.1101/2025.04.21.649820</dc:identifier>
<dc:title><![CDATA[Chronobot: Deep learning guided time-resolved cryo-EM captures molecular choreography of RecA in homology search]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.04.24.650439v1?rss=1">
<title>
<![CDATA[
Structure-Function Mapping of Olfactory Bulb Circuits with Synchrotron X-ray Nanotomography 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.04.24.650439v1?rss=1"
</link>
<description><![CDATA[
Information is routed between brain areas via parallel streams. Neurons may share common inputs yet convey distinct information to different downstream targets. Here, we leverage the anatomical organisation of the mouse olfactory bulb (OB), where dozens of projection neurons (mitral and tufted cells, M/TCs) affiliate with a single input unit, a glomerulus1-3. To link functional properties of M/TCs to their anatomical glomerular association at scale, we combine in vivo two-photon (2P) imaging with synchrotron {micro}CT4-6 anatomical analysis and targeted X- ray nano-holotomography (XNH)7,8. Improving XNH resolution for mm3 volumes enables us to reliably identify subcellular features, automatically segment >80,000 cell nuclei in individual experiments, and delineate several hundred functionally imaged projection neurons and their detailed morphology, including up to 20 M/TCs per individual glomerulus ("sister" cells). In over 2400 sister cell pairs, we consistently find that odour response amplitudes to a panel of 47 monomolecular odours are conserved between sister cells, with, however, distinct responses to individual odours. Responses correlated with anatomical features such as cell body position and lateral dendritic arborisation. Thus, sister cells neither simply relay glomerular inputs nor are they dominated by network activity. Instead, they show a "balanced diversity" in their responses, enabling efficient encoding of odour stimuli whilst retaining the overall structure of odour space. Thus, synchrotron X-ray tomography can reliably link subcellular anatomy to function in a non-destructive way across the mm3 scale. With recent advances in X-ray optics9 and the emergence of 4th generation synchrotrons10,11, it becomes conceivable to extend this highly accessible approach to entire brain regions with increasing resolution.
]]></description>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Bosch, C.</dc:creator>
<dc:creator>Ackels, T.</dc:creator>
<dc:creator>Laugros, A.</dc:creator>
<dc:creator>Bonnin, A.</dc:creator>
<dc:creator>Livingstone, J.</dc:creator>
<dc:creator>Waltenberg, C.</dc:creator>
<dc:creator>Berning, M.</dc:creator>
<dc:creator>Tootoonian, S.</dc:creator>
<dc:creator>Kollo, M.</dc:creator>
<dc:creator>Nathansen, A.</dc:creator>
<dc:creator>Rzepka, N.</dc:creator>
<dc:creator>Cloetens, P.</dc:creator>
<dc:creator>Pacureanu, A.</dc:creator>
<dc:creator>Schaefer, A.</dc:creator>
<dc:date>2025-04-27</dc:date>
<dc:identifier>doi:10.1101/2025.04.24.650439</dc:identifier>
<dc:title><![CDATA[Structure-Function Mapping of Olfactory Bulb Circuits with Synchrotron X-ray Nanotomography]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.04.25.650684v1?rss=1">
<title>
<![CDATA[
Chromosome-Specific Aneuploidy Engineering via dCas9-Induced Centromeric Chromatin Relaxation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.04.25.650684v1?rss=1"
</link>
<description><![CDATA[
Aneuploidy, the gain or loss of chromosomes, is prevalent in both normal and disease conditions, however, experimental approaches to engineer and study aneuploidy remain limited, leaving its functional significance under-characterized. Here, we present CRISPR-Taiji (CRISPRt), an efficient method for inducing chromosome-specific mis-segregation and aneuploidy generation across all 24 human chromosomes via dead Cas9 (dCas9)-induced centromeric chromatin relaxation. Using CRISPRt with scRNA-seq, we generated the first comprehensive transcriptomic alteration landscape of nearly all autosomal aneuploidies at chromosome-arm resolution. This genotype-phenotype map provides causal evidence linking recurrent aneuploidies in clear cell renal cell carcinoma (ccRCC) to molecular and clinical phenotypes observed in patient tumors. Notably, chromosome 3(p) loss, the ccRCC initiating event, specifically drives strong interferon signaling activation, offering novel insights into ccRCC tumorigenesis and immune modulation. Overall, we establish CRISPRt as a simple, efficient and scalable approach for chromosome-specific aneuploidy engineering and characterization in preclinical models to advance aneuploidy research across diverse biological contexts.
]]></description>
<dc:creator>FENG, H.</dc:creator>
<dc:creator>Deng, D.</dc:creator>
<dc:creator>Dahiya, R.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Zeng, J.</dc:creator>
<dc:creator>Tan, B. J. Y.</dc:creator>
<dc:creator>Byrne, F.</dc:creator>
<dc:creator>Shepherd, S. T. C.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Johnson, S. C.</dc:creator>
<dc:creator>Harrod, A.</dc:creator>
<dc:creator>Lane, K. A.</dc:creator>
<dc:creator>Fendler, A.</dc:creator>
<dc:creator>Cattin, A.-L.</dc:creator>
<dc:creator>Tippu, Z.</dc:creator>
<dc:creator>Nie, M.</dc:creator>
<dc:creator>Zhao, Y.</dc:creator>
<dc:creator>Wang, R.</dc:creator>
<dc:creator>Ai, W.</dc:creator>
<dc:creator>Bouricha, O.</dc:creator>
<dc:creator>Barber, T.</dc:creator>
<dc:creator>Dovga, Y.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Devito, L. G.</dc:creator>
<dc:creator>Healy, L.</dc:creator>
<dc:creator>Sun, Z.</dc:creator>
<dc:creator>Shen, W.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Lim, E. S.</dc:creator>
<dc:creator>OToole, S. M.</dc:creator>
<dc:creator>Akker, S.</dc:creator>
<dc:creator>Drake, W. M.</dc:creator>
<dc:creator>Jin, H.</dc:creator>
<dc:creator>Downs, J. A.</dc:creator>
<dc:creator>McClelland, S. E.</dc:creator>
<dc:creator>Diffley, J. F. C.</dc:creator>
<dc:creator>Ly, P.</dc:creator>
<dc:creator>Turajlic, S.</dc:creator>
<dc:date>2025-04-27</dc:date>
<dc:identifier>doi:10.1101/2025.04.25.650684</dc:identifier>
<dc:title><![CDATA[Chromosome-Specific Aneuploidy Engineering via dCas9-Induced Centromeric Chromatin Relaxation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.04.25.650693v1?rss=1">
<title>
<![CDATA[
VGLL1 contributes to both the transcriptome and epigenome of the developing trophoblast compartment 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.04.25.650693v1?rss=1"
</link>
<description><![CDATA[
The trophectoderm (TE), the first lineage specified during mammalian development, initiates implantation and gives rise to placental trophoblasts. While animal models have elucidated key conserved signaling pathways involved in early TE specification, including BMP, WNT, and HIPPO, species-specific differences during early development emphasize the need for human-specific models. We previously identified VGLL1, a coactivator of TEAD transcription factors, as a human-specific placental marker. In this study, we employed a pluripotent stem cell (PSC)-based model of TE induction by BMP4 to investigate chromatin remodeling and transcriptional dynamics during TE formation. BMP4-induced chromatin accessibility changes promoted a trophoblast gene expression program, while mesoderm lineage markers were only transiently expressed upon canonical WNT activation. We found that VGLL1 was expressed downstream of key TE transcription factors (GATA2/3, TFAP2A/C) but was essential for establishment of full trophoblast identity by up-regulating EGFR and reinforcing GATA3 expression through positive feedback. Notably, VGLL1 enhanced canonical WNT signaling via direct regulation of WNT receptors and effectors. We also identified KDM6B, a histone demethylase that removes H3K27me3 repressive marks, as a direct VGLL1 target. KDM6B facilitated activation of bivalent promoters associated with TE markers, linking epigenetic regulation to lineage identity. Our findings establish a mechanistic framework positioning VGLL1 as a central regulator that integrates HIPPO, BMP, and WNT signaling pathways to drive establishment of human TE.

Statement of SignificanceEarly development of the human placenta is essential for pregnancy success, yet the mechanisms that guide placental lineage specification remain poorly defined. Using human stem cells, we show how signaling pathways and chromatin remodeling programs work together to direct formation of the trophectoderm, the earliest placental cell type. We identify VGLL1 as a key regulator linking multiple signaling networks to gene expression and epigenetic control. Our findings reveal a species-specific mechanism of placental initiation with broad implications for understanding reproductive disorders, pregnancy loss, and advancing stem cell-based models to study and potentially treat human placental disease.
]]></description>
<dc:creator>Calderon, R. I.</dc:creator>
<dc:creator>Sah, N.</dc:creator>
<dc:creator>Huang, M.</dc:creator>
<dc:creator>Kittle, R. H.</dc:creator>
<dc:creator>Shaik, W.</dc:creator>
<dc:creator>Chousal, J. N.</dc:creator>
<dc:creator>Kallol, S.</dc:creator>
<dc:creator>Bui, T.</dc:creator>
<dc:creator>Morey, R.</dc:creator>
<dc:creator>Mitre, A.</dc:creator>
<dc:creator>Fogarty, N. M. E.</dc:creator>
<dc:creator>Gerri, C.</dc:creator>
<dc:creator>Zheng, C.</dc:creator>
<dc:creator>DeHoff, P. M.</dc:creator>
<dc:creator>Home, P.</dc:creator>
<dc:creator>Niakan, K.</dc:creator>
<dc:creator>Cook-Andersen, H.</dc:creator>
<dc:creator>Fisch, K. M.</dc:creator>
<dc:creator>Paul, S.</dc:creator>
<dc:creator>Soncin, F.</dc:creator>
<dc:date>2025-04-29</dc:date>
<dc:identifier>doi:10.1101/2025.04.25.650693</dc:identifier>
<dc:title><![CDATA[VGLL1 contributes to both the transcriptome and epigenome of the developing trophoblast compartment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.04.28.650978v1?rss=1">
<title>
<![CDATA[
Diverse Genomic Landscape of Swine Influenza A Virus in England (2014 - 2021) 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.04.28.650978v1?rss=1"
</link>
<description><![CDATA[
Surveillance of influenza A viruses in pigs (SwIAV) is critical for identification of novel genetic groups that pose a risk to pig health and might have zoonotic potential. SwIAVs circulating in pigs in England between 2014 and 2021 were characterised using whole genome sequencing (WGS).

Haemagglutinin (HA) and neuraminidase (NA) sequencing data from 82 of 368 influenza A positive samples (71 submissions) were determined, identifying H1N1 and H1N2 subtypes from the 1A classical swine and 1B human-seasonal lineages respectively. The 1B lineage viruses were predominant, accounting for 68.29% of sequenced viruses, with 1A lineage viruses comprising 31.71%, primarily from the 1A.3.3.2 clade (2009 H1N1 pandemic origin). This study characterised previously undefined diversity within the 1B lineage which led to the designation of new HA clades 1B.1.1.1, 1B.1.1.2 and 1B.1.1.3. Complete genome data were obtained from 64/82 viruses thereby updating the definition of genetic diversity thresholds and leading to the identification of 24 unique genotypes. All these 64 viruses contained PB2, PB1, PA, NP, MP, and NS gene segments of 2009 H1N1 pandemic origin. These data highlight the increasing divergence of SwIAV within pig populations England and emphasise the requirement for continued genomic surveillance to improve animal health and monitor zoonotic risk.
]]></description>
<dc:creator>Mollett, B. C.</dc:creator>
<dc:creator>Byrne, A. M. P.</dc:creator>
<dc:creator>Everett, H. E.</dc:creator>
<dc:creator>Reid, S. M.</dc:creator>
<dc:creator>Williamson, S.</dc:creator>
<dc:creator>Anderson, T. K.</dc:creator>
<dc:creator>James, J.</dc:creator>
<dc:creator>Banyard, A. C.</dc:creator>
<dc:creator>Brown, I. H.</dc:creator>
<dc:creator>Lewis, N. S.</dc:creator>
<dc:date>2025-04-30</dc:date>
<dc:identifier>doi:10.1101/2025.04.28.650978</dc:identifier>
<dc:title><![CDATA[Diverse Genomic Landscape of Swine Influenza A Virus in England (2014 - 2021)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.04.25.650707v1?rss=1">
<title>
<![CDATA[
Distinct druggable biological processes in early-onset prostate cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.04.25.650707v1?rss=1"
</link>
<description><![CDATA[
Despite advances in understanding and treating Prostate Cancer (PCa), there has been little effort to systematically map the biology distinguishing Early-(EOPCa) and Late-(LOPCa) onset PCa. Around 25% of EOPCa cases present with metastatic spread or aggressive disease with earlier metastatic development. Some available lines of therapy are extending treatment trajectories and prolonging lives. However, there remains a critical clinical need to identify new therapeutic targets for EOPCa where life expectancy necessitates safer, more targeted treatment options. To our knowledge, here we present the largest systematic analysis of molecular profiles in EOPCa versus LOPCa, employing machine-learning-enabled algorithms to identify distinguishing biology and druggable targets for each age group. Distinct stromal signatures are uncovered in EOPCa, which are used to propose therapeutic opportunities herein. Moreover, our analysis identifies 50 druggable targets, 11 of which we confirm in PCa cell line genetic/pharmacological perturbation data. These findings provide the first specific, testable hypotheses in EOPCa, offering avenues for experimental validation and potential therapeutic exploitation, and, more generally, shed light on the intricate and distinguished molecular profile of this aggressive, poorly understood disease.

One Sentence SummaryMachine learning-enabled algorithms were utilized to identify distinguishing biology and associated druggable targets for early-onset prostate cancers.
]]></description>
<dc:creator>Schmidt, S.</dc:creator>
<dc:creator>Jarai, Z.</dc:creator>
<dc:creator>Larkeryd, A.</dc:creator>
<dc:creator>Ozer, B.</dc:creator>
<dc:creator>Karlsson, Q.</dc:creator>
<dc:creator>Campbell, J.</dc:creator>
<dc:creator>Merson, S.</dc:creator>
<dc:creator>Dennis, N.</dc:creator>
<dc:creator>Hazell, S.</dc:creator>
<dc:creator>Mitsopoulos, C.</dc:creator>
<dc:creator>Cooper, C.</dc:creator>
<dc:creator>ICGC-UK, T.</dc:creator>
<dc:creator>Consortium, T.</dc:creator>
<dc:creator>Russell, K.</dc:creator>
<dc:creator>Workman, P.</dc:creator>
<dc:creator>Eeles, R.</dc:creator>
<dc:creator>Al-Lazikani, B.</dc:creator>
<dc:date>2025-04-29</dc:date>
<dc:identifier>doi:10.1101/2025.04.25.650707</dc:identifier>
<dc:title><![CDATA[Distinct druggable biological processes in early-onset prostate cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.04.30.651378v1?rss=1">
<title>
<![CDATA[
PEtab.jl: Advancing the Efficiency and Utility of Dynamic Modelling 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.04.30.651378v1?rss=1"
</link>
<description><![CDATA[
Dynamic models are useful to study processes ranging from cell signalling to cell differentiation. Common modelling workflows such as model exploration and parameter estimation are computationally demanding. The Julia programming language is a promising tool to address these computational challenges. To evaluate it we developed SBMLImporter.jl and PEtab.jl, a package for model fitting. SBMLImporter.jl was used to evaluate different stochastic simulators against PySB and RoadRunner, overall Julia simulators proved fastest. For Ordinary Differential Equations (ODE) models solvers, gradient methods, and parameter estimation performance were evaluated using PEtab benchmark problems. For the latter two tasks PEtab.jl was compared against pyPESTO, which employs the high-performance AMICI library. Guidelines for choosing ODE solver were produced by evaluating 31 ODE solvers for 29 models. Further, by leveraging automatic differentiation PEtab.jl proved efficient and, for up to medium-sized models, was often at least twice faster than pyPESTO, showcasing how Julias ecosystem can accelerate modelling workflows.
]]></description>
<dc:creator>Cvijovic, M.</dc:creator>
<dc:creator>Persson, S.</dc:creator>
<dc:creator>Frohlich, F.</dc:creator>
<dc:creator>Grein, S.</dc:creator>
<dc:creator>Lomna, T.</dc:creator>
<dc:creator>Ognissanti, D.</dc:creator>
<dc:creator>Hassselgren, V.</dc:creator>
<dc:creator>Hasenauer, J.</dc:creator>
<dc:date>2025-05-04</dc:date>
<dc:identifier>doi:10.1101/2025.04.30.651378</dc:identifier>
<dc:title><![CDATA[PEtab.jl: Advancing the Efficiency and Utility of Dynamic Modelling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.03.652017v1?rss=1">
<title>
<![CDATA[
A single cell ATAC-seq atlas uncovers dynamic changes in chromatin accessibility during cell fate specification at the neural plate border 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.03.652017v1?rss=1"
</link>
<description><![CDATA[
During development, dynamic changes in gene expression and chromatin architecture drive the transition from progenitors to specialised cell types. Here we use single cell ATAC sequencing (scATAC-seq) to investigate changes in chromatin accessibility as neural plate border cells segregate into neural, neural crest and placode cells. We developed a Nextflow pipeline,  single cell Advanced Chromatin Exploration (scACE), which integrates scATAC-seq and scRNA-seq data to identify cell state specific accessibility profiles and groups of chromatin regions with coordinated dynamic behaviour, termed accessibility modules (AMs). We find that progenitors are characterised by broadly open chromatin, reflecting their broad potential to generate any ectodermal derivative. As development proceeds, cell-type-specific chromatin signatures are established. Inferring an enhancer-centric gene regulatory network, we predict Foxk2 as new regulator for placode specification and verify this prediction experimentally. Foxk2 target enhancers are open in placodal, but not any other ectodermal cells. This finding suggests that on a regulatory level, cells can use different strategies to control fate choice: differential accessibility of enhancers and broad accessibility controlled by differentially expressed transcription factors.
]]></description>
<dc:creator>Harmrud, E.</dc:creator>
<dc:creator>Leese, J.</dc:creator>
<dc:creator>Thiery, A. P.</dc:creator>
<dc:creator>Buzzi, A. L.</dc:creator>
<dc:creator>Vigilante, A.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Streit, A.</dc:creator>
<dc:date>2025-05-05</dc:date>
<dc:identifier>doi:10.1101/2025.05.03.652017</dc:identifier>
<dc:title><![CDATA[A single cell ATAC-seq atlas uncovers dynamic changes in chromatin accessibility during cell fate specification at the neural plate border]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.02.651738v1?rss=1">
<title>
<![CDATA[
A spatial code governs olfactory receptor choice and aligns sensory maps in the nose and brain 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.02.651738v1?rss=1"
</link>
<description><![CDATA[
Although topographical maps organize many peripheral sensory systems, it remains unclear whether olfactory sensory neurons (OSNs) choose which of the [~]1100 odor receptors (ORs) to express based upon their spatial location in the olfactory epithelium (OE) or instead ORs are scattered randomly. Here we reveal that each OR is expressed at a precise mean position along the OE dorsoventral axis, thereby instantiating a receptor map. This patterning reflects the differential use, by precursors and mature OSNs, of a coherent gene expression program controlled by a spatially-varying retinoic acid gradient; this program -- which includes key transcription factors and axon guidance genes -- translates position into a spatially appropriate distribution of OR choices and aligns the epithelial map of OR identity with the glomerular map present in the olfactory bulb. These results identify a transcriptional code that distinguishes and spatially organizes the vast array of sensory channels that comprise the olfactory system.
]]></description>
<dc:creator>Brann, D. H.</dc:creator>
<dc:creator>Tsukahara, T.</dc:creator>
<dc:creator>Tau, C.</dc:creator>
<dc:creator>Kalloor, D.</dc:creator>
<dc:creator>Lubash, R.</dc:creator>
<dc:creator>Thamarai Kannan, L.</dc:creator>
<dc:creator>Klimpert, N.</dc:creator>
<dc:creator>Kollo, M.</dc:creator>
<dc:creator>Escamilla-Del-Arenal, M.</dc:creator>
<dc:creator>Bintu, B.</dc:creator>
<dc:creator>Bozza, T.</dc:creator>
<dc:creator>Datta, S. R.</dc:creator>
<dc:date>2025-05-08</dc:date>
<dc:identifier>doi:10.1101/2025.05.02.651738</dc:identifier>
<dc:title><![CDATA[A spatial code governs olfactory receptor choice and aligns sensory maps in the nose and brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.05.652027v1?rss=1">
<title>
<![CDATA[
Sex-dependent neural plasticity in response to damage 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.05.652027v1?rss=1"
</link>
<description><![CDATA[
Plasticity of intact adult neural tissue in the vicinity of neural damage serves to restore functionality of circuits. Much remains to be learned about the mechanisms regulating this process and the reported sex differences in recovery outcomes. Here, we present the fly gut and its innervation as a simplified model to address these questions. We show that gut damage caused by ingestion of toxic agents resulted in plasticity of the adult enteric neuronal network, manifested as a Reactive Oxygen Species (ROS)-dependent increase in neural tissue, which was reversible after a recovery period. Interestingly, males did not display neural plasticity, and masculinization of neurons in females suppressed the damage-dependent neural growth. Together, these findings position the fly gut as a system to investigate the cellular, molecular, and sex-specific underpinnings of neural plasticity, with implications for therapeutic advancements in neural circuit recovery.

SUMMARY STATEMENTThis study establishes the fly gut as a simple system to explore how adult neural tissues differ between sexes in their capacity for plasticity.
]]></description>
<dc:creator>Recatala-Martinez, M.</dc:creator>
<dc:creator>Bosch, M.</dc:creator>
<dc:creator>Gaspar, P.</dc:creator>
<dc:creator>Mineo, A.</dc:creator>
<dc:creator>Rios, S.</dc:creator>
<dc:creator>Miguel-Aliaga, I.</dc:creator>
<dc:creator>Morey, M.</dc:creator>
<dc:date>2025-05-09</dc:date>
<dc:identifier>doi:10.1101/2025.05.05.652027</dc:identifier>
<dc:title><![CDATA[Sex-dependent neural plasticity in response to damage]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.12.653400v1?rss=1">
<title>
<![CDATA[
Mycobacterium tuberculosis partitions the Krebs cycle to persist under iron starvation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.12.653400v1?rss=1"
</link>
<description><![CDATA[
In this study, we investigated how iron limitation alters central metabolism in Mycobacterium tuberculosis using metabolomics and stable isotope tracing. Our findings reveal a well-orchestrated metabolic program to enable Krebs cycle activity despite the inefficient action of its iron-dependent enzymes. Under such conditions, carbon flux through the oxidative branch of the Krebs cycle is stalled, resulting in the accumulation of metabolites that are partially secreted. As a result, carbon flux from glycolysis is partially diverted to the reductive branch of the Krebs cycle to support the production of oxaloacetate and malate through the activity of phosphoenolpyruvate carboxykinase and pyruvate carboxylase. Both branches terminate with the synthesis of malate, which is secreted. This unprecedented split of the Krebs cycle and malate secretion in a bacterial pathogen facilitates the continuous flow of carbon through the core of carbon metabolism, overcoming the metabolic stalling triggered by iron starvation.
]]></description>
<dc:creator>Serafini, A.</dc:creator>
<dc:creator>Garza-Garcia, A.</dc:creator>
<dc:creator>Sorze, D.</dc:creator>
<dc:creator>de Carvalho, L. P. S.</dc:creator>
<dc:creator>Manganelli, R.</dc:creator>
<dc:date>2025-05-12</dc:date>
<dc:identifier>doi:10.1101/2025.05.12.653400</dc:identifier>
<dc:title><![CDATA[Mycobacterium tuberculosis partitions the Krebs cycle to persist under iron starvation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.12.653404v1?rss=1">
<title>
<![CDATA[
Conformational dynamics of the bacterial E3 ligase SspH1 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.12.653404v1?rss=1"
</link>
<description><![CDATA[
The SspH/IpaH family of novel E3 ligases (NELs) are found in a number of Gram-negative bacteria and are used to target host enzymes for degradation to support pathogenesis. These E3 enzymes are autoinhibited in the absence of substrate and different models for release of autoinhibition have been suggested. However, many of the molecular details of individual steps during the ubiquitin transfer reaction remain unknown. Here, we present the crystal structure of Salmonella SspH1 and an analysis of the solution properties of SspH1 on its own and in complex with substrate and ubiquitin. Our data show that SspH1 exists in a conformational equilibrium between open and closed states and that substrate binding only modulates the distribution of these states but does not induce major conformational changes. This suggests that additional mechanisms must exist to bring the substrates close to the active site to mediate transfer of ubiquitin from the E3[~]Ub conjugate.
]]></description>
<dc:creator>Kennedy, C. R.</dc:creator>
<dc:creator>Esposito, D.</dc:creator>
<dc:creator>House, D.</dc:creator>
<dc:creator>Rittinger, K.</dc:creator>
<dc:date>2025-05-15</dc:date>
<dc:identifier>doi:10.1101/2025.05.12.653404</dc:identifier>
<dc:title><![CDATA[Conformational dynamics of the bacterial E3 ligase SspH1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.12.653464v1?rss=1">
<title>
<![CDATA[
AI4CellFate: Interpretable Early Cell Fate Prediction with Generative AI 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.12.653464v1?rss=1"
</link>
<description><![CDATA[
Live-cell imaging provides a unique insight into complex cellular processes including single cell fate, but remains limited by both low-throughput and the lack of generalisable analytics for the multidimensional datasets it produces. This work introduces AI4CellFate, an interpretable and data-driven machine learning framework for predicting cell fate from microscopy timelapses, applied here to cancer therapy. By integrating generative AI and contrastive learning, AI4CellFate enables early fate prediction as well as visualisation of biologically relevant features, with limited annotation.
]]></description>
<dc:creator>Cunha, I.</dc:creator>
<dc:creator>Panconi, L.</dc:creator>
<dc:creator>Bauer, S.</dc:creator>
<dc:creator>Gestin, M.</dc:creator>
<dc:creator>Latron, E.</dc:creator>
<dc:creator>Sahai, E.</dc:creator>
<dc:creator>Le Marois, A.</dc:creator>
<dc:creator>Griffie, J.</dc:creator>
<dc:date>2025-05-14</dc:date>
<dc:identifier>doi:10.1101/2025.05.12.653464</dc:identifier>
<dc:title><![CDATA[AI4CellFate: Interpretable Early Cell Fate Prediction with Generative AI]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.14.653932v1?rss=1">
<title>
<![CDATA[
Physiological febrile heat stress increases cytoadhesion through increased protein trafficking of Plasmodium falciparum surface proteins into the red blood cell 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.14.653932v1?rss=1"
</link>
<description><![CDATA[
Fever is a hallmark of malaria. Several studies have linked febrile temperatures to reduced parasite viability, but also to increased cytoadhesion, a key driver of pathology. However, different mechanisms have been proposed to cause changes in cytoadhesion and parasite sensitivity to heat. Here, we demonstrate that exposure of Plasmodium falciparum-infected red blood cells (iRBCs) to physiologically relevant febrile heat stress (39 {degrees}C), derived from patient data, enhances cytoadhesion through increased trafficking of the major virulence factor PfEMP1 to the iRBC surface. This phenomenon is not limited to PfEMP1 and common laboratory strains, as it extends to the surface nutrient channel PSAC in four clinical isolates of diverse geographic origin. The increased surface protein display occurs without changes in overall protein expression or parasite developmental progression. Using phosphoproteomics and proximity labelling, we find that elevated temperature also increases trafficking and phosphorylation of exported proteins into the RBC. Enhanced export is likely reliant on the presence of a transmembrane domain as shown by NanoLuc reporter assays. Collectively, our results indicate that febrile temperatures commonly experienced during infection can accelerate protein export, likely at the parasitophorous vacuole. This enhanced export following heat stress is relevant because increased cytoadhesion could influence disease severity through earlier iRBC sequestration and elevated bound parasite mass.
]]></description>
<dc:creator>Jones, D.</dc:creator>
<dc:creator>Belda, H.</dc:creator>
<dc:creator>Fuchs, G.</dc:creator>
<dc:creator>Anaguano, D.</dc:creator>
<dc:creator>Nofal, S. D.</dc:creator>
<dc:creator>Treeck, M.</dc:creator>
<dc:date>2025-05-14</dc:date>
<dc:identifier>doi:10.1101/2025.05.14.653932</dc:identifier>
<dc:title><![CDATA[Physiological febrile heat stress increases cytoadhesion through increased protein trafficking of Plasmodium falciparum surface proteins into the red blood cell]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.13.653495v1?rss=1">
<title>
<![CDATA[
A spatiotemporal cancer cell trajectory underlies glioblastoma heterogeneity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.13.653495v1?rss=1"
</link>
<description><![CDATA[
Cancer cells display highly heterogeneous and plastic states in glioblastoma, an incurable brain tumour. However, how these malignant states arise and whether they follow defined cellular trajectories across tumours is poorly understood. Here, we generated a deep single cell and spatial multi-omic atlas of human glioblastoma that pairs transcriptomic, epigenomic and genomic profiling of 12 tumours across multiple regions. We identify that glioblastoma heterogeneity is driven by spatially-patterned transitions of cancer cells from developmental-like states towards those defined by a glial injury response and hypoxia. This cellular trajectory regionalises tumours into distinct tissue niches and manifests in a molecularly conserved manner across tumours as well as genetically distinct tumour subclones. Moreover, using a new deep learning framework to map cancer cell states jointly with clones in situ, we show that tumour subclones are finely spatially intermixed through glioblastoma tissue niches. Finally, we show that this cancer cell trajectory is intimately linked to myeloid heterogeneity and unfolds across regionalised myeloid signalling environments. Our findings define a stereotyped trajectory of cancer cells in glioblastoma and unify glioblastoma tumour heterogeneity into a tractable cellular and tissue framework.
]]></description>
<dc:creator>de Jong, G.</dc:creator>
<dc:creator>Memi, F.</dc:creator>
<dc:creator>Gracia, T.</dc:creator>
<dc:creator>Lazareva, O.</dc:creator>
<dc:creator>Gould, O.</dc:creator>
<dc:creator>Aivazidis, A.</dc:creator>
<dc:creator>Dave, M.</dc:creator>
<dc:creator>Zhang, Q.</dc:creator>
<dc:creator>Jensen, M.</dc:creator>
<dc:creator>Rifaioglu, A. S.</dc:creator>
<dc:creator>Barros-Silva, J. D.</dc:creator>
<dc:creator>Eckert, S.</dc:creator>
<dc:creator>Zhou, D.</dc:creator>
<dc:creator>Wood, Y.</dc:creator>
<dc:creator>Tuck, E.</dc:creator>
<dc:creator>Er, S.</dc:creator>
<dc:creator>Marshall, H.</dc:creator>
<dc:creator>Roberts, K.</dc:creator>
<dc:creator>Trinh, A. L.</dc:creator>
<dc:creator>Rai, S.</dc:creator>
<dc:creator>Shaw, T.</dc:creator>
<dc:creator>Oszlanczi, A.</dc:creator>
<dc:creator>Powell, H.</dc:creator>
<dc:creator>Petryszak, R.</dc:creator>
<dc:creator>Katsirea, Z.</dc:creator>
<dc:creator>Mamun, I.</dc:creator>
<dc:creator>Mulas, I.</dc:creator>
<dc:creator>Quaegebeur, A.</dc:creator>
<dc:creator>Briggs, M.</dc:creator>
<dc:creator>Makarchuk, S.</dc:creator>
<dc:creator>Cox, J.</dc:creator>
<dc:creator>Lee, J. T. H.</dc:creator>
<dc:creator>Rueda, L.</dc:creator>
<dc:creator>Saraswat, M.</dc:creator>
<dc:creator>Bulstrode, H.</dc:creator>
<dc:creator>Young, A.</dc:creator>
<dc:creator>Patel, M.</dc:creator>
<dc:creator>Porter, T.</dc:creator>
<dc:creator>Prigmore, E.</dc:creator>
<dc:creator>Mall, M.</dc:creator>
<dc:creator>Saez-Rodriguez, J.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Rowitch, D. H.</dc:creator>
<dc:creator>Mair, R.</dc:creator>
<dc:creator>Behjati, S.</dc:creator>
<dc:creator>Stegle, O.</dc:creator>
<dc:creator>Bayraktar, O. A.</dc:creator>
<dc:date>2025-05-14</dc:date>
<dc:identifier>doi:10.1101/2025.05.13.653495</dc:identifier>
<dc:title><![CDATA[A spatiotemporal cancer cell trajectory underlies glioblastoma heterogeneity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.14.653752v1?rss=1">
<title>
<![CDATA[
Mice discriminate odour source distance via sub-sniff temporal features of odour plumes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.14.653752v1?rss=1"
</link>
<description><![CDATA[
Rodents rely on olfaction to navigate complex environments, particularly where visual cues are limited. Yet how they estimate the distance to an odour source remains unclear. The spatiotemporal dynamics of natural odour plumes, shaped by airflow turbulence, offer valuable cues for locating odour sources. Here, we show that mice can discriminate odour sources placed at different distances by extracting information from the sub-sniff temporal structure of naturalistic odour plumes. Using a wind tunnel and an olfactory virtual reality system, we generated dynamic plumes and demonstrated, through high-throughput automated behaviour, that mice distinguish near from far sources based on odour fluctuations operating faster than their respiratory cycle. Two-photon calcium imaging of olfactory bulb projection neurons revealed that distance-dependent responses are present in a small subset of mitral and tufted cells, and that population activity encodes source distance. Critically, neural responses correlated more strongly with high-frequency plume features than with mean odour concentration. Our results identify a neural basis for distance estimation from odour dynamics and highlight the importance of rapid temporal processing in mammalian olfaction.
]]></description>
<dc:creator>Marin, A. C.</dc:creator>
<dc:creator>Harris, J. J.</dc:creator>
<dc:creator>Dasgupta, D.</dc:creator>
<dc:creator>Erskine, A.</dc:creator>
<dc:creator>Warner, T. P. A.</dc:creator>
<dc:creator>Schaefer, A. T.</dc:creator>
<dc:creator>Ackels, T.</dc:creator>
<dc:date>2025-05-17</dc:date>
<dc:identifier>doi:10.1101/2025.05.14.653752</dc:identifier>
<dc:title><![CDATA[Mice discriminate odour source distance via sub-sniff temporal features of odour plumes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.28.656648v1?rss=1">
<title>
<![CDATA[
Dynamic Landscape Analysis of Cell Fate Decisions: Predictive Models of Neural Development From Single-Cell Data 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.28.656648v1?rss=1"
</link>
<description><![CDATA[
Building a mechanistic understanding of cell fate decisions remains a fundamental goal of developmental biology, with implications for stem cell therapies, regenerative medicine and understanding disease mechanisms. Single-cell transcriptomics provides a detailed picture of the cellular states observed during these decisions, but building dynamic and predictive models from these data remains a challenge. Here, we present dynamic landscape analysis (DLA), a framework that applies dynamical systems theory to identify stable cell states, map transition pathways, and generate a predictive cell fate decision landscape from single-cell data. Applying this framework to vertebrate neural tube development, revealed that progenitor specification by Sonic Hedgehog (Shh) can be captured in a landscape with an unexpected topology in which initially divergent lineages converge to the same fate through multiple distinct routes. The model accurately predicted cellular responses and cell fate allocation for unseen dynamic signalling regimes. Cross-species validation using human embryonic organoid data demonstrated conservation of this decision-making architecture. By modelling the dynamic responses that drive cell fate decisions, the DLA framework provides a quantitative and generative framework for extracting mechanistic insights from high-dimensional single-cell data.
]]></description>
<dc:creator>Fontaine, M.</dc:creator>
<dc:creator>Vives, M. J. D.</dc:creator>
<dc:creator>Saez, M.</dc:creator>
<dc:creator>Maizels, R.</dc:creator>
<dc:creator>Finnie, E.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Rand, D. A.</dc:creator>
<dc:date>2025-05-28</dc:date>
<dc:identifier>doi:10.1101/2025.05.28.656648</dc:identifier>
<dc:title><![CDATA[Dynamic Landscape Analysis of Cell Fate Decisions: Predictive Models of Neural Development From Single-Cell Data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.30.657122v1?rss=1">
<title>
<![CDATA[
Targeting Neutrophil Extracellular Traps to inhibit Colon Cancer Tumor Necrosis and Metastasis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.30.657122v1?rss=1"
</link>
<description><![CDATA[
Necrosis, conventionally thought of as a passive consequence of aggressive tumor growth, is associated with poor prognosis in colorectal cancer (CRC). We recently discovered that necrosis can be caused by neutrophils and neutrophil extracellular traps (NETs) aggregates driving vascular occlusion within the tumor vasculature in models of breast cancer. Here, we evaluated the role of NETs in inducing necrosis and metastasis in CRC. We found that the numbers of neutrophils primed to form NETs were elevated in the circulation of patients with CRC as compared to controls. CD177Low neutrophils were also elevated, and they showed reduced extravasation capacity with intact ability to form NETs. The extent of necrosis correlated with metastasis (stage IV disease), independent of tumor size, in our human cohort. In both human and murine CRC tumors, necrotic regions were characterized by neutrophil infiltration and NET accumulation, and NET aggregates were observed in the vasculature next to the necrotic regions. Single cell RNA sequencing and spatial transcriptomic analysis of human CRC and liver metastases revealed that necrotic tumors activate pathways associated with increased metastatic potential, including epithelial-to-mesenchymal-transition. Using a mouse model of DNA mismatch repair proficient CRC, we found neutrophil infiltration and NETs increased with tumor progression. Genetic or pharmacological inhibition of NET formation decreased necrosis and metastasis, and importantly enhanced chemotherapy efficacy. Altogether, our findings show that NET formation in human CRC is a key feature of tumor necrosis, that it is associated with metastasis, and further suggest that preventing NET formation may offer clinical benefits to CRC patients.
]]></description>
<dc:creator>Gazzara, E.</dc:creator>
<dc:creator>Adrover, J. M.</dc:creator>
<dc:creator>Lui, A.</dc:creator>
<dc:creator>Han, S.</dc:creator>
<dc:creator>Aminzada, Z.</dc:creator>
<dc:creator>Bhandari, N.</dc:creator>
<dc:creator>Sivetz, N.</dc:creator>
<dc:creator>Shirue, V. S.</dc:creator>
<dc:creator>Shergill, B. S.</dc:creator>
<dc:creator>Curtis, M. B.</dc:creator>
<dc:creator>George, S. C.</dc:creator>
<dc:creator>Cicala, A.</dc:creator>
<dc:creator>Rishi, A.</dc:creator>
<dc:creator>Chung, C.</dc:creator>
<dc:creator>Devoe, C.</dc:creator>
<dc:creator>Huang, H.</dc:creator>
<dc:creator>Weiss, M.</dc:creator>
<dc:creator>Lou, E.</dc:creator>
<dc:creator>Tuveson, D. A.</dc:creator>
<dc:creator>Beyaz, S.</dc:creator>
<dc:creator>Westcott, P. M. K.</dc:creator>
<dc:creator>Egeblad, M.</dc:creator>
<dc:creator>Gholami, S.</dc:creator>
<dc:date>2025-06-03</dc:date>
<dc:identifier>doi:10.1101/2025.05.30.657122</dc:identifier>
<dc:title><![CDATA[Targeting Neutrophil Extracellular Traps to inhibit Colon Cancer Tumor Necrosis and Metastasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.30.656994v1?rss=1">
<title>
<![CDATA[
C9orf72 polyGA knock-in mice exhibit mild motor and proteomic changes consistent with ALS/FTD 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.30.656994v1?rss=1"
</link>
<description><![CDATA[
A GGGGCC repeat expansion in C9orf72 is the most common genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). The repeat expansion is translated into five different dipeptide repeat proteins: polyGA, polyGP, polyGR, polyAP and polyPR. To investigate the effect of polyGA, which is the most abundant dipeptide repeat protein in patient brains, we used CRISPR/Cas9 to insert 400 codon-optimized polyGA repeats immediately downstream of the mouse C9orf72 start codon. This generated (GA)400 knock-in mice driven by the endogenous mouse C9orf72 promoter, coupled with heterozygous C9orf72 reduction. (GA)400 mice develop subtle pathology including mild motor dysfunction characterized by impaired rotarod performance. Quantitative proteomics revealed polyGA expression caused protein alterations in the spinal cord, including changes in previously identified polyGA interactors. Our findings show that (GA)400 mice are a complementary in vivo model to better understand C9ALS/FTD pathology and determine the specific role of single DPRs in disease.
]]></description>
<dc:creator>Milioto, C.</dc:creator>
<dc:creator>Carcole, M.</dc:creator>
<dc:creator>Zanovello, M.</dc:creator>
<dc:creator>Ahmed, M.</dc:creator>
<dc:creator>Nirujogi, R. S.</dc:creator>
<dc:creator>Biggs, D.</dc:creator>
<dc:creator>Katona, E.</dc:creator>
<dc:creator>Glaria, I.</dc:creator>
<dc:creator>Santos, A.</dc:creator>
<dc:creator>Devoy, A.</dc:creator>
<dc:creator>Fratta, P.</dc:creator>
<dc:creator>Alessi, D. R.</dc:creator>
<dc:creator>Davies, B.</dc:creator>
<dc:creator>Greensmith, L.</dc:creator>
<dc:creator>Fisher, E.</dc:creator>
<dc:creator>Isaacs, A. M.</dc:creator>
<dc:date>2025-06-02</dc:date>
<dc:identifier>doi:10.1101/2025.05.30.656994</dc:identifier>
<dc:title><![CDATA[C9orf72 polyGA knock-in mice exhibit mild motor and proteomic changes consistent with ALS/FTD]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.27.656416v1?rss=1">
<title>
<![CDATA[
Lysosomal stress induces amyloid-β aggregate release and reactive transformation in human astrocytes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.27.656416v1?rss=1"
</link>
<description><![CDATA[
Astrocytes are essential for brain homeostasis and are involved in amyloid-{beta} (A{beta}) clearance, but whether they can produce and release A{beta} aggregates remains unclear. Using human iPSC-derived astrocytes, we show that astrocytes cell autonomously generate small, diffusible A{beta} aggregates under baseline conditions. By combining ultrasensitive single-molecule imaging (DNA-PAINT) and immunoassays, we detect intracellular aggregates and their release into the media. Notably, lysosomal membrane damage induced by L-leucyl-L-leucine methyl ester (LLOMe) significantly increases A{beta} aggregate secretion without altering their size or morphology. Transcriptomic analysis and cytokine profiling reveal that lysosomal stress triggers a reactive astrocyte phenotype marked by upregulation of inflammatory genes and secreted cytokines. These findings suggest that astrocytes are not merely passive A{beta} scavengers but can actively contribute to extracellular A{beta} accumulation under lysosomal stress. Our study highlights astrocytes as active players in Alzheimers disease pathology.
]]></description>
<dc:creator>Goldsworthy, S.</dc:creator>
<dc:creator>Fertan, E.</dc:creator>
<dc:creator>Dyal, P.</dc:creator>
<dc:creator>Domoney, S.</dc:creator>
<dc:creator>Livesey, F.</dc:creator>
<dc:creator>Klenerman, D.</dc:creator>
<dc:creator>Patani, R.</dc:creator>
<dc:creator>Hung, C.</dc:creator>
<dc:date>2025-06-01</dc:date>
<dc:identifier>doi:10.1101/2025.05.27.656416</dc:identifier>
<dc:title><![CDATA[Lysosomal stress induces amyloid-β aggregate release and reactive transformation in human astrocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.28.656570v1?rss=1">
<title>
<![CDATA[
Beneficial and detrimental consequences of AHR activation in intestinal infection 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.28.656570v1?rss=1"
</link>
<description><![CDATA[
The ligand dependent transcription factor aryl hydrocarbon receptor (AHR) is an environmental sensor whose activation can have physiologically beneficial or detrimental consequences for host immune responses depending on the ligand. Here we investigated the hypothesis that prolonged AHR activation either due to inefficient ligand metabolism or due to genetic manipulation may underlie the distinction between beneficial and detrimental effects. Our data indicate that prolonged AHR activation caused toxic endpoints for liver and thymus but was not per se interfering with the host response to infection with the intestinal pathogen C.rodentium. Genetically driven constitutive AHR activation improved resistance to infection, whereas prolonged AHR activation by the pollutant TCDD resulted in delayed clearance of C.rodentium associated with a suppression in antibody production. Combined single cell RNAseq and ATAC-seq analysis provided evidence that TCDD, but not genetic AHR activation, negatively affected dendritic cell functions such as activation, maturation and antigen presentation. Thus, the detrimental impact of environmental pollutants such as TCDD on immune responses cannot solely be attributed to aberrantly prolonged activation of AHR.
]]></description>
<dc:creator>Diaz, O. P.</dc:creator>
<dc:creator>zhou, L.</dc:creator>
<dc:creator>Barrington, C.</dc:creator>
<dc:creator>Lindquist, D.</dc:creator>
<dc:creator>Graelmann, F.</dc:creator>
<dc:creator>Wincent, E.</dc:creator>
<dc:creator>Stockinger, B.</dc:creator>
<dc:date>2025-06-01</dc:date>
<dc:identifier>doi:10.1101/2025.05.28.656570</dc:identifier>
<dc:title><![CDATA[Beneficial and detrimental consequences of AHR activation in intestinal infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.27.656346v1?rss=1">
<title>
<![CDATA[
The ribosome synchronizes folding and assembly to promote oligomeric protein biogenesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.27.656346v1?rss=1"
</link>
<description><![CDATA[
Natural proteins are structurally diverse and often form intricate multidomain, oligomeric architectures. This presents a prima facie challenge to cellular homeostasis, as topologically complex proteins seldom refold efficiently in vitro. How cells overcome sequence-intrinsic folding limitations to optimize protein biogenesis is incompletely understood. Here, we show that efficient folding and assembly of the model five-domain homotetramer {beta}-galactosidase is obligatorily coupled to its synthesis on the ribosome, and define the underlying mechanisms. During refolding of full-length protein from denaturant, maturation of the catalytic domain is frustrated. Assembly outpaces monomer folding, and non-native oligomers accumulate. The ribosome directs the order of folding events and specifies the pathway of oligomer assembly. Efficient de novo folding is characterised by segmental domain folding, shaped by binding of a nascent amphipathic helix to a cryptic pocket on the ribosome surface. Homomer assembly initiates cotranslationally via recruitment of a full-length subunit to the nascent polypeptide, and the failure to do so results in misassembly. Our results reveal how the ribosome can dictate the timing of folding and assembly to enable efficient biogenesis of a topologically complex protein.
]]></description>
<dc:creator>Roeselova, A.</dc:creator>
<dc:creator>Shivakumaraswamy, S.</dc:creator>
<dc:creator>Jurkeviciute, G.</dc:creator>
<dc:creator>He, J.-Z.</dc:creator>
<dc:creator>Auburger, J.</dc:creator>
<dc:creator>Schmitt, J.</dc:creator>
<dc:creator>Kramer, G.</dc:creator>
<dc:creator>Bukau, B.</dc:creator>
<dc:creator>Enchev, R. I.</dc:creator>
<dc:creator>Balchin, D.</dc:creator>
<dc:date>2025-05-30</dc:date>
<dc:identifier>doi:10.1101/2025.05.27.656346</dc:identifier>
<dc:title><![CDATA[The ribosome synchronizes folding and assembly to promote oligomeric protein biogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.28.656646v1?rss=1">
<title>
<![CDATA[
The penetrant chordoid glioma PRKCA mutation is an oncogenic gain-of-function kinase inactivation eliciting early onset chondrosarcoma in mice. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.28.656646v1?rss=1"
</link>
<description><![CDATA[
The penetrant PRKCA D463H mutation, a biomarker and potential driver in chordoid glioma, was found to provoke the development of chondrosarcomas in heterozygous knock-in mice. This mutation entirely abrogates kinase activity, but strikingly no oncogenic phenotype is observed for the related inactivating mutation D463N indicating that the lack of activity is not the driver. In cells, the D463H mutant closely mirrored PKC WT behaviours and retained ATP binding, contrary to the related D463N mutant. Mechanistically, the PKC D463H mutant protein was found to display quantitative alterations to the PKC interactome, enhancing association with epigenetic regulators. This aligned with transcriptomic changes which resembled an augmented PKC expression program, with enhanced BRD4, Myc and TGF{beta} signatures. D463H dependent reduced sensitivity to the BET inhibitors JQ1 and AZD5153 indicates the functional importance of these pathways. The data show that D463H is a dominant gain-of-function oncogenic mutant, operating through a non-catalytic allosteric mechanism.

One Sentence SummaryA PKC catalytic inactivating mutation confers gain-of-function properties - a paradigm shift in kinase actions.
]]></description>
<dc:creator>Calleja, v.</dc:creator>
<dc:creator>Henry, J. C.</dc:creator>
<dc:creator>Cobbaut, M.</dc:creator>
<dc:creator>Sewell, J.</dc:creator>
<dc:creator>Rizzoti, K.</dc:creator>
<dc:creator>Houghton, F.</dc:creator>
<dc:creator>Boeing, S.</dc:creator>
<dc:creator>Anyanwu, N.</dc:creator>
<dc:creator>Varsani-Brown, S.</dc:creator>
<dc:creator>Snoeks, T.</dc:creator>
<dc:creator>Suarez-Bonnet, A.</dc:creator>
<dc:creator>Priestnall, S. L.</dc:creator>
<dc:creator>McDonald, N. Q.</dc:creator>
<dc:creator>Cameron, A. J.</dc:creator>
<dc:creator>Parker, P. J.</dc:creator>
<dc:date>2025-05-29</dc:date>
<dc:identifier>doi:10.1101/2025.05.28.656646</dc:identifier>
<dc:title><![CDATA[The penetrant chordoid glioma PRKCA mutation is an oncogenic gain-of-function kinase inactivation eliciting early onset chondrosarcoma in mice.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.27.655523v1?rss=1">
<title>
<![CDATA[
MitoTracker transfers from astrocytes to neurons independently of mitochondria 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.27.655523v1?rss=1"
</link>
<description><![CDATA[
The neuroprotective transfer of mitochondria from astrocytes to neurons has been primarily investigated by labelling astrocytic mitochondria with the dye MitoTracker. Here we report that MitoTracker transfers to neurons from both astrocytes and astrocyte-conditioned media, independently of mitochondrial transfer. Our observations should prompt an essential re-evaluation of the literature concerning astrocyte-neuron mitochondrial transfer and in other systems in which contact-independent transfer has been observed using mitochondrial dyes.
]]></description>
<dc:creator>Hole, K. L.</dc:creator>
<dc:creator>Norkett, R.</dc:creator>
<dc:creator>Russell, E.</dc:creator>
<dc:creator>Strom, M.</dc:creator>
<dc:creator>Howden, J. H.</dc:creator>
<dc:creator>Corbett, N. J.</dc:creator>
<dc:creator>Brownlees, J.</dc:creator>
<dc:creator>Devine, M. J.</dc:creator>
<dc:date>2025-05-28</dc:date>
<dc:identifier>doi:10.1101/2025.05.27.655523</dc:identifier>
<dc:title><![CDATA[MitoTracker transfers from astrocytes to neurons independently of mitochondria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.24.655746v1?rss=1">
<title>
<![CDATA[
RAD51AP1 is a versatile RAD51 modulator 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.24.655746v1?rss=1"
</link>
<description><![CDATA[
RAD51AP1 is an emergent key factor in homologous recombination (HR), the major pathway for accurate repair of DNA double-strand breaks, and in alternative lengthening of telomeres (ALT). Depletion of RAD51AP1 diminishes HR and overexpression is common in cancer, where it is associated with malignancy. Here, we show that RAD51AP1 serves as a versatile modulator of the RAD51 recombinase, the central player in HR. Through a combination of biochemistry and structural biology, we reveal that RAD51AP1 possesses at least three RAD51-binding sites that facilitate its binding across two adjacent RAD51 molecules. We uncover a novel RAD51 binding mode and characterise a hitherto overlooked role for RAD51AP1 in stabilising RAD51-ssDNA filaments and promoting strand exchange. Further, we resolved structures of RAD51-ssDNA filaments in the presence of Mg2+-ATP and Mg2+-ADP, revealing conformational changes upon ATP hydrolysis and explaining how ADP reduces RAD51-DNA binding. Our findings provide mechanistic insights into RAD51 recombinase and RAD51AP1.

HighlightsO_LIStructures of RAD51 filaments in the presence of Mg2+-ATP and Mg2+-ADP reveal how ATP hydrolysis reduces DNA binding
C_LIO_LIATP hydrolysis does not induce RAD51 filament contraction, instead it induces filament relaxation
C_LIO_LIRAD51AP1 uses three sites to bind across two RAD51 monomers
C_LIO_LIRAD51AP1 uses a unique binding mode to stabilise the RAD51 N-terminal domain and protomer interface of RAD51
C_LIO_LIRAD51AP1 binding induces conformational changes that promote RAD51 DNA association, oligomerisation, filament nucleation, filament stabilisation and strand exchange
C_LI
]]></description>
<dc:creator>Kuhlen, L.</dc:creator>
<dc:creator>Argunhan, B.</dc:creator>
<dc:creator>Liang, P.</dc:creator>
<dc:creator>Zhong, J.</dc:creator>
<dc:creator>Masino, L.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:date>2025-05-24</dc:date>
<dc:identifier>doi:10.1101/2025.05.24.655746</dc:identifier>
<dc:title><![CDATA[RAD51AP1 is a versatile RAD51 modulator]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.19.653976v1?rss=1">
<title>
<![CDATA[
Protein Kinase C δ: a critical hub regulating macrophage immunomodulatory functions during Mycobacterium tuberculosis infection 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.19.653976v1?rss=1"
</link>
<description><![CDATA[
A host-modulating candidate gene involved in putative pathogen-killing pathways, with potential novel therapeutic intervention, Protein Kinase C - {delta} (PKC{delta}) has been recognized as a critical marker of inflammation with clinical and experimental evidence in recent years. Pulmonary microenvironment during Mtb infection is largely governed by lung resident macrophages, initiating innate and subsequent adaptive immune responses. We investigated the role of PKC{delta} in macrophages using a macrophage-specific PKC{delta} knockout mice model (LysMcrePKC{delta}flox/flox). PKC{delta} deficiency in macrophages triggers an early lymphocytic immune response, increases neutrophil recruitment, and reduces inflammatory macrophages in the lungs, leading to higher Mtb burden and exacerbated pathology. Experimental and omics analysis further revealed that dysregulation of antimicrobial effector functions is detrimental to macrophages ability to restrict bacterial growth in vitro. Importantly this defect was mitigated by exogenous GM-CSF supplementation and/or overexpressing PKC{delta} in macrophages. Thus, PKC{delta} plays a crucial role in immune modulation during Mtb infection with GM-CSF amongst several downstream pathways through which PKC{delta} exerts its regulatory effects.

TeaserPKC{delta} is crucial for immune modulation during Mtb infection revealing macrophages as a potential axis of signaling.
]]></description>
<dc:creator>Hazra, R.</dc:creator>
<dc:creator>Ozturk, M.</dc:creator>
<dc:creator>Peton, N.</dc:creator>
<dc:creator>Ganief, T.</dc:creator>
<dc:creator>Poswayo, S. K.</dc:creator>
<dc:creator>Rousseau, R. P.</dc:creator>
<dc:creator>Naidoo, S.</dc:creator>
<dc:creator>Jones, S.-S.</dc:creator>
<dc:creator>Savulescu, A. F.</dc:creator>
<dc:creator>Moseki, R.</dc:creator>
<dc:creator>Abhimanyu, A.</dc:creator>
<dc:creator>Plessis, N. D.</dc:creator>
<dc:creator>Blackburn, J.</dc:creator>
<dc:creator>Mhlanga, M. M.</dc:creator>
<dc:creator>Kahn, C. R.</dc:creator>
<dc:creator>Brombacher, F.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Parihar, S. P.</dc:creator>
<dc:date>2025-05-23</dc:date>
<dc:identifier>doi:10.1101/2025.05.19.653976</dc:identifier>
<dc:title><![CDATA[Protein Kinase C δ: a critical hub regulating macrophage immunomodulatory functions during Mycobacterium tuberculosis infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.20.655037v1?rss=1">
<title>
<![CDATA[
Substrate heterogeneity promotes cancer cell dissemination 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.20.655037v1?rss=1"
</link>
<description><![CDATA[
While tumor malignancy has been extensively studied under the prism of genetic and epigenetic heterogeneity, tumor cell states also critically depend on reciprocal interactions with the microenvironment. This raises the hitherto untested possibility that heterogeneity of the untransformed tumor stroma can actively fuel malignant progression. As biological heterogeneity is inherently difficult to control, we adopted a reductionist approach and let tumor cells invade micro-engineered environments harboring obstacles with precision-controlled geometry. We find that not only the presence of obstacles, but more surprisingly their spatial disorder, causes a drastic shift from a collective to a single-cell mode of invasion - comparable in strength to cadherin loss. Combining live-imaging and perturbation experiments with minimal biophysical modeling, we demonstrate that cell detachments result both from local geometrical constraints and a global integration of spatial disorder over time. We show that different types of microenvironments map onto different universality classes of invasion dynamics - homogeneous substrates follow Kardar-Parisi-Zhang (KPZ) scaling, while disordered ones exhibit exponents consistent with KPZ with quenched disorder (KPZq). Our findings highlight generic physical principles for how the mode of cancer cell invasion depends on environmental heterogeneity, with potential implications to understand tumor evolution in vivo.
]]></description>
<dc:creator>Dunajova, Z.</dc:creator>
<dc:creator>Tasciyan, S.</dc:creator>
<dc:creator>Majek, J.</dc:creator>
<dc:creator>Merrin, J.</dc:creator>
<dc:creator>Sahai, E.</dc:creator>
<dc:creator>Sixt, M.</dc:creator>
<dc:creator>Hannezo, E.</dc:creator>
<dc:date>2025-05-23</dc:date>
<dc:identifier>doi:10.1101/2025.05.20.655037</dc:identifier>
<dc:title><![CDATA[Substrate heterogeneity promotes cancer cell dissemination]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.15.654267v1?rss=1">
<title>
<![CDATA[
CRITICAL ROLE OF ADULT-BORN DENTATE GRANULE NEURONS IN PATTERN COMPLETION 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.15.654267v1?rss=1"
</link>
<description><![CDATA[
The dentate gyrus contains neurons generated during development and adulthood, yet their distinct roles remain unclear. Using a transgenic mouse model, we show that adult-born neurons contribute to pattern completion, essential for episodic memory. This has implications for aging, PTSD, and addiction, where excessive pattern completion leads to maladaptive memories. Targeting ABNs could offer new therapeutic strategies to regulate memory processes and improve mental health.
]]></description>
<dc:creator>roullet, p.</dc:creator>
<dc:creator>Koehl, M.</dc:creator>
<dc:creator>Delage, P.</dc:creator>
<dc:creator>Guillemot, F.</dc:creator>
<dc:creator>Abrous, N.</dc:creator>
<dc:date>2025-05-20</dc:date>
<dc:identifier>doi:10.1101/2025.05.15.654267</dc:identifier>
<dc:title><![CDATA[CRITICAL ROLE OF ADULT-BORN DENTATE GRANULE NEURONS IN PATTERN COMPLETION]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.15.654179v1?rss=1">
<title>
<![CDATA[
The blood-brain barrier regulates brain tumour growth specifically via the SLC36 amino acid transporter Pathetic in Drosophila 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.15.654179v1?rss=1"
</link>
<description><![CDATA[
Tumours adapt their metabolism to sustain increased proliferation, rendering them particularly vulnerable to fluctuations in nutrient availability. However, the role of the tumour microenvironment in modulating sensitivity to nutrient restriction (NR) remains poorly understood. Using a Drosophila brain dedifferentiation neural stem cell (NSC) tumour model induced by Prospero (Pros) inhibition, we show that tumour sensitivity to NR is governed by the perineural glial (PG) cells of the blood-brain barrier (BBB), a major component of the glial niche surrounding the tumour. We identify the SLC36 amino acid transporter Pathetic (Path) as a crucial regulator of nutrient sensitivity. Under NR, while wildtype buffers against low nutrient levels by upregulating Path, tumour glia downregulate Path. Furthermore, Path is specifically required by the tumour (but not wildtype) PG; its downregulation causes reduced proliferation of PG cells and, in turn, restricts NSC tumour growth. Path influences PG proliferation via the mTor-S6K pathway, and its expression is controlled by Ilp6 levels and the Insulin/PI3K pathway. Overexpression of Path is sufficient to counteract the inhibitory effects of NR on tumour growth. These findings suggest that Path levels at the BBB play a key role in determining tumour sensitivity to NR.
]]></description>
<dc:creator>Dong, Q.</dc:creator>
<dc:creator>AlvarezOchoa, E.</dc:creator>
<dc:creator>Kosakamoto, H.</dc:creator>
<dc:creator>Obata, F.</dc:creator>
<dc:creator>Alexandre, C.</dc:creator>
<dc:creator>Cheng, L.</dc:creator>
<dc:date>2025-05-20</dc:date>
<dc:identifier>doi:10.1101/2025.05.15.654179</dc:identifier>
<dc:title><![CDATA[The blood-brain barrier regulates brain tumour growth specifically via the SLC36 amino acid transporter Pathetic in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.06.658228v1?rss=1">
<title>
<![CDATA[
A closed feedback between tissue phase transitions and morphogen gradients drives patterning dynamics 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.06.658228v1?rss=1"
</link>
<description><![CDATA[
During development mechanochemical cues in the cell microenvironment are translated into signalling to drive cell fate decisions. As cells differentiate collectively, it raises the question of how tissue-level properties affect instructive cues of decision-making. Here, we show that a tissue rigidity phase transition guides patterning by tuning the length-scales and time-scales of morphogen signalling. By combining rigidity percolation theory, reaction-diffusion modelling, quantitative imaging, optogenetics and single-cell transcriptomics in zebrafish, we uncover dynamical global tissue rigidity patterns that actively shape the Nodal morphogen gradient by restricting ligand dispersal and accelerating its signalling activity. In this self-generated mechanism, Nodal, besides driving meso-endoderm fate specification, increases cell-cell adhesion strength via regulating planar cell polarity genes. Once adhesion strength reaches a critical point, it triggers a rigidity transition which collapses tissue porosity. The abrupt tissue reorganisation negatively feeds back on Nodal signalling impacting both its length-scales, by limiting Nodal diffusivity, and its time-scales, by speeding up the expression of its antagonist Lefty, thereby ensuring timely signal termination and robust patterning. Overall, we reveal how emergent properties set the spatiotemporal dynamics of morphogen gradients, uncovering macroscopic mechanisms of pattern formation.
]]></description>
<dc:creator>Autorino, C.</dc:creator>
<dc:creator>Khoromskaia, D.</dc:creator>
<dc:creator>Harari, L.</dc:creator>
<dc:creator>Floris, E.</dc:creator>
<dc:creator>Booth, H.</dc:creator>
<dc:creator>Pallares-Cartes, C.</dc:creator>
<dc:creator>Petrasiunaite, V.</dc:creator>
<dc:creator>Dorrity, M.</dc:creator>
<dc:creator>Corominas-Murtra, B.</dc:creator>
<dc:creator>Hadjivasiliou, Z.</dc:creator>
<dc:creator>Petridou, N.</dc:creator>
<dc:date>2025-06-06</dc:date>
<dc:identifier>doi:10.1101/2025.06.06.658228</dc:identifier>
<dc:title><![CDATA[A closed feedback between tissue phase transitions and morphogen gradients drives patterning dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.06.657372v1?rss=1">
<title>
<![CDATA[
MRTF-dependent cytoskeletal dynamics drive efficient cell cycle progression 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.06.657372v1?rss=1"
</link>
<description><![CDATA[
Serum response factor (SRF) and its cofactors, Myocardin-related transcription factors A/B (MRTF-A/B), regulate transcription of numerous cytoskeletal structural and regulatory genes, and most MRTF/SRF inactivation phenotypes reflect deficits in cytoskeletal dynamics. We show that MRTF-SRF activity is required for effective proliferation of both primary and immortalised fibroblast and epithelial cells. Cells lacking the MRTFs or SRF proliferate very slowly, express elevated levels of SASP factors and SA-{beta}-galactosidase activity, and inhibit proliferation of co-cultured primary wildtype cells. They exhibit decreased levels of CDK1 and CKS2 proteins, and elevated levels of CDK inhibitors, usually CDKN1B/p27. These phenotypes, which can be fully reversed by re-expression of MRTF-A, are also seen in wildtype cells arrested by serum deprivation. Moreover, in wildtype cells direct interference with cytoskeletal dynamics through inhibition of ROCKs or Myosin ATPase induces a similar proliferative defect to that seen in MRTF-null cells. MRTF-null cells exhibit multiple cytoskeletal defects, and markedly reduced contractility. We propose that MRTF-SRF driven cytoskeletal dynamics and contractility are essential for operation of the pro-proliferative signal provided by cell-substrate adhesion.
]]></description>
<dc:creator>Nielsen, J. C.</dc:creator>
<dc:creator>Benito-Jardon, M.</dc:creator>
<dc:creator>Petrela, N. C.</dc:creator>
<dc:creator>Diring, J.</dc:creator>
<dc:creator>Bellamy, S.</dc:creator>
<dc:creator>Treisman, R.</dc:creator>
<dc:date>2025-06-07</dc:date>
<dc:identifier>doi:10.1101/2025.06.06.657372</dc:identifier>
<dc:title><![CDATA[MRTF-dependent cytoskeletal dynamics drive efficient cell cycle progression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.06.658331v1?rss=1">
<title>
<![CDATA[
Multimodal profiling unveils a reversible basal-like breast cancer cell state resistant to AKT inhibition 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.06.658331v1?rss=1"
</link>
<description><![CDATA[
The PI3K/AKT/mTOR pathway is central to cell metabolism and growth. However, pharmacological inhibition of the pathway is not uniformly effective across cancer types, or even within a single cancer model. In this study, we leverage oblique plane microscopy of triple negative breast cancer organoids, as well as lineage tracing to uncover a source of heterogeneity. Non-genetic resistance to AKT inhibition is associated with basal cell features of normal breast epithelium and the master transcription factor of basal cell state, {Delta}Np63, is sufficient to confer resistance. Cells can transition between states within four weeks and therefore, AKT inhibition only delays tumour growth, with tumours rich in KRT14+ cells resulting. Thus, under selection, triple negative breast cancer exploits a repertoire of cell states inherent to the breast.
]]></description>
<dc:creator>Ratcliffe, C. D. H.</dc:creator>
<dc:creator>Sparks, H.</dc:creator>
<dc:creator>Boezio, G. L. M.</dc:creator>
<dc:creator>Alexandrov, Y.</dc:creator>
<dc:creator>Jenkins, R. P.</dc:creator>
<dc:creator>Le Marois, A.</dc:creator>
<dc:creator>Soro-Barrio, P.</dc:creator>
<dc:creator>Lee, R.</dc:creator>
<dc:creator>Strohbuecker, S.</dc:creator>
<dc:creator>Joshi, S.</dc:creator>
<dc:creator>Ellis, J. K.</dc:creator>
<dc:creator>Fortier, A.-M.</dc:creator>
<dc:creator>Gustafsson, N.</dc:creator>
<dc:creator>Cunha, A. C.</dc:creator>
<dc:creator>Park, M.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Serio, A.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Riddell, A.</dc:creator>
<dc:creator>Barry, S. T.</dc:creator>
<dc:creator>Dunsby, C.</dc:creator>
<dc:creator>Sahai, E.</dc:creator>
<dc:date>2025-06-07</dc:date>
<dc:identifier>doi:10.1101/2025.06.06.658331</dc:identifier>
<dc:title><![CDATA[Multimodal profiling unveils a reversible basal-like breast cancer cell state resistant to AKT inhibition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.11.659158v1?rss=1">
<title>
<![CDATA[
Cross-species comparative connectomics reveals the evolution of an olfactory circuit 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.11.659158v1?rss=1"
</link>
<description><![CDATA[
Animal behavioural diversity ultimately stems from variation in neural circuitry, yet how central neural circuits evolve remains poorly understood. Studies of neural circuit evolution often focus on a few elements within a network. However, addressing fundamental questions in evolutionary neuroscience, such as whether some elements are more evolvable than others, requires a more global and unbiased approach. Here, we used synapse-level comparative connectomics to examine how an entire olfactory circuit evolves. We compared the full antennal lobe connectome of the larvae of two closely related Drosophila species, D. melanogaster and D. erecta, which differ in their ecological niches and odour-driven behaviours. We found that evolutionary change is unevenly distributed across the network. Some features, including neuron types, neuron numbers and interneuron-to-interneuron connectivity, are highly conserved. These conserved elements delineate a core circuit blueprint presumably required for fundamental olfactory processing. Superimposed on this scaffold, we find rewiring changes that mirror each species ecologies, including a systematic shift in the excitation-to-inhibition balance in the feedforward pathways. We further show that some neurons have changed more than others, and that even within individual neurons some synaptic elements remain conserved while others display major species-specific changes, suggesting evolutionary hot-spots within the circuit. Our findings reveal constrained and adaptable elements within olfactory networks, and establish a framework for identifying general principles in the evolution of neural circuits underlying behaviour.
]]></description>
<dc:creator>Roberts, R.</dc:creator>
<dc:creator>Giez, C.</dc:creator>
<dc:creator>Dhawan, S.</dc:creator>
<dc:creator>Pang, S.</dc:creator>
<dc:creator>Randel, N.</dc:creator>
<dc:creator>Zhiyuan, L.</dc:creator>
<dc:creator>Gong, H.</dc:creator>
<dc:creator>Dekens, L.</dc:creator>
<dc:creator>DiFrisco, J.</dc:creator>
<dc:creator>Xu, C. S.</dc:creator>
<dc:creator>Hess, H.</dc:creator>
<dc:creator>Zlatic, M.</dc:creator>
<dc:creator>Cardona, A.</dc:creator>
<dc:creator>Prieto-Godino, L. L.</dc:creator>
<dc:date>2025-06-11</dc:date>
<dc:identifier>doi:10.1101/2025.06.11.659158</dc:identifier>
<dc:title><![CDATA[Cross-species comparative connectomics reveals the evolution of an olfactory circuit]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.12.659257v1?rss=1">
<title>
<![CDATA[
SARS-CoV-2 envelope protein induces LC3 lipidation via the V-ATPase-ATG16L1 axis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.12.659257v1?rss=1"
</link>
<description><![CDATA[
Coronaviruses encode envelope (E), a structural component of the virion that is important for assembly and egress. E has proton channel activity that prevents premature rearrangement of the spike glycoprotein as virions encounter acidic compartments as they exit the cell. How infected cells respond to this pH disruption during coronavirus infection is unknown. Here we show that SARS-CoV-2 E ion channel activity triggers the proton pump V-ATPases to recruit the ATG16L1 complex during infection. This results in ATG8 molecules such as LC3B decorating perturbed compartments. This recruitment of autophagy machinery does not inhibit viral replication, rather SARS-CoV-2 exploits this response. Inhibition of the V-ATPase/ATG16L1 interaction using the Salmonella effector SopF inhibits SARS-CoV-2 replication. Careful distinction between use of the autophagic machinery from canonical macroautophagy is required in order to better understand coronavirus replication and for rational targeting of any potential host-directed therapies.
]]></description>
<dc:creator>Figueras-Novoa, C.</dc:creator>
<dc:creator>Timimi, L. J.</dc:creator>
<dc:creator>Marcassa, E.</dc:creator>
<dc:creator>Taveira-Marques, R.</dc:creator>
<dc:creator>Adams, L.</dc:creator>
<dc:creator>Jiang, M.</dc:creator>
<dc:creator>Wu, M. Y.</dc:creator>
<dc:creator>Montaner, B.</dc:creator>
<dc:creator>Ng, K.</dc:creator>
<dc:creator>De Lorenzo, G.</dc:creator>
<dc:creator>Furnon, W.</dc:creator>
<dc:creator>Cowton, V. M.</dc:creator>
<dc:creator>Upfold, N.</dc:creator>
<dc:creator>Kassiotis, G.</dc:creator>
<dc:creator>Harvey, R.</dc:creator>
<dc:creator>Patel, A. H.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>Ulferts, R.</dc:creator>
<dc:creator>Beale, R.</dc:creator>
<dc:date>2025-06-13</dc:date>
<dc:identifier>doi:10.1101/2025.06.12.659257</dc:identifier>
<dc:title><![CDATA[SARS-CoV-2 envelope protein induces LC3 lipidation via the V-ATPase-ATG16L1 axis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.13.659524v1?rss=1">
<title>
<![CDATA[
Overnight circuit remodelling drives juvenile alloparental care 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.13.659524v1?rss=1"
</link>
<description><![CDATA[
Parental care is critical for the survival of altricial young and is mediated by neural circuits that are well characterised in adult rodents. Although adults can exhibit caregiving even before becoming parents, the developmental origins of this so-called alloparental behaviour remain unclear. Here, we show that alloparental behaviour in mice emerges abruptly between postnatal day (P)14 and 15, independently of prior social experience. This behavioural transition coincides with the onset of pup-specific activity in galanin-expressing medial preoptic area (MPOAGal) neurons, which are essential for parental behaviour in adulthood. Chemogenetic silencing of MPOAGal neurons abolishes caregiving in juveniles, suggesting that similar circuits control parenting across life stages. Viral trans-synaptic tracing and whole-cell recordings reveal extensive input remodelling of MPOAGal neurons between P14 and 15, marking a rapid transition from a highly connected, immature network to a sparser, adult-like circuit configuration. We identify microglia as key mediators of this process, as their ablation prevents both synaptic reorganisation and the emergence of alloparenting. Together, these findings uncover a previously unrecognised, microglia-dependent developmental switch that enables caregiving in juveniles through rapid circuit reconfiguration.
]]></description>
<dc:creator>Jamieson, B. B.</dc:creator>
<dc:creator>Chen, M. X.</dc:creator>
<dc:creator>Liang, S.</dc:creator>
<dc:creator>El Rasheed, L. S. H.</dc:creator>
<dc:creator>Chattey, G. M. K.</dc:creator>
<dc:creator>Kohl, J.</dc:creator>
<dc:date>2025-06-15</dc:date>
<dc:identifier>doi:10.1101/2025.06.13.659524</dc:identifier>
<dc:title><![CDATA[Overnight circuit remodelling drives juvenile alloparental care]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.12.659321v1?rss=1">
<title>
<![CDATA[
VISIBLE: An imaging-driven system for sampling, biofabrication, and manipulation of complex biological models 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.12.659321v1?rss=1"
</link>
<description><![CDATA[
Complex in vitro models (CIVMs), including organoids, spheroids, and bioprinted constructs, have emerged as powerful platforms for recapitulating human tissue architecture and function. However, their inherent heterogeneity and dynamic nature pose significant challenges for standardization, reproducibility, and real-time manipulation. Here, we present VISIBLE (Versatile Imaging-Based Interactive Sampling and Bioprinting System), a modular, imaging-guided platform that integrates real-time monitoring with automated manipulation and 3D bioprinting that addresses these challenges. VISIBLE employs a dual-axis system comprising independently controlled tool-heads and microscope stages, enabling precise spatiotemporal interventions within live cultures. We demonstrate the system capabilities across a range of applications, including morphology- and function-based sampling of organoids and neurospheres, interactive 3D bioprinting, serial scratch assays, and real-time cell-cell interaction studies. Furthermore, we illustrate the system utility in translational contexts through the selective sampling and implantation of patient-derived xenograft organoids. VISIBLE supports long-term culture within an integrated incubation environment and accommodates interchangeable tool-heads for scalable, high-throughput workflows. By enabling closed-loop, feedback-controlled experimentation, VISIBLE addresses some of the critical limitations in existing CIVM platforms and offers a new versatile solution for a wide range of biomedical research applications. This work represents a significant step toward the development of autonomous experimental systems for complex tissue modelling and preclinical investigation.
]]></description>
<dc:creator>Joshi, S.</dc:creator>
<dc:creator>Moreno-Gonzalez, C.</dc:creator>
<dc:creator>Suklai, P.</dc:creator>
<dc:creator>Carraro, E.</dc:creator>
<dc:creator>Ratcliffe, C. D. H.</dc:creator>
<dc:creator>Boezio, G. L. M.</dc:creator>
<dc:creator>Konstantinou, G.</dc:creator>
<dc:creator>Hagemann, C.</dc:creator>
<dc:creator>Kavanagh, T.</dc:creator>
<dc:creator>Ameer-Beg, S.</dc:creator>
<dc:creator>Imbert, A.</dc:creator>
<dc:creator>Sahai, E.</dc:creator>
<dc:creator>Tedesco, F. S.</dc:creator>
<dc:creator>Serio, A.</dc:creator>
<dc:date>2025-06-17</dc:date>
<dc:identifier>doi:10.1101/2025.06.12.659321</dc:identifier>
<dc:title><![CDATA[VISIBLE: An imaging-driven system for sampling, biofabrication, and manipulation of complex biological models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.13.659133v1?rss=1">
<title>
<![CDATA[
Anti-activin treatment increases T cell infiltration in breast and pancreatic tumours and promotes survival in a SMAD4-null mouse pancreatic cancer model 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.13.659133v1?rss=1"
</link>
<description><![CDATA[
Activin A and B share the same downstream signalling pathway (activation of SMAD2/3) as TGF-{beta} and consequently elicit many of the same functional responses as TGF-{beta}, including immune suppression, activation of cancer-associated fibroblasts (CAFs) and extracellular matrix production and remodelling. However, activins role in tumourigenesis has been relatively overlooked compared to TGF-{beta}s. We generated and characterized a dual specificity human antibody that recognizes both activin A and B and compared its activity in syngeneic mouse models of breast cancer and pancreatic ductal adenocarcinoma (PDAC) with an activin A-specific antibody. We demonstrate that activin A and B are central to the function of CAFs and therapeutic inhibition of activin results in a reduction of collagen rich desmoplastic barriers, enabling the infiltration of cytotoxic T cells. This is correlated with an upregulation of the T cell chemoattractant CXCL10, which is normally repressed by activin signalling. Interestingly, despite greater T cell infiltration, activin A inhibition resulted in poorer survival in the KPC mouse model of PDAC and slightly larger tumours in the breast cancer model, indicating a tumour suppressive role of activin A-rich CAFs. Strikingly, however, treatment with the same anti-activin A antibody of PDAC tumours where SMAD4 is deleted in the tumour cells, resulted in increased survival, which was potentiated with additional treatment with immune checkpoint inhibitors. These results suggest that anti-activin therapy has potential for the cohort of PDAC patients exhibiting inactivation of SMAD4.
]]></description>
<dc:creator>McCluney, S.</dc:creator>
<dc:creator>Park, D.</dc:creator>
<dc:creator>Miller, D. S.</dc:creator>
<dc:creator>Mehic, M.</dc:creator>
<dc:creator>Bloxham, R. D.</dc:creator>
<dc:creator>De Vries, E.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Boeing, S.</dc:creator>
<dc:creator>D'Antonio, L.</dc:creator>
<dc:creator>Wieteska, L.</dc:creator>
<dc:creator>Pyeatt, G.</dc:creator>
<dc:creator>Strohbuecker, S.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Maia De Oliveira, T.</dc:creator>
<dc:creator>Wilcockson, R.</dc:creator>
<dc:creator>Hyvonen, M.</dc:creator>
<dc:creator>Hunt, J.</dc:creator>
<dc:creator>Hill, C. S.</dc:creator>
<dc:date>2025-06-17</dc:date>
<dc:identifier>doi:10.1101/2025.06.13.659133</dc:identifier>
<dc:title><![CDATA[Anti-activin treatment increases T cell infiltration in breast and pancreatic tumours and promotes survival in a SMAD4-null mouse pancreatic cancer model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.17.659946v1?rss=1">
<title>
<![CDATA[
The evolution of gene regulatory programs controlling gonadal development in primates 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.17.659946v1?rss=1"
</link>
<description><![CDATA[
Sex-determining pathways produce dimorphic gonads (ovaries and testes), yet the gene regulatory programs governing gonadogenesis and their evolution in primates remain little explored. Here we report evolutionary analyses of transcriptome and chromatin accessibility data of male and female human, marmoset (New World monkey), and mouse gonadal cells spanning key prenatal stages. We find that the two primates and mouse share similar X chromosome expression dynamics, including X chromosome reactivation (XCR), and that in Klinefelter syndrome (XXY) testes, germ cells undergo female-like XCR and escape of X inactivation. New male-specific regulatory regions have emerged progressively during mammalian evolution, especially on the X following sex chromosome origination. Further analyses revealed that male-specific regulatory regions evolved faster than female-specific ones in both supporting and pre-meiotic germ cells. However, female meiotic germ cells show even higher rates of molecular evolution and exhibit a permissive chromatin state that facilitates the birth of new genes, thus resembling their adult spermatogenic counterparts. Finally, we traced both conserved and species-specific gene expression trajectories across the three mammals, uncovering candidate genes for disorders of sex development that are typically central to cell-type-specific regulatory networks. Together, our study unveils both ancestral mammalian and recently evolved gene regulatory programs that control human- and primate-specific aspects of gonadal development in both sexes.
]]></description>
<dc:creator>Trost, N.</dc:creator>
<dc:creator>Fallahshahroudi, A.</dc:creator>
<dc:creator>Sarropoulos, I.</dc:creator>
<dc:creator>Schneider, C.</dc:creator>
<dc:creator>Schmidt, J.</dc:creator>
<dc:creator>Mbengue, N.</dc:creator>
<dc:creator>Wolff, E.</dc:creator>
<dc:creator>Drummer, C.</dc:creator>
<dc:creator>Frömel, R.</dc:creator>
<dc:creator>Lisgo, S.</dc:creator>
<dc:creator>Murat, F.</dc:creator>
<dc:creator>Sepp, M.</dc:creator>
<dc:creator>Cardoso-Moreira, M.</dc:creator>
<dc:creator>Behr, R.</dc:creator>
<dc:creator>Kaessmann, H.</dc:creator>
<dc:date>2025-06-19</dc:date>
<dc:identifier>doi:10.1101/2025.06.17.659946</dc:identifier>
<dc:title><![CDATA[The evolution of gene regulatory programs controlling gonadal development in primates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.22.660928v1?rss=1">
<title>
<![CDATA[
Advanced human iPSC-based modelling of LMNA-related congenital muscular dystrophy enables development of targeted genetic therapies for muscle laminopathies 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.22.660928v1?rss=1"
</link>
<description><![CDATA[
LMNA-related congenital muscular dystrophy (L-CMD) is amongst the most severe forms of laminopathies, which are diseases caused by pathogenic variants in the LMNA gene. LMNA encodes the proteins Lamin A and C, which assemble with Lamin B1 and B2 to form the nuclear lamina: a meshwork providing structural stability to the nucleus that also regulates chromatin organisation and gene expression. Research into L-CMD mechanisms and therapies is hindered by lack of humanised, tissue-specific models that accurately recapitulate disease phenotypes. We previously reported that LMNA-mutant induced pluripotent stem cell (iPSC)-derived skeletal muscle cells have nuclear shape abnormalities and Lamin A/C protein mislocalisation. Here, we expand the selection of L-CMD patient- derived iPSCs and validate disease-associated readouts using a transgene-free based protocol which more accurately mimics skeletal myogenesis. Results showed no overt defects in developmental myogenesis, but recapitulation of pathological nuclear shape abnormalities in 2D and 3D cultures, nuclear envelope protein mislocalisation and transcriptomic alterations across multiple pathogenic LMNA variants. We then utilised this platform to assess LMNA gene editing strategies. CRISPR-based exon removal generated stable RNA and protein Lamin A/C species, without significant normalisation of nuclear morphological phenotypes or transcriptomic profile. Conversely, precise editing of the same mutation showed complete reversal of disease-associated nuclear morphometrics, alongside normalisation of the pro-inflammatory transcriptomic signature. Our data provide the foundation for a humanised in vitro disease and therapy modelling platform for this complex and severe muscle disorder.

GRAPHICAL ABSTRACT

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=81 SRC="FIGDIR/small/660928v2_ufig1.gif" ALT="Figure 1">
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org.highwire.dtl.DTLVardef@134276forg.highwire.dtl.DTLVardef@108f44forg.highwire.dtl.DTLVardef@dfa04eorg.highwire.dtl.DTLVardef@1956a90_HPS_FORMAT_FIGEXP  M_FIG C_FIG HIGHLIGHTSO_LILMNA-mutant iPSCs undergo efficient skeletal myogenesis upon transgene-free, small molecule-based lineage-directed differentiation
C_LIO_LIL-CMD iPSCs recapitulate hallmark disease-associated nuclear phenotypes and show a pro-inflammatory transcriptional profile
C_LIO_LIDisease modelling platforms based on iPSC-derived skeletal muscle cells enable comparative testing of gene editing strategies
C_LIO_LICRISPR-edited L-CMD iPSC-derived myogenic cells show amelioration of disease-associated readouts
C_LI
]]></description>
<dc:creator>Moore, D. P.</dc:creator>
<dc:creator>Steele-Stallard, H. B.</dc:creator>
<dc:creator>Pinton, L.</dc:creator>
<dc:creator>Lionello, V. M.</dc:creator>
<dc:creator>Rossi, L.</dc:creator>
<dc:creator>Aghaeipour, A.</dc:creator>
<dc:creator>Jalal, S.</dc:creator>
<dc:creator>Wong, C. T. Y.</dc:creator>
<dc:creator>Clara-Hwang, A.</dc:creator>
<dc:creator>Bonne, G.</dc:creator>
<dc:creator>Zammit, P. S.</dc:creator>
<dc:creator>Tedesco, F. S.</dc:creator>
<dc:date>2025-06-23</dc:date>
<dc:identifier>doi:10.1101/2025.06.22.660928</dc:identifier>
<dc:title><![CDATA[Advanced human iPSC-based modelling of LMNA-related congenital muscular dystrophy enables development of targeted genetic therapies for muscle laminopathies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.23.661120v1?rss=1">
<title>
<![CDATA[
Sensitizing Immune-Refractory Ovarian Tumors via p53 Mutation-Tailored Immunotherapy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.23.661120v1?rss=1"
</link>
<description><![CDATA[
High-grade serous ovarian cancer demonstrates limited responsiveness to immune checkpoint inhibitors, owing in part to immunosuppressive environments shaped by nearly universal p53 aberrations. Utilizing an immunocompetent mouse model and individual p53 mutations, we identified a dependence of the p53-R270H mutation (equivalent of human R273H) on regulatory T cells (Tregs) and the PD-1/PD-L1 axis. Analysis of patient datasets associated R273H with elevated levels of two p53 targets, PD-L1 and amphiregulin (AREG), a Tregs growth factor. In contrast to p53-R172H tumors, where there was limited activity, dual antibody therapy targeting AREG and PD-L1 selectively and effectively inhibited R270H tumors. This involved polarization toward M1 macrophages, infiltration of CD8+ T cells, diminished Ly6G+ neutrophils and downregulation of interleukin-4. In patient-derived R273C organoids, the combination treatment reduced the CD4/CD8 ratio. This study is the first to establish a mutation-tailored therapeutic approach that leverages the capacity of p53 to modulate immunosuppressive mechanisms.
]]></description>
<dc:creator>Chatterjee, R.</dc:creator>
<dc:creator>Simoni-Nieves, A.</dc:creator>
<dc:creator>Truong, A.</dc:creator>
<dc:creator>Lindzen, M.</dc:creator>
<dc:creator>Ozmen, F.</dc:creator>
<dc:creator>Cherry, C.</dc:creator>
<dc:creator>Zwicky, P.</dc:creator>
<dc:creator>Mukherjee, S.</dc:creator>
<dc:creator>Selvadurai, B.-R.</dc:creator>
<dc:creator>Salame, T. M.</dc:creator>
<dc:creator>Gupta, N.</dc:creator>
<dc:creator>Giri, S.</dc:creator>
<dc:creator>Kramarski, L.</dc:creator>
<dc:creator>Avraham, Y.</dc:creator>
<dc:creator>Weizman, E.</dc:creator>
<dc:creator>Ozmen, T.</dc:creator>
<dc:creator>Noronha, A.</dc:creator>
<dc:creator>Chakrabarti, P.</dc:creator>
<dc:creator>Ramesh-Kumar, D.</dc:creator>
<dc:creator>Downward, J.</dc:creator>
<dc:creator>Dahan, R.</dc:creator>
<dc:creator>Amit, I.</dc:creator>
<dc:creator>Velculescu, V.</dc:creator>
<dc:creator>Brenton, J.</dc:creator>
<dc:creator>Mills, G.</dc:creator>
<dc:creator>Oren, M.</dc:creator>
<dc:creator>Yarden, Y.</dc:creator>
<dc:date>2025-06-27</dc:date>
<dc:identifier>doi:10.1101/2025.06.23.661120</dc:identifier>
<dc:title><![CDATA[Sensitizing Immune-Refractory Ovarian Tumors via p53 Mutation-Tailored Immunotherapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.24.661269v1?rss=1">
<title>
<![CDATA[
The adaptive molecular landscape of reprogrammed telomeric sequences 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.24.661269v1?rss=1"
</link>
<description><![CDATA[
Telomeric sequences vary across the tree of life and intimately co-evolve with telomere-binding protein complexes. However, the molecular mechanisms allowing organisms to adapt to new telomeric sequences are difficult to gauge from extant species. Here, we reprogrammed multiple yeast lines to human-like telomeric repeats to unveil their molecular and fitness response to novel telomeres. Initially, the exchange of telomere sequences resulted in genome instability, proteome remodelling and severe fitness decline. However, adaptive evolution experiments selected for repeated mutations that drove adaptation to the humanized telomeres. These consisted of the recurrent amplification of the telomere-binding protein TBF1, by complex aneuploidies, or in repeated mutations that attenuate the DNA damage response. Overall, our results outline a response that defines the adaptive molecular landscape to novel telomeric sequences.
]]></description>
<dc:creator>D'Angiolo, M.</dc:creator>
<dc:creator>Barre, B. P.</dc:creator>
<dc:creator>Khaiwal, S.</dc:creator>
<dc:creator>Muenzner, J.</dc:creator>
<dc:creator>Hallin, J.</dc:creator>
<dc:creator>De Chiara, M.</dc:creator>
<dc:creator>Tellini, N.</dc:creator>
<dc:creator>Warringer, J.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:creator>Gilson, E.</dc:creator>
<dc:creator>Liti, G.</dc:creator>
<dc:date>2025-06-27</dc:date>
<dc:identifier>doi:10.1101/2025.06.24.661269</dc:identifier>
<dc:title><![CDATA[The adaptive molecular landscape of reprogrammed telomeric sequences]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.07.01.662575v1?rss=1">
<title>
<![CDATA[
An optimised computational approach for the identification of somatic structural variants in cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.07.01.662575v1?rss=1"
</link>
<description><![CDATA[
Structural variants play a critical role in tumorigenesis. At present, these events are most commonly identified using short-read whole-genome sequencing data, and a number of computational tools are available for this purpose. Consensus approaches have been used to improve precision, but may reduce sensitivity. The optimal number and combination of callers remains unclear, in part due to the lack of gold standard real-world datasets for validation. Here, we benchmark the performance of Delly, GRIDSS, LUMPY, Manta and SvABA, using a validation set of consensus calls from the Pan-Cancer Analysis of Whole Genomes Consortium. Manta showed the best standalone performance, identifying 88% of the validation set calls, and was included in all of the best-performing caller combinations. A consensus approach comprising Delly, GRIDSS, Manta and SvABA was selected as the optimum approach from those tested. We provide a NextFlow implementation of our optimised consensus approach as a resource for the cancer genomics community.
]]></description>
<dc:creator>Waise, S.</dc:creator>
<dc:creator>Mensah, N.</dc:creator>
<dc:creator>Lesluyes, T.</dc:creator>
<dc:creator>Demeulemeester, J.</dc:creator>
<dc:creator>Flanagan, A. M.</dc:creator>
<dc:creator>Pillay, N.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:date>2025-07-02</dc:date>
<dc:identifier>doi:10.1101/2025.07.01.662575</dc:identifier>
<dc:title><![CDATA[An optimised computational approach for the identification of somatic structural variants in cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.27.661680v1?rss=1">
<title>
<![CDATA[
Lonafarnib Partially Reverses Cardiac Senescence in Human and Mouse Progeria Models via Autophagy Activation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.27.661680v1?rss=1"
</link>
<description><![CDATA[
Hutchinson-Gilford Progeria Syndrome (HGPS), characterised by accelerated ageing, causes cardiovascular defects resembling aspects of cardiovascular ageing. We used human left ventricle cardiomyocytes (CMs) derived from HGPS-induced pluripotent stem cells (iPSCs), and their isogenic-corrected controls, to investigate HGPS-CM dysfunction and identify potential therapies. Our results revealed that HGPS-iPSC-CMs exhibit greater maturity and associated elevated oxidative stress compared to controls, which they could not contend with, leading to cellular senescence. Increased senescence was also observed in cardiac tissue from mouse and human physiologically-aged and HGPS individuals. Functionally, HGPS-iPSC-CMs showed dysregulated mitochondrial respiration and calcium handling. Amongst the six drugs tested, rapamycin and lonafarnib were the most effective against HGPS-cardiac phenotypes. Although lonafarnib raised safety concerns, it partially reverted the cardiac senescent phenotype by inducing cellular autophagy and decreasing progerin expression in progeroid mice. Our study supports the use of HGPS-iPSC-CMs to identify novel biomarkers and therapies for HGPS, and potentially cardiac physiological-ageing.
]]></description>
<dc:creator>Monteiro, L. M.</dc:creator>
<dc:creator>Pitrez, P. R.</dc:creator>
<dc:creator>Correira-Santos, S.</dc:creator>
<dc:creator>Dark, N.</dc:creator>
<dc:creator>Santinha, D.</dc:creator>
<dc:creator>Aveleira, C.</dc:creator>
<dc:creator>Townsend, M.</dc:creator>
<dc:creator>Taylor, J.</dc:creator>
<dc:creator>Nissan, X.</dc:creator>
<dc:creator>Sardao, V.</dc:creator>
<dc:creator>Cortes, L.</dc:creator>
<dc:creator>Mitter, R.</dc:creator>
<dc:creator>Marino, A.</dc:creator>
<dc:creator>Sousa-Soares, C.</dc:creator>
<dc:creator>Ribeiro, R.</dc:creator>
<dc:creator>Smith, J. C.</dc:creator>
<dc:creator>Logarinho, E.</dc:creator>
<dc:creator>Ori, A.</dc:creator>
<dc:creator>Bernardo, A. S.</dc:creator>
<dc:creator>Ferreira, L.</dc:creator>
<dc:date>2025-07-02</dc:date>
<dc:identifier>doi:10.1101/2025.06.27.661680</dc:identifier>
<dc:title><![CDATA[Lonafarnib Partially Reverses Cardiac Senescence in Human and Mouse Progeria Models via Autophagy Activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.30.662346v1?rss=1">
<title>
<![CDATA[
Metabolic control of enteroendocrine cell fate through a redox state sensor CtBP 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.30.662346v1?rss=1"
</link>
<description><![CDATA[
Enteroendocrine (EE) cells monitor the intestinal nutrient composition and consequently control organismal physiology through hormonal signaling. In addition to the immediate effects on hormone secretion, nutrients influence EE cell abundance by affecting the determination and maintenance of cell fate. EE cells are known to import and respond to dietary sugars, but how the sugar-induced changes in the intracellular metabolic state are sensed to control the immediate and long-term responses of EE cells, remains poorly understood. We report that the NADH binding transcriptional cofactor C-terminal binding protein (CtBP) acts at the interface between nutrient sensing and fate regulation of Drosophila larval EE cells, thus controlling organismal energy metabolism and survival on a high sugar diet. CtBP dimerization in EE cells is regulated through the redox balance of nicotinamide cofactors controlled by glycolysis and pentose phosphate pathway, allowing EE cells sense their internal metabolic state in response to sugar catabolism. CtBP interacts with the EE cell fate determining transcription factor Prospero through a conserved binding motif and binds to genomic targets controlling EE cell fate and size, such as components of Notch and insulin/mTOR pathways. Collectively, our findings uncover a modality where changes in intracellular redox state serve as an instructive signal to control EE cell function to globally control organismal homeostasis.

GRAPHICAL ABSTRACT

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=109 SRC="FIGDIR/small/662346v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Rovenko, B. M.</dc:creator>
<dc:creator>Moisio, K.</dc:creator>
<dc:creator>Biehler, C.</dc:creator>
<dc:creator>Girych, M.</dc:creator>
<dc:creator>Hallasaari, A.</dc:creator>
<dc:creator>Deniz, O.</dc:creator>
<dc:creator>Bluhm, S.</dc:creator>
<dc:creator>Viitanen, A.</dc:creator>
<dc:creator>Kokki, K.</dc:creator>
<dc:creator>Fabris, G.</dc:creator>
<dc:creator>Yang, Y.</dc:creator>
<dc:creator>Cracan, V.</dc:creator>
<dc:creator>Miguel-Aliaga, I.</dc:creator>
<dc:creator>Hietakangas, V.</dc:creator>
<dc:date>2025-07-02</dc:date>
<dc:identifier>doi:10.1101/2025.06.30.662346</dc:identifier>
<dc:title><![CDATA[Metabolic control of enteroendocrine cell fate through a redox state sensor CtBP]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.07.04.663185v1?rss=1">
<title>
<![CDATA[
Control of CDK activity and the cell cycle by CKS proteins 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.07.04.663185v1?rss=1"
</link>
<description><![CDATA[
The eukaryotic cell cycle is orchestrated by the activity of Cyclin-Dependent Kinases (CDKs)1. Although most mechanisms controlling CDK activity are well understood, one class of proteins - the CDK regulatory subunit (CKS) proteins - remain highly enigmatic. There is no generally accepted consistent molecular or functional characterisation of CKS in vivo, despite being essential across eukaryotes and implicated in a range of cell cycle processes and cancer2,3. Here, we provide a unifying framework for CKS function. We show in a single, genetically tractable system, the fission yeast, that CKS regulates the onset and progression of both S-phase and mitosis. We find that CKS modulates the phosphorylation of more than 200 CDK phosphosites in vivo, located on 133 substrate proteins. These are involved in diverse processes across the entire cell cycle, including DNA replication, chromosome condensation, the spindle assembly checkpoint, and the metaphase-anaphase transition. We demonstrate that CKS enhances CDK activity to drive phosphorylation of sites with low affinity for CDK. This acts both on specific substrates, likely by stabilizing CDK-substrate interactions, and through control of overall CDK activity, by regulating its interaction with Wee1 and Cdc25. Our findings establish CKS proteins as major multifaceted regulators of the cell cycle, operating through the global and local control of CDK activity.
]]></description>
<dc:creator>Curran, J. F.</dc:creator>
<dc:creator>Basu, S.</dc:creator>
<dc:creator>Auchynnikava, T.</dc:creator>
<dc:creator>Nurse, P.</dc:creator>
<dc:date>2025-07-07</dc:date>
<dc:identifier>doi:10.1101/2025.07.04.663185</dc:identifier>
<dc:title><![CDATA[Control of CDK activity and the cell cycle by CKS proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.07.10.664141v1?rss=1">
<title>
<![CDATA[
Kandinsky: enabling neighbourhood analysis of spatial omics data for functional insights on cell ecosystems 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.07.10.664141v1?rss=1"
</link>
<description><![CDATA[
Spatially resolved omics technologies enable investigation of how cells interact within their local environments or neighbourhoods directly in situ. Although a few computational methods have been developed to aid this analysis, significant limitations still exist in the way neighbourhoods are defined and exploited for downstream analyses. Here, we present Kandinsky, a computational tool that implements multiple approaches for neighbourhood identification, enabling high flexibility and versatility to address a variety of biological questions. Once identified, Kandinsky applies neighbourhoods for downstream studies, including proximity-based cell grouping for functional comparisons, spatial co-localisation and dispersion, and identification of hot and cold expression areas within the tissue. We apply Kandinsky to transcriptomic and proteomic data from different spatial technologies to showcase how it can reveal functional interactions between cells across multiple biological contexts.

Availability and implementationKandinsky is freely available as an R package at https://github.com/ciccalab/Kandinsky.
]]></description>
<dc:creator>Andrei, P.</dc:creator>
<dc:creator>Grieco, M.</dc:creator>
<dc:creator>Acha-Sagredo, A.</dc:creator>
<dc:creator>Dhami, P.</dc:creator>
<dc:creator>Fung, K.</dc:creator>
<dc:creator>Rodriguez-Justo, M.</dc:creator>
<dc:creator>Cereda, M.</dc:creator>
<dc:creator>Ciccarelli, F. D.</dc:creator>
<dc:date>2025-07-16</dc:date>
<dc:identifier>doi:10.1101/2025.07.10.664141</dc:identifier>
<dc:title><![CDATA[Kandinsky: enabling neighbourhood analysis of spatial omics data for functional insights on cell ecosystems]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.07.10.664086v1?rss=1">
<title>
<![CDATA[
A new material constant of DNA is the major determinant of looping dynamics 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.07.10.664086v1?rss=1"
</link>
<description><![CDATA[
DNA bending and looping is crucial for gene expression, packaging, and chromatin organisation, as well as the design of artificial nanomaterials and devices. But what determines how quickly DNA bends? While DNAs static flexibility is well-characterised by its persistence length, we lack an understanding of how quickly DNA responds to mechanical forces: remarkably current semiflexible polymer theory based on solvent dissipation underestimates spontaneous looping times by ~1000-fold. By analysing fluctuations of DNA several kilobases long and developing new theory for bending dissipation in semiflexible polymers, we show DNA bending dynamics cannot be explained by solvent friction alone and requires significant contributions from intramolecular friction. The theory defines a new material constant of DNA -- the bending friction, which we determine to be{zeta} B = 241 {+/-} 17 g nm3/ms. Strikingly, our measurement does not depend on the buffer ionic conditions. We predict bending friction will dominate DNA dynamics between {approx} 50 nm and 420 nm and significantly longer under external force. We show that mean first passage time calculations are greatly simplified when bending friction dominates and so using this constant, with no fitting parameters, we accurately predict the slow experimental spontaneous looping times. Our discovery of significant bending dissipation is unexpected as DNA has no obvious large (> kBT) internal energy barriers. The salt-independence of this dissipation also rules out long range electrostatic interactions as its origins. Instead our findings point to a complex local energy landscape for bending and a potential previously unappreciated role of water binding DNA constraining its local mobility. Our findings radically change our understanding of DNA dynamics and reveal DNA as a viscoelastic semiflexible polymer with dramatically slower dynamics compared to an ideal elastic rod. This work establishes bending friction as a fundamental material property that must underpin any model of DNA dynamics in biology, physics, and nanotechnology.
]]></description>
<dc:creator>Pobegalov, G.</dc:creator>
<dc:creator>Molodtsov, M.</dc:creator>
<dc:creator>Khatri, B. S.</dc:creator>
<dc:date>2025-07-16</dc:date>
<dc:identifier>doi:10.1101/2025.07.10.664086</dc:identifier>
<dc:title><![CDATA[A new material constant of DNA is the major determinant of looping dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.07.14.664731v1?rss=1">
<title>
<![CDATA[
Differential chromatin response to retinoic acid in neuroblastoma according to type of ATRX mutation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.07.14.664731v1?rss=1"
</link>
<description><![CDATA[
Neuroblastoma is a childhood cancer, arising in the developing sympathetic nervous system. Differentiation therapy with 13-cis-retinoic acid (RA) is routinely given to children with high-risk neuroblastoma in the minimal residual disease setting to prevent relapse, however there is little understanding of which patients benefit from RA therapy.

ATRX alterations are identified in 10% of high-risk neuroblastomas and associated with poor outcomes. The commonest type of ATRX alterations in neuroblastoma are in-frame multi-exon deletions, followed by nonsense mutations predicted to result in loss-of-function (ATRX LoF).

We treated paired ATRX wild-type and ATRX LoF neuroblastoma cell lines with RA and show that cells with ATRX LoF fail to upregulate direct RA target genes. Cells with ATRX LoF also show reduced chromatin accessibility at genes involved in differentiation and development following RA treatment. Conversely, neuroblastoma models with in-frame deletions mount a response to RA and show in-vitro sensitivity to RA. Taken together this shows that the mechanism of differentiation in ATRX-altered neuroblastoma depends on the type of ATRX alteration, with implications relating to both oncogenesis and therapeutic response.
]]></description>
<dc:creator>Lorenzi, F.</dc:creator>
<dc:creator>Shipley, M.</dc:creator>
<dc:creator>Deane, L.</dc:creator>
<dc:creator>Goldstone, R.</dc:creator>
<dc:creator>Tandon, V.</dc:creator>
<dc:creator>Martins da Costa, B.</dc:creator>
<dc:creator>Greenslade, K.</dc:creator>
<dc:creator>Barker, K.</dc:creator>
<dc:creator>Nemati, F.</dc:creator>
<dc:creator>Bellini, A.</dc:creator>
<dc:creator>Schleiermacher, G.</dc:creator>
<dc:creator>Chesler, L.</dc:creator>
<dc:creator>Guillemot, F.</dc:creator>
<dc:creator>George, S. L.</dc:creator>
<dc:date>2025-07-18</dc:date>
<dc:identifier>doi:10.1101/2025.07.14.664731</dc:identifier>
<dc:title><![CDATA[Differential chromatin response to retinoic acid in neuroblastoma according to type of ATRX mutation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.07.11.664267v1?rss=1">
<title>
<![CDATA[
A Paracrine-to-Autocrine Shunt of GREM1 Fuels Colorectal Cancer Metastasis via ACVR1C 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.07.11.664267v1?rss=1"
</link>
<description><![CDATA[
Tumor cells typically rely on paracrine stromal signals to guide malignant behaviors, but whether they acquire signaling autonomy to support metastasis remains unclear. We elucidate this in colorectal cancer (CRC) by uncovering a paracrine-to-autocrine shunt of Gremlin1 (GREM1), a canonical stromal-secreted antagonist of bone morphogenetic proteins (BMPs). We demonstrate that while GREM1 remains restricted to stromal cells in earlier-stage (I-III) CRC, its ectopic expression in tumor epithelium increases markedly in stage IV. Mechanistically, we identify ACVR1C as a novel, high-affinity epithelial receptor for GREM1. Their interaction activates SMAD2/3 signaling, which upregulates SNAI1 and GREM1, establishing a feedback loop that amplifies epithelial-mesenchymal transition (EMT). Disrupting this loop impairs CRC metastasis in vivo. Clinically, epithelial GREM1 or ACVR1C expression predicts metastasis and poor survival. These findings define a paradigm in which tumor cells hijack stromal GREM1 to establish a GREM1-ACVR1C autocrine loop that sustains EMT and metastasis, marking a shift toward signaling autonomy and revealing a targetable vulnerability in advanced CRC.
]]></description>
<dc:creator>Zhou, H.</dc:creator>
<dc:creator>Jin, Q.</dc:creator>
<dc:creator>Fu, Z.</dc:creator>
<dc:creator>Yang, Y.</dc:creator>
<dc:creator>Gao, Y.</dc:creator>
<dc:creator>Wang, N.</dc:creator>
<dc:creator>Zhao, B.</dc:creator>
<dc:creator>Gui, L.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Zhu, Z.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>He, Y.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Luo, S.</dc:creator>
<dc:creator>Fu, L.</dc:creator>
<dc:creator>Wu, X.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Shen, X.</dc:creator>
<dc:creator>Wang, T.</dc:creator>
<dc:creator>Jiang, Y.</dc:creator>
<dc:creator>Li, N.</dc:creator>
<dc:date>2025-07-16</dc:date>
<dc:identifier>doi:10.1101/2025.07.11.664267</dc:identifier>
<dc:title><![CDATA[A Paracrine-to-Autocrine Shunt of GREM1 Fuels Colorectal Cancer Metastasis via ACVR1C]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.07.16.665048v1?rss=1">
<title>
<![CDATA[
Barcoded Rabies In Situ Connectomics for high-throughput reconstruction of neural circuits 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.07.16.665048v1?rss=1"
</link>
<description><![CDATA[
Sequencing of oligonucleotide barcodes holds promise as a high-throughput approach for reconstructing synaptic connectivity at scale (1). Rabies viruses can act as a vehicle for barcode transmission, thanks to their ability to spread between synaptically connected cells (2, 3). However, applying barcoded rabies viruses to map synaptic connections in vivo has proved challenging (4-7). Here, we develop Barcoded Rabies In Situ Connectomics (BRISC) for high-throughput connectivity mapping in the mouse brain. To ensure that the majority of post-synaptic "starter" neurons are uniquely labeled with distinct barcode sequences, we first generated libraries of rabies viruses with sufficient diversity to label >1000 neurons uniquely. To minimize the probability of barcode transmission between starter neurons, we developed a strategy to tightly control their density. We then applied BRISC to map inputs of single neurons in the primary visual cortex (V1). Using in situ sequencing, we read out the expression of viral barcodes in rabies-infected neurons, while preserving spatial information. We then matched barcode sequences between starter and presynaptic neurons, mapping the inputs of 385 neurons and identifying 7,814 putative synaptic connections. The resulting connectivity matrix revealed layer- and cell-type-specific local connectivity rules and topographic organization of long-range inputs to V1. These results show that BRISC can simultaneously resolve the synaptic connectivity of hundreds of neurons while preserving spatial information, enabling reconstruction of neural circuits at an unprecedented scale.
]]></description>
<dc:creator>Becalick, A.</dc:creator>
<dc:creator>Blot, A.</dc:creator>
<dc:creator>Strom, M.</dc:creator>
<dc:creator>Znamenskiy, P.</dc:creator>
<dc:date>2025-07-20</dc:date>
<dc:identifier>doi:10.1101/2025.07.16.665048</dc:identifier>
<dc:title><![CDATA[Barcoded Rabies In Situ Connectomics for high-throughput reconstruction of neural circuits]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.07.21.665949v1?rss=1">
<title>
<![CDATA[
Boundary constraints can determine pattern emergence 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.07.21.665949v1?rss=1"
</link>
<description><![CDATA[
The robust patterning of cell fates during embryonic development requires precise coordination of signalling gradients within defined spatial constraints. Using a geometrically confined in vitro system derived from human embryonic stem cells, we demonstrate that patterning of neuromesodermal progenitors (NMPs) during axial elongation is driven by boundary-dependent mechanisms. Despite extensive work on radial fate patterning in confined 2D systems, the quantitative role of boundary conditions in shaping spatiotemporal dynamics remains unclear. Here, we show that a minimal reaction-diffusion model coupled with a simplified gene regulatory network accurately predicts spatial patterns across diverse geometries. Guided by its predictions, we identify Wnt signalling as a key component of the activator signal. Inhibition of Wnt secretion preserved initiation of patterning but disrupted subsequent morphogenesis, indicating distinct mechanisms govern pattern establishment versus maintenance. Our findings reveal how geometry encodes positional information that directs molecular patterning, providing insight into how spatial constraints and signalling dynamics guide robust tissue self-organisation during development.
]]></description>
<dc:creator>Loo, Y. T.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Harrison, R.</dc:creator>
<dc:creator>Rito, T.</dc:creator>
<dc:creator>Theis, S.</dc:creator>
<dc:creator>Charras, G.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Saunders, T. E.</dc:creator>
<dc:date>2025-07-22</dc:date>
<dc:identifier>doi:10.1101/2025.07.21.665949</dc:identifier>
<dc:title><![CDATA[Boundary constraints can determine pattern emergence]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.07.21.665861v1?rss=1">
<title>
<![CDATA[
Hippocampal circuit-specific enhancement of GABA-inhibition caused by discrete gene regions in a Down syndrome model 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.07.21.665861v1?rss=1"
</link>
<description><![CDATA[
Although Down syndrome (DS), trisomy 21, affects [~]6 million people worldwide, the neural circuit mechanisms underlying the neurophenotypes of impaired learning, memory and language are unknown. A prominent candidate mechanism involves dysfunctional GABA-signalling and GABAA receptor ligands have been proposed as therapeutics to reverse the neurophenotypic effects of DS.

By investigating GABA neurotransmission in brain regions important for cognition in mouse DS models, we reveal that excessive inhibition is not a ubiquitous feature of DS but instead is brain circuit-specific demonstrating increased phasic and tonic inhibition in the dentate gyrus with no comparative changes to inhibition in CA1 and medial prefrontal cortex.

In the dentate, elevated extrasynaptic GABA signalling, and interneuron numbers, likely underpin spike firing defects. We show that increased GABA inhibition is caused by increased dosage of Olig1, Olig2 and Dyrk1a. Overall, DS mice are characterised by circuit-specific dysfunctional inhibition predicted to affect cognition via sparse coding in the hippocampus.
]]></description>
<dc:creator>Hannan, S. B.</dc:creator>
<dc:creator>Lana-Elola, E.</dc:creator>
<dc:creator>Scales, S. W.</dc:creator>
<dc:creator>Fisher, E. M. C.</dc:creator>
<dc:creator>Tybulewicz, V. L. J.</dc:creator>
<dc:creator>Smart, T. G.</dc:creator>
<dc:date>2025-07-24</dc:date>
<dc:identifier>doi:10.1101/2025.07.21.665861</dc:identifier>
<dc:title><![CDATA[Hippocampal circuit-specific enhancement of GABA-inhibition caused by discrete gene regions in a Down syndrome model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.07.29.667237v1?rss=1">
<title>
<![CDATA[
Topographic alignment of auditory inputs to the visual cortex 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.07.29.667237v1?rss=1"
</link>
<description><![CDATA[
Sensory cortical areas send long-range projections to cortical areas from other sensory modalities, supporting multisensory integration to generate a unified representation of the external world. However, the organizational principles underlying these extensive cross-modal connections remain poorly understood. In this study, we investigated the anatomical and functional organisation of auditory cortex inputs in the visual cortex. We found that populations of anatomically segregated auditory cortex neurons project to different visual cortical areas, broadcasting distinct auditory information to the dorsal and ventral visual processing streams. While sound frequency information was homogenously distributed across visual cortical areas, sound location information was differentially broadcast across the visual cortex. Specifically, sound azimuth and elevation were differentially encoded across visual cortical areas and streams matching the retinotopic bias of the target area. These findings suggest that cross-modal cortico-cortical connections follow a simple rule whereby specialised projection pathways are topographically aligned with the organisational principles of the target sensory area, ensuring spatially coherent integration of multisensory signals.
]]></description>
<dc:creator>Egea-Weiss, A.</dc:creator>
<dc:creator>Turner-Bridger, B.</dc:creator>
<dc:creator>Viduolyte, A.</dc:creator>
<dc:creator>Marianelli, E.</dc:creator>
<dc:creator>Znamenskiy, P.</dc:creator>
<dc:creator>Iacaruso, M. F.</dc:creator>
<dc:date>2025-07-31</dc:date>
<dc:identifier>doi:10.1101/2025.07.29.667237</dc:identifier>
<dc:title><![CDATA[Topographic alignment of auditory inputs to the visual cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.07.29.667472v1?rss=1">
<title>
<![CDATA[
Multi-scale Molecular Imaging of Human Cells reveals COPI and COPII Vesicles at ER Exit Sites 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.07.29.667472v1?rss=1"
</link>
<description><![CDATA[
Trafficking from the endoplasmic reticulum to the Golgi apparatus comprises the first steps toward the correct localization of 30% of eukaryotic proteins. Coat protein complexes COPII and COPI are involved in forward and retrograde transport of cargo and cargo receptors between the ER and the Golgi. Although COPII forms coated vesicles in vitro, the biogenesis, morphology and organization of transport carriers in mammalian cells is debated. We use in situ cryo-electron tomography and super-resolution fluorescence microscopy to reveal the molecular architecture of ER exit sites in human cells. We visualise ribosome-exclusion zones enriched with COPII and COPI-coated vesicles and thus resolve the debate regarding the existence of COPII coated vesicles. COPII vesicles derive from ER membranes, whereas COPI vesicles originate from the ER-Golgi intermediate compartment. We quantify coated vesicle morphology and positioning with respect to other ER exit site components, providing a molecular description of the mammalian early secretory pathway.
]]></description>
<dc:creator>Downes, K. W.</dc:creator>
<dc:creator>Flood, J. R.</dc:creator>
<dc:creator>Nans, A.</dc:creator>
<dc:creator>Van der Verren, S.</dc:creator>
<dc:creator>Audhya, A.</dc:creator>
<dc:creator>Zanetti, G.</dc:creator>
<dc:date>2025-07-30</dc:date>
<dc:identifier>doi:10.1101/2025.07.29.667472</dc:identifier>
<dc:title><![CDATA[Multi-scale Molecular Imaging of Human Cells reveals COPI and COPII Vesicles at ER Exit Sites]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.08.04.668423v1?rss=1">
<title>
<![CDATA[
Integrative modeling of seasonal influenza evolution via AI-powered antigenic cartography 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.08.04.668423v1?rss=1"
</link>
<description><![CDATA[
Seasonal influenza viruses evade host immunity through rapid antigenic evolution. Antigenicity is assessed by serological assays and typically visualized as antigenic maps, which represent antigenic differences among virus strains. However, conventional maps cannot directly infer the antigenicity of unexamined variants from their genotypes. Here, we present PLANT, a protein language model that projects influenza A/H3N2 viruses onto an antigenic map using HA protein sequences. Using PLANT-based cartography, we show that (i) H3N2 antigenic evolution accelerates during periods of disrupted global circulation, (ii) antigenic novelty accounts for a substantial portion of viral fitness advantage, and (iii) vaccine strains are often antigenically distant from circulating viruses. We further propose a PLANT-based framework for selecting vaccine strains with improved antigenic match than the WHO-recommended strains. This study provides a statistical foundation for integrated modeling of viral genotype, antigenicity, and fitness, offering quantitative insights into seasonal influenza virus evolution and supporting rational vaccine design.
]]></description>
<dc:creator>Ito, J.</dc:creator>
<dc:creator>Kawakubo, S.</dc:creator>
<dc:creator>Unno, H.</dc:creator>
<dc:creator>Strange, A.</dc:creator>
<dc:creator>Lytras, S.</dc:creator>
<dc:creator>Okumura, K.</dc:creator>
<dc:creator>Lilley, A.</dc:creator>
<dc:creator>Harvey, R.</dc:creator>
<dc:creator>Lewis, N.</dc:creator>
<dc:creator>Sato, K.</dc:creator>
<dc:date>2025-08-04</dc:date>
<dc:identifier>doi:10.1101/2025.08.04.668423</dc:identifier>
<dc:title><![CDATA[Integrative modeling of seasonal influenza evolution via AI-powered antigenic cartography]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.08.06.668903v1?rss=1">
<title>
<![CDATA[
Extracting biological structure and heterogeneity from the nano to the macro scale 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.08.06.668903v1?rss=1"
</link>
<description><![CDATA[
Fluorescence microscopy is an essential tool in biology. It has revealed great variability at multiple scales, in macromolecular complexes, cells, and organisms. Understanding this variability will reveal the mechanisms by which genetically or biochemically identical systems adopt different biological states. Achieving this requires the ability to extract both the underlying biological structure and how it varies across the population. Currently the field lacks general techniques to deal with arbitrary structures and different types of variability. Here we present SQUASSH, a new convolutional neural network-based approach to freely fit structural models to fluorescence microscopy data that simultaneously quantifies variability to reveal correlations, dynamics, and systematic distortions. SQUASSH is highly versatile: it accommodates diverse imaging modalities at length scales from nm to mm. This approach opens up applications such as imaging nanoscale macromolecular structures, revealing patterns in shape changes from organelle to tissue scale, and characterizing systems biology of dynamical processes.
]]></description>
<dc:creator>Rosten, E.</dc:creator>
<dc:creator>Stedman, D.</dc:creator>
<dc:creator>Chu, L.-Y.</dc:creator>
<dc:creator>Wickramanayake, K.</dc:creator>
<dc:creator>Littlejohn, G.</dc:creator>
<dc:creator>Baxter, K. J.</dc:creator>
<dc:creator>McConnell, G.</dc:creator>
<dc:creator>Culley, S.</dc:creator>
<dc:creator>Ch'ng, Q.</dc:creator>
<dc:creator>Leterrier, C.</dc:creator>
<dc:creator>Bates, M.</dc:creator>
<dc:creator>Molodtsov, M.</dc:creator>
<dc:creator>Cox, S.</dc:creator>
<dc:date>2025-08-07</dc:date>
<dc:identifier>doi:10.1101/2025.08.06.668903</dc:identifier>
<dc:title><![CDATA[Extracting biological structure and heterogeneity from the nano to the macro scale]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.08.11.669575v1?rss=1">
<title>
<![CDATA[
Reconstructing Waddington's Landscape from Data 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.08.11.669575v1?rss=1"
</link>
<description><![CDATA[
The development of a zygote into a functional organism requires that this single progenitor cell gives rise to numerous distinct cell types. Attempts to exhaustively tabulate the interactions within developmental signaling networks that coordinate these hierarchical cell fate transitions are difficult to interpret or fit to data. An alternative approach models the cellular decision-making process as a flow in an abstract landscape whose signal-dependent topography defines the possible developmental outcomes and the transitions between them. Prior applications of this formalism have built landscapes in low-dimensional spaces without explicit reference to gene expression. Here, we present a computational geometry framework for fitting dynamical landscapes directly to high-dimensional single-cell data. Our method models the time evolution of probability distributions in gene expression space, enabling landscape construction with minimal free parameters and precise characterization of dynamical features, including fixed points, unstable manifolds, and basins of attraction. We demonstrate the applicability of this framework to multicolor flow-cytometry and RNA-seq data. Applied to a stem cell system that models ventral neural tube patterning, we recover a family of morphogen-dependent landscapes whose valleys align with canonical neural progenitor types. Remarkably, simple linear interpolation between landscapes captures signaling dependence, and chaining landscapes together reveals irreversible behavior following transient morphogen exposure. Our method combines the interpretability of landscape models with a direct connection to data, providing a general framework for understanding and controlling developmental dynamics.
]]></description>
<dc:creator>Cislo, D. J.</dc:creator>
<dc:creator>Delas, M. J.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Siggia, E. D.</dc:creator>
<dc:date>2025-08-13</dc:date>
<dc:identifier>doi:10.1101/2025.08.11.669575</dc:identifier>
<dc:title><![CDATA[Reconstructing Waddington's Landscape from Data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.08.08.669335v1?rss=1">
<title>
<![CDATA[
Inactivation of Airborne Pathogen Surrogates by Triethylene Glycol 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.08.08.669335v1?rss=1"
</link>
<description><![CDATA[
The COVID-19 outbreak brought to the fore the importance of airborne transmission in spreading human infectious diseases and highlighted the need for sustainable mitigation strategies. Triethylene glycol (TEG) has been documented as having microbicidal capabilities and has been proposed as one such mitigation strategy. Aerosolized TEG exhibits antimicrobial activity against airborne microorganisms; Grignard Pure Technology was developed to safely aerosolize TEG for decontamination of enclosed spaces. Here we show that this TEG formulation effectively inactivates airborne microorganisms, resulting in 2 to 4.5 net log reduction in concentration of viable bacteria, viruses, and mycobacteria within 30-60 minutes at TEG concentration (aerosol + vapor) of [~]0.7 mg/m3, which is well within the range considered safe for humans. Our data also demonstrate that aerosolizing both the test organisms and the antimicrobial product provides a more accurate and relevant measure of the products efficacy for indoor usage than traditional surface - or solution - based disinfection assays. Accurate evaluation of antimicrobial efficacy is a crucial step in adopting novel interventions and tools to control airborne pathogens that pose a public health risk. Our findings argue that testing protocols must match the intended use of any intervention. Given the safety concerns of aerosolizing human pathogens for direct testing of airborne infectious burden, we also advance an approach for selecting suitable surrogate microorganisms based on their phenotypic and biophysical similarity to corresponding pathogenic species.

ImportanceDuring the COVID-19 pandemic, personal protective equipment, social distancing, and even vaccinations proved sub-optimal in controlling the spread of COVID-19. Public health practice and the hierarchy of controls emphasize primary prevention, whereby the pathogen is removed or destroyed before exposure to the public. Triethylene glycol (TEG) has the potential to inactivate airborne pathogens and limit their spread. TEG is designated a "safer chemical" by the US EPA and has been used for decades in aerosol deodorizers and theatrical special effects. This study shows that aerosolized TEG is highly effective at eliminating a wide spectrum of viable airborne pathogen surrogates at concentrations well below the threshold of safety concern. Thus, it may afford significant protection against the transmission of infectious agents with pandemic potential.
]]></description>
<dc:creator>Desai, G.</dc:creator>
<dc:creator>Goldman, E.</dc:creator>
<dc:creator>Jordan, W.</dc:creator>
<dc:creator>Balarashti, J.</dc:creator>
<dc:creator>Caravanos, J.</dc:creator>
<dc:creator>Edgar, R. S.</dc:creator>
<dc:creator>Grignard, E.</dc:creator>
<dc:creator>Ramachandran, G.</dc:creator>
<dc:creator>Mainelis, G.</dc:creator>
<dc:date>2025-08-08</dc:date>
<dc:identifier>doi:10.1101/2025.08.08.669335</dc:identifier>
<dc:title><![CDATA[Inactivation of Airborne Pathogen Surrogates by Triethylene Glycol]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.08.20.671218v1?rss=1">
<title>
<![CDATA[
In situ cryo-electron tomography of vaccinia virus exit from infected cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.08.20.671218v1?rss=1"
</link>
<description><![CDATA[
Poxvirus-infected cells release newly assembled virions via Golgi-mediated envelopment and subsequent exocytosis at the plasma membrane, prior to cell lysis. Here, we used cryo-electron tomography and structured illumination microscopy to study vaccinia egress. Our 3D analysis reveals that Golgi-mediated envelopment is a flexible process that involves remodelling of the enfolding membrane and a final step that seals a small pore. During subsequent exocytosis, the viral outer membrane fuses with the plasma membrane but retains a distinct identity, beneath which septins and clathrin are independently recruited. Clathrin enhances actin-dependent viral spread, while septins suppress virus release from the cell. We found that septin filaments run parallel to the inner surface of the plasma membrane beneath virions attached to the cell surface. In contrast, clathrin induces the formation of plasma membrane invaginations in distinct subdomains. We propose that actin assembly at these subdomains provides a template for subsequent virus-induced actin polymerization.
]]></description>
<dc:creator>Hernandez-Gonzalez, M.</dc:creator>
<dc:creator>Nans, A.</dc:creator>
<dc:creator>Calcraft, T.</dc:creator>
<dc:creator>Rosenthal, P. B.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:date>2025-08-20</dc:date>
<dc:identifier>doi:10.1101/2025.08.20.671218</dc:identifier>
<dc:title><![CDATA[In situ cryo-electron tomography of vaccinia virus exit from infected cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.08.21.671502v1?rss=1">
<title>
<![CDATA[
Developmental changes in the carotid body transcriptome accompanying the maturation of chemosensitivity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.08.21.671502v1?rss=1"
</link>
<description><![CDATA[
The carotid body (CB) chemoreceptors mediate rapid cardiorespiratory reflexes to hypoxia, which mature peri-natally and are vital for fetal hypoxia tolerance and post-natal ventilatory control. This maturation is associated with an increase in the sensitivity of the CB electrophysiological response to hypoxia (chemosensitivity): a process that is incompletely understood but critical to systemic oxygen homeostasis. Hypothesizing that perinatal CB gene expression changes would reveal candidate mechanisms for oxygen chemosensitivity, we studied the CB transcriptome in sheep, where peri-natal CB physiology is well-characterised. CB-mediated cardiovascular reflexes are detectable at fetal day 120, and robust by term (day 145), while hypoxic ventilatory responses are established by post-natal day 15. We performed RNA sequencing on sheep CBs at each of these stages, and adults, along with the superior cervical ganglion (SCG) as an oxygen-insensitive control. This allowed us to define tissue-specific changes in the CB transcriptome correlating with chemosensitivity maturation. Striking, progressive CB enrichment is observed in genes implicated in murine CB chemosensitivity, including potassium channels (KCNK9), mitochondrial complex IV regulators (NDUFA4L2, HIGD1C), and HIF-2 (EPAS1). Genes with this expression pattern are also enriched for regulators of diacylglycerol (DAG), particularly the DAG kinase DGKH: one of the most abundant CB transcripts increasing in parallel with chemosensitivity. Across developmental stages, the CB also exhibits marked down-regulation of metabolic pathways and ATP/GTP consuming processes, potentially providing a state permissive to metabolic signal detection. Together, this builds a detailed picture of the CB transcriptional signature, with core features established in fetal life and conserved across species.

Key pointsO_LIThe carotid body (CB) chemoreceptors mediate rapid cardiorespiratory responses to hypoxia, which maintain systemic oxygen homeostasis, but CB dysfunction is also implicated in pathologies including hypertension, heart failure and sudden infant death.
C_LIO_LICB-mediated chemoreflexes mature during the peri-natal period, with increasing sensitivity of the oxygen chemosensory response.
C_LIO_LIWe performed RNA-seq of CBs from sheep across 3 peri-natal stages and adults, enabling us to identify gene expression changes that correlate with functional state.
C_LIO_LIWe describe a CB transcriptomic signature that is conserved across species, established in fetal life, and correlates with maturation. This includes features of a unique metabolic phenotype, and up-regulation of genes encoding the extracellular matrix and diacylglycerol signalling. The top transcription factor correlating with functional maturation is EPAS1/ HIF-2.
C_LIO_LIWe anticipate that this data set will be a valuable resource in generating novel hypotheses on mechanisms of oxygen chemosensory function, development and CB-associated pathologies.
C_LI
]]></description>
<dc:creator>Hodson, E. J.</dc:creator>
<dc:creator>Sugimoto, Y.</dc:creator>
<dc:creator>Manamperige, R. S.</dc:creator>
<dc:creator>Ford, S. G.</dc:creator>
<dc:creator>Botting-Lawford, K. J.</dc:creator>
<dc:creator>Niu, Y.</dc:creator>
<dc:creator>Nathan, J. A.</dc:creator>
<dc:creator>Giussani, D. A.</dc:creator>
<dc:creator>Ratcliffe, P. J.</dc:creator>
<dc:date>2025-08-25</dc:date>
<dc:identifier>doi:10.1101/2025.08.21.671502</dc:identifier>
<dc:title><![CDATA[Developmental changes in the carotid body transcriptome accompanying the maturation of chemosensitivity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.08.19.670819v1?rss=1">
<title>
<![CDATA[
α-Synuclein driven cell susceptibility in Parkinson's disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.08.19.670819v1?rss=1"
</link>
<description><![CDATA[
Early cellular events in Parkinsons disease (PD) remain elusive. While aggregation of -synuclein (Syn) into Lewy bodies marks advanced pathology, smaller Syn oligomers have been implicated in prodromal stages. Here we map Syn oligomers at single-particle resolution in post-mortem brain tissue from Braak stage 3/4 PD cases and matched controls. Quantitative imaging of 9,882 neurons across four regions captured over 112 million Syn oligomers. Mean intracellular -Syn burden was unchanged between groups, but PD samples contained a higher fraction of neurons whose oligomer load exceeded a specific aggregation threshold. We term these aggregation-susceptible cells (ASCs). ASC enrichment in vulnerable regions supports a population-level model in which early pathology arises from a stochastic shift in cellular composition rather than altered Syn aggregation kinetics. This human-tissue, large-scale dataset provides a quantitative framework for detecting ASCs and for testing population-level interventions in PD and related proteinopathies.
]]></description>
<dc:creator>Breiter, J. C.</dc:creator>
<dc:creator>Beckwith, J. S.</dc:creator>
<dc:creator>Brock, E. E.</dc:creator>
<dc:creator>Lachica, J.</dc:creator>
<dc:creator>Toomey, C. E.</dc:creator>
<dc:creator>Fu, B.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:creator>Weiss, L. E.</dc:creator>
<dc:creator>Wood, N. W.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Vendruscolo, M.</dc:creator>
<dc:creator>Lee, S. F.</dc:creator>
<dc:date>2025-08-23</dc:date>
<dc:identifier>doi:10.1101/2025.08.19.670819</dc:identifier>
<dc:title><![CDATA[α-Synuclein driven cell susceptibility in Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.08.26.672435v1?rss=1">
<title>
<![CDATA[
Balanced contractility and adhesion drive polarization in a minimal elastic actomyosin network 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.08.26.672435v1?rss=1"
</link>
<description><![CDATA[
Polarization of migrating cells involves chemical and mechanical interactions of signaling networks, cytoskeleton, plasma membrane, and substrate adhesions. Still, it is not fully understood which mechanisms and components are sufficient for symmetry breaking, and if they work independently or together. Here, we use a discrete active network model to investigate if and how an elastic cytoskeletal network is capable of breaking symmetry solely through mechanical interactions. Our minimal model consists of elastic bonds, attractive force dipoles, and force-sensitive anchor points, initially distributed uniformly and subject to simple turnover rules. We find that these features are sufficient to produce different cell behaviors, and, remarkably, to drive symmetry breaking and directed (polarized) motion. Network behavior was primarily determined by the turnover rate of anchor points, which, itself, is a function of the ratio between dipole force and the threshold force required for anchor removal. Directional motion emerged at intermediate turnover rates, at which tension in the network accumulated through several turnover cycles before eventually exceeding the adhesion removal threshold locally at the edge, mirroring our recent experimental findings on correlation of the traction force with protrusion-retraction transitions in the cell [1, 2]. At high turnover rates, forces were unable to build up to sufficiently high levels, while at low turnover rates, anchors hinder motion. These results demonstrate how directed motion can emerge as an intrinsic property of a simple mechanical network, independently of external cues or complex signaling networks. Given the concordance between this model and recent experimental findings, we suggest that polarization by contraction-adhesion dynamics could be a fundamental emergent behavior of actin-myosin networks.

Author summaryCells often need to move, for example, during development, wound healing, or immune responses. To do so, they must first decide where their "front" and "back" are, a process known as polarization. Most explanations for this behavior focus on complex chemical signaling inside the cell. In our work, we asked a simpler question: could mechanical forces within a cell be sufficient to make it polarize and move without an external cue? To explore this idea, we developed a computational model of a simplified cell made only of elastic connections, contractile forces, and attachment points to its surroundings. We started with a completely uniform system, without any built-in direction or external guidance. Surprisingly, we found that this minimal mechanical setup could spontaneously develop a front and a back and begin moving persistently. We proposed that the key mechanical factor controlling this behavior was the rate at which the attachment points break under mechanical load. When this process occurred at an intermediate rate, forces built up unevenly, leading to detachment and forward motion, similar to what is observed in-vitro. Our findings suggest that cell polarization and movement can emerge spontaneously from mechanical properties alone, highlighting an important and overlooked role for mechanics in cell behavior.
]]></description>
<dc:creator>Messi, Z.</dc:creator>
<dc:creator>Raynaud, F.</dc:creator>
<dc:creator>Goehring, N. W.</dc:creator>
<dc:creator>Verkhovsky, A. B.</dc:creator>
<dc:date>2025-08-29</dc:date>
<dc:identifier>doi:10.1101/2025.08.26.672435</dc:identifier>
<dc:title><![CDATA[Balanced contractility and adhesion drive polarization in a minimal elastic actomyosin network]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.04.29.591736v1?rss=1">
<title>
<![CDATA[
Multiomic analysis reveals developmental dynamics of the human heart in health and disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.04.29.591736v1?rss=1"
</link>
<description><![CDATA[
Developmental dynamics involve the specification of diverse cell types and their spatial organization into multicellular niches. Here, we combine single-cell and spatial multiomics to define 19 distinct tissue niches in the developing heart, leading to the development of a context-aware, resolution-agnostic niche classification tool (TissueTypist). Applying high-resolution spatial profiling to the developing sinoatrial node, we resolve three pacemaker cell subtypes arrayed along a linear axis. First trimester subpopulations, such as the pacemaker cells in the sinus horn and sinoatrial node head region, display neuro-attractant programmes and interact with parasympathetic neurons via interactions including Semaphorin-Plexin signalling. Temporal trajectories map maturation of atrial and ventricular cardiomyocytes, uncovering a lipid-metabolic switch and potential key regulators of cell type identity. In the ventricle, we identify cellular and transcriptional gradients along both pseudotime and transmural axes, offering new molecular insights into myocardial compaction and maturation. Comparative profiling of euploid and trisomy 21 hearts shows a depletion of compact cardiomyocytes and heightened apoptosis, validated in isogenic-matched trisomy 21 and euploid iPSC-derived cardiomyocytes. This implicates disrupted myocardial growth may be a mechanism for Downs syndrome-associated congenital heart disease. Overall, we deliver a spatially resolved framework of human cardiac development, enabling systematic exploration of developmental niches in health and disease.
]]></description>
<dc:creator>Cranley, J.</dc:creator>
<dc:creator>Kanemaru, K.</dc:creator>
<dc:creator>Bayraktar, S.</dc:creator>
<dc:creator>Knight-Schrijver, V.</dc:creator>
<dc:creator>Pett, J. P.</dc:creator>
<dc:creator>Polanski, K.</dc:creator>
<dc:creator>Dabrowska, M.</dc:creator>
<dc:creator>Mulas, I.</dc:creator>
<dc:creator>Richardson, L.</dc:creator>
<dc:creator>Semprich, C.</dc:creator>
<dc:creator>Kapuge, R.</dc:creator>
<dc:creator>Perera, S.</dc:creator>
<dc:creator>He, X.</dc:creator>
<dc:creator>Ho, S. Y.</dc:creator>
<dc:creator>Yayon, N.</dc:creator>
<dc:creator>Tuck, L.</dc:creator>
<dc:creator>Roberts, K.</dc:creator>
<dc:creator>Palmer, J.</dc:creator>
<dc:creator>Davaapil, H.</dc:creator>
<dc:creator>Gambardella, L.</dc:creator>
<dc:creator>Patel, M.</dc:creator>
<dc:creator>Tyser, R.</dc:creator>
<dc:creator>Sinha, S.</dc:creator>
<dc:creator>Teichmann, S.</dc:creator>
<dc:date>2024-04-29</dc:date>
<dc:identifier>doi:10.1101/2024.04.29.591736</dc:identifier>
<dc:title><![CDATA[Multiomic analysis reveals developmental dynamics of the human heart in health and disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.08.25.671921v1?rss=1">
<title>
<![CDATA[
Microglial activation and alpha-synuclein oligomers drive the early inflammatory phase of Parkinson's disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.08.25.671921v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is characterised by insoluble -synuclein (Syn) aggregates in Lewy bodies (LBs) within the substantia nigra, with cortical pathology appearing as the disease progresses. Late-stage LB deposition, cellular stress, and neuronal loss obscure disease-driving events, we therefore performed multi-regional transcriptomic and aggregate profiling in early-midstage PD brains (Braak 3-4), where cortical regions are pathologically unaffected. We report neuroimmune activation as an early PD feature, characterised by the expansion of a high-SNCA-expressing microglial state. This robust immune signature occurs prior to LB formation, but is associated with oligomeric Syn within cortical microglia. In hiPSC-derived microglia, both endogenous Syn oligomerisation, and exogenous oligomer uptake, trigger transcriptional reprogramming, characterised by interferon-driven inflammation, antigen presentation, and mitochondrial suppression, closely mirroring the early PD brain. These findings describe mechanisms by which Syn oligomerisation potently initiates early neuroinflammation, highlighting a critical interplay between proteinopathy and immune activation at the earliest stages of disease.
]]></description>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>Grant-Peters, M.</dc:creator>
<dc:creator>Beckwith, J. S.</dc:creator>
<dc:creator>Toomey, C. E.</dc:creator>
<dc:creator>Popovic, R.</dc:creator>
<dc:creator>Breiter, J. C.</dc:creator>
<dc:creator>Brenton, J. W.</dc:creator>
<dc:creator>Fairbrother-Browne, A.</dc:creator>
<dc:creator>Strohbuecker, S.</dc:creator>
<dc:creator>Lachica, J.</dc:creator>
<dc:creator>Rodriguez-Lopez, M.</dc:creator>
<dc:creator>Brock, E. E.</dc:creator>
<dc:creator>Fu, B.</dc:creator>
<dc:creator>Nahidiazar, L.</dc:creator>
<dc:creator>Lopez-Garcia, P.</dc:creator>
<dc:creator>Ferguson, R.</dc:creator>
<dc:creator>Saleeb, R. S.</dc:creator>
<dc:creator>Lucas-Clarke, H.</dc:creator>
<dc:creator>Penverne, A.</dc:creator>
<dc:creator>D'Sa, K.</dc:creator>
<dc:creator>Pang, C. W.</dc:creator>
<dc:creator>Horrocks, M. H.</dc:creator>
<dc:creator>Vendruscolo, M.</dc:creator>
<dc:creator>Wood, N.</dc:creator>
<dc:creator>Lee, S. F.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:date>2025-08-28</dc:date>
<dc:identifier>doi:10.1101/2025.08.25.671921</dc:identifier>
<dc:title><![CDATA[Microglial activation and alpha-synuclein oligomers drive the early inflammatory phase of Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.08.27.672654v1?rss=1">
<title>
<![CDATA[
Plasmodium ARK1 regulates spindle formation during atypical mitosis and forms a divergent chromosomal passenger complex 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.08.27.672654v1?rss=1"
</link>
<description><![CDATA[
Mitosis in Plasmodium spp., the causative agent of malaria, is fundamentally different from model eukaryotes, proceeding via a bipartite microtubule organising centre (MTOC) and lacking canonical regulators such as Polo kinases. During schizogony, asynchronous nuclear replication produces a multinucleate schizont, while rapid male gametogony generates an octaploid nucleus before gamete formation. Here, we identify Aurora-related kinase 1 (ARK1) as a key component of inner MTOC and spindle formation, controlling kinetochore dynamics and driving mitotic progression. Conditional ARK1 depletion disrupts spindle biogenesis, kinetochore segregation, karyokinesis and cytokinesis in both stages, and affects parasite transmission. Interactome analysis shows that ARK1 forms the catalytic core of a non-canonical chromosomal passenger complex (CPC) containing two highly divergent inner centromere proteins (INCENPs), which we term INCENP-A and INCENP-B, and lacking the canonical chromatin-targeting subunits Survivin and Borealin. Comparative genomics suggests that apicomplexan INCENPs arose through recurrent lineage-specific duplications, reflecting an evolutionary rewiring of CPC architecture in this eukaryotic lineage. Together, these findings reveal key adaptations in Plasmodium mitosis involving ARK1 and its INCENP scaffolds, and identify the ARK1-INCENP interface as a potential multistage target for antimalarial intervention.
]]></description>
<dc:creator>Nagar, A.</dc:creator>
<dc:creator>Yanase, R.</dc:creator>
<dc:creator>Zeeshan, M.</dc:creator>
<dc:creator>Ferguson, D. J. P.</dc:creator>
<dc:creator>Abel, S.</dc:creator>
<dc:creator>Pashley, S. L.</dc:creator>
<dc:creator>Mishra, A.</dc:creator>
<dc:creator>Eze, A.</dc:creator>
<dc:creator>Rea, E.</dc:creator>
<dc:creator>Brady, D.</dc:creator>
<dc:creator>Bottrill, A. R.</dc:creator>
<dc:creator>Vaughan, S.</dc:creator>
<dc:creator>Le Roch, K. G.</dc:creator>
<dc:creator>Guttery, D. S.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:creator>Tromer, E. C.</dc:creator>
<dc:creator>Sharma, P.</dc:creator>
<dc:creator>Tewari, R.</dc:creator>
<dc:date>2025-08-27</dc:date>
<dc:identifier>doi:10.1101/2025.08.27.672654</dc:identifier>
<dc:title><![CDATA[Plasmodium ARK1 regulates spindle formation during atypical mitosis and forms a divergent chromosomal passenger complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.02.673701v1?rss=1">
<title>
<![CDATA[
Critical point drying of brain tissue for X-ray phase contrast imaging 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.02.673701v1?rss=1"
</link>
<description><![CDATA[
X-ray phase contrast tomography is emerging as a powerful method for imaging large volumes of brain tissue at sub-cellular resolution. However, current sample preparation methods are largely inherited from visible light or electron microscopy workflows and hence are not optimised to exploit the full potential of X-ray contrast mechanisms. Here we propose to replace interstitial material by air to enhance X-ray phase contrast of the ultrastructural features. We used critical point drying (CPD) of heavy metal-stained mouse brain tissue to produce mechanically stable samples with preserved ultrastructure and enhanced refractive index boundaries, a nanofoam-like material that remains compatible with follow-up conventional resin embedding. Using two complementary synchrotron-based setups, a high-throughput microtomography beamline (P14, DESY) and a nanoscale holographic tomography beamline (ID16A, ESRF), we found that CPD samples consistently showed 2-4x stronger phase-shift signal than conventional resin-embedded tissue. The contrast gain remained consistent across samples, imaging conditions, and beamlines. Our results suggest that CPD offers a versatile route for preparing tissue for subcellular and ultrastructural-resolution X-ray imaging. It retains structural detail while improving signal, and is compatible with other processing procedures like femtosecond laser milling or electron microscopy, paving the path for biological tissue imaging beyond the mm3 scale.
]]></description>
<dc:creator>Khan, S.</dc:creator>
<dc:creator>Albers, J.</dc:creator>
<dc:creator>Vorobyev, A.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Reichmann, J.</dc:creator>
<dc:creator>Svetlove, A.</dc:creator>
<dc:creator>de Marco, F.</dc:creator>
<dc:creator>Denisova, K.</dc:creator>
<dc:creator>Yang, Y.</dc:creator>
<dc:creator>Seichepine, F.</dc:creator>
<dc:creator>Douglas, J. O.</dc:creator>
<dc:creator>Duke, E.</dc:creator>
<dc:creator>Cloetens, P.</dc:creator>
<dc:creator>Pacureanu, A.</dc:creator>
<dc:creator>Schaefer, A.</dc:creator>
<dc:creator>Bosch, C.</dc:creator>
<dc:date>2025-09-07</dc:date>
<dc:identifier>doi:10.1101/2025.09.02.673701</dc:identifier>
<dc:title><![CDATA[Critical point drying of brain tissue for X-ray phase contrast imaging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.06.674661v1?rss=1">
<title>
<![CDATA[
Near real-time data on the human neutralizing antibody landscape to influenza virus to inform vaccine-strain selection in September 2025 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.06.674661v1?rss=1"
</link>
<description><![CDATA[
The hemagglutinin of human influenza virus evolves rapidly to erode neutralizing antibody immunity. Twice per year, new vaccine strains are selected with the goal of providing maximum protection against the viruses that will be circulating when the vaccine is administered [~]8-12 months in the future. To help inform this selection, here we quantify how the antibodies in recently collected human sera neutralize viruses with hemagglutinins from contemporary influenza strains. Specifically, we use a high-throughput sequencing-based neutralization assay to measure how 188 human sera collected from Oct 2024 to April 2025 neutralize 140 viruses representative of the H3N2 and H1N1 strains circulating in humans as of the summer of 2025. This data set, which encompasses 26,148 neutralization titer measurements, provides a detailed portrait of the current human neutralizing antibody landscape to influenza A virus. The full data set and accompanying visualizations are available for use in vaccine development and viral forecasting.
]]></description>
<dc:creator>Kikawa, C.</dc:creator>
<dc:creator>Huddleston, J.</dc:creator>
<dc:creator>Loes, A. N.</dc:creator>
<dc:creator>Turner, S. A.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Barr, I. G.</dc:creator>
<dc:creator>Cowling, B. J.</dc:creator>
<dc:creator>Englund, J. A.</dc:creator>
<dc:creator>Greninger, A. L.</dc:creator>
<dc:creator>Harvey, R.</dc:creator>
<dc:creator>Hasegawa, H.</dc:creator>
<dc:creator>Ho, F.</dc:creator>
<dc:creator>Lacombe, K.</dc:creator>
<dc:creator>Leung, N. H.</dc:creator>
<dc:creator>Lewis, N. S.</dc:creator>
<dc:creator>Peck, H.</dc:creator>
<dc:creator>Watanabe, S.</dc:creator>
<dc:creator>Smith, D. J.</dc:creator>
<dc:creator>Bedford, T.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2025-09-06</dc:date>
<dc:identifier>doi:10.1101/2025.09.06.674661</dc:identifier>
<dc:title><![CDATA[Near real-time data on the human neutralizing antibody landscape to influenza virus to inform vaccine-strain selection in September 2025]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.08.30.673054v1?rss=1">
<title>
<![CDATA[
Lactate Blocks Tertiary Lymphoid Structure Formation by Inhibiting B Cell Chemotaxis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.08.30.673054v1?rss=1"
</link>
<description><![CDATA[
Tertiary lymphoid structures (TLS) and B cell infiltration are strong predictors of immunotherapy success across cancers, including triple-negative breast cancer (TNBC). However, immune-cold TNBCs often lack both features. Here, we identify a tumor-intrinsic mechanism that actively suppresses B cell recruitment. Despite evidence of B cell responses in cancer-associated lymph nodes (cLNs), B cells fail to infiltrate TNBC tumors or form TLS. This exclusion is not simply due to chemokine deficiency as exogenous chemokine addition fails to restore B cell migration. Using fractionation and metabolic profiling, we identify lactate as a dominant tumor-secreted metabolite that directly impairs B cell chemotaxis by disrupting mitochondrial metabolism. In vivo, combining lactate inhibition with engineered chemokine secretion promotes cLN-derived B cell infiltration and enables TLS formation, particularly when coupled with CD40 stimulation. Transcriptomics analyses across several human cancer datasets strengthen the association between high glycolytic activity with poor B-cell infiltration in chemokine-rich tumors. Together, our findings reveal lactate as a key metabolic barrier to B cell trafficking and TLS induction, suggesting that metabolic reprogramming may provide an avenue to convert "immune-cold" tumors into TLS-rich, immunologically responsive microenvironments.
]]></description>
<dc:creator>Calado, D. P.</dc:creator>
<dc:creator>Boulat, V.</dc:creator>
<dc:creator>Alberts, E.</dc:creator>
<dc:creator>Driscoll, P. C.</dc:creator>
<dc:creator>Hung, M. S.</dc:creator>
<dc:creator>Cunha, A.</dc:creator>
<dc:creator>Quist, J.</dc:creator>
<dc:creator>Liu, F.</dc:creator>
<dc:creator>Brundin, C. A.</dc:creator>
<dc:creator>Bhalla, A.</dc:creator>
<dc:creator>Avalle, L.</dc:creator>
<dc:creator>Rosekilly, J.</dc:creator>
<dc:creator>Ryan, L.</dc:creator>
<dc:creator>Gillett, C.</dc:creator>
<dc:creator>Strom, M.</dc:creator>
<dc:creator>Poli, V.</dc:creator>
<dc:creator>MacRae, J. I.</dc:creator>
<dc:creator>Grigoriadis, A.</dc:creator>
<dc:date>2025-09-04</dc:date>
<dc:identifier>doi:10.1101/2025.08.30.673054</dc:identifier>
<dc:title><![CDATA[Lactate Blocks Tertiary Lymphoid Structure Formation by Inhibiting B Cell Chemotaxis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.11.675585v1?rss=1">
<title>
<![CDATA[
Streptococcus pneumoniae accessory capsular genes modulate fitness, pathogenicity and immune evasion 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.11.675585v1?rss=1"
</link>
<description><![CDATA[
Globally, Streptococcus pneumoniae disproportionally affects children in resource-poor settings, older adults and people living with HIV. Frequently found as an asymptomatic colonizer of the nasopharynx, this versatile pathogen is a prominent cause of pneumonia, meningitis, bacteraemia and otitis media. Recently, a serotype 3 capsule variant (GPSC10-ST700) has expanded in Malawi with enhanced vaccine escape potential. Here, using a mutational and complementation approach, we show that loss of accessory capsular genes in GPSC10-ST700 contribute to increased opsonophagocytic resistance in this lineage. Although originally thought to be nonfunctional pseudogenes, we show that these genes modulate fitness and the global phosphoproteome in serotype 3 strains. These findings highlight that vaccine escape may be mediated through variations in the pneumococcal capsular locus that enhance fitness, pathogenicity and immune evasion, without capsule switching.

IMPORTANCEPneumococcal polysaccharide-conjugate vaccines (PCV) target the polysaccharide capsule (CPS), which is a dominant virulence factor. However, current PCVs induce suboptimal protection against serotype 3 strains, which produce a thicker capsule that when released from the bacterial surface, interferes with antibody-mediated bacterial killing and protection. We recently described the clonal expansion of a sequence type (ST) 700-GPSC10 serotype 3 lineage in Malawi post-PCV13 introduction. This lineage is characterized by the absence of at least 6 genes in its cps locus and a distinct antimicrobial resistance (AMR) profile compared to other serotype 3 strains. Here we uncovered a functional role for the accessory capsular genes (acl) in serotype 3, previously considered to be pseudogenes, which modulate capsule production, shedding, serum tolerance, and bacterial fitness. By linking genotype to phenotype, our work provides new insights into the molecular basis of serotype 3 immune evasion, informing the design of more effective pneumococcal vaccines.
]]></description>
<dc:creator>Chan, J. M.</dc:creator>
<dc:creator>Flynn, H. R.</dc:creator>
<dc:creator>Kim, S.</dc:creator>
<dc:creator>Demetriou, H.</dc:creator>
<dc:creator>Ramos-Sevillano, E.</dc:creator>
<dc:creator>Alsari, A.</dc:creator>
<dc:creator>Roalfe, L.</dc:creator>
<dc:creator>Ercoli, G.</dc:creator>
<dc:creator>Kalizang'oma, A.</dc:creator>
<dc:creator>Brown, C.</dc:creator>
<dc:creator>Chaguza, C.</dc:creator>
<dc:creator>French, N.</dc:creator>
<dc:creator>Brown, J. S.</dc:creator>
<dc:creator>Goldblatt, D.</dc:creator>
<dc:creator>de Carvalho, L. P. S.</dc:creator>
<dc:creator>Heyderman, R.</dc:creator>
<dc:date>2025-09-15</dc:date>
<dc:identifier>doi:10.1101/2025.09.11.675585</dc:identifier>
<dc:title><![CDATA[Streptococcus pneumoniae accessory capsular genes modulate fitness, pathogenicity and immune evasion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.15.676266v1?rss=1">
<title>
<![CDATA[
Investigations into fission yeast chromosome size determinants 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.15.676266v1?rss=1"
</link>
<description><![CDATA[
Mitotic chromosome dimensions differ between organisms, and they differ within a species between developmental stages. The physiological determinants of chromosome size remain poorly understood. Here, we investigate chromosome size determinants in the fission yeast Schizosaccharomyces pombe. Super-resolution microscopy and semi-automated measurements reveal that cell or nuclear volume in interphase, or the time spent in mitosis (both previously proposed chromosome size determinants), have little influence on resultant chromosome dimensions. Instead, levels of the chromosomal condensin complex affect chromosome size, with increasing condensin levels resulting in more compact, shorter and thinner, chromosomes. These observations inform our understanding of how chromosome dimensions are controlled in an organism. They suggest that a chromosome-intrinsic mechanism sets chromosome size, more so than the environment in which chromosomes find themselves in.
]]></description>
<dc:creator>Wu, P.-S.</dc:creator>
<dc:creator>Fallesen, T.</dc:creator>
<dc:creator>Uhlmann, F.</dc:creator>
<dc:date>2025-09-15</dc:date>
<dc:identifier>doi:10.1101/2025.09.15.676266</dc:identifier>
<dc:title><![CDATA[Investigations into fission yeast chromosome size determinants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.12.675937v1?rss=1">
<title>
<![CDATA[
Immunogenicity and Efficacy of Digitally Immune Optimised H1N1 Vaccine Candidates in Swine and Murine Animal Models 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.12.675937v1?rss=1"
</link>
<description><![CDATA[
Influenza A virus (IAV) zoonotic transmission and constant evolution in multiple species heightens the risk of emerging novel strains at the human-animal interface. Composite antigens including hemagglutinin (HA), neuraminidase (NA), and matrix-2 (M2) proteins were computationally designed to maximize the breadth of the immune response elicited to human seasonal, pandemic, and zoonotic H1N1 IAVs. Mouse hyperimmune serum raised against these antigens demonstrated broad H1 neutralization and N1 inhibition activity. To enhance immunogenicity, the antigens were combined as a single DNA expression construct (DVX-H1N1). Studies in the well-recognized swine model for human influenza demonstrated that DVX-H1N1 immunization induced broad, neutralizing antibody responses and markedly reduced nasal shedding of viral RNA following challenge with 1A.3.3.2 subclade strain A/swine/England/1353/2009 (H1N1). An effective immune response and reduction in virus shedding was observed in pigs immunized with a whole inactivated virus (WIV) vaccine homologous to the challenge strain but not with a human-origin seasonal WIV vaccine. Overall, we demonstrated broad immunogenicity and efficacy of the DVX-H1N1 vaccine candidate, benchmarked against relevant IAV H1N1 strains in vitro and in vivo in mice and pigs.

IMPORTANCEThe zoonotic potential of swine-origin IAVs is a recognized global health threat. Vaccination remains the most effective intervention against influenza; protecting at the population level by preventing nasal shedding and transmission, but also in individuals by limiting clinical disease, particularly by reducing the severity of lung infection. The World Health Organization (WHO) spearheads biannual surveillance efforts to review evolving virus strains and vaccine antigens at Vaccine Candidate Meetings (VCM) to recommend strain updates for the human seasonal influenza vaccine and for pandemic preparedness purposes. However, the strain selection approach is complex and efficaciousness of seasonal influenza vaccines still varies significantly based on the accurate matching of the predicted strains in circulation with the manufactured vaccine antigens. This emphasizes the need for next-generation influenza vaccines that improve the breadth and longevity of immunity. We describe a computationally optimized DNA vaccine with broad immunogenicity and robust efficacy in the pig model.
]]></description>
<dc:creator>Del Rosario, J. M. M.</dc:creator>
<dc:creator>van Diemen, P. M.</dc:creator>
<dc:creator>Frost, S.</dc:creator>
<dc:creator>vishwanath, s.</dc:creator>
<dc:creator>Sujit, S. B.</dc:creator>
<dc:creator>Ashokan, S. K.</dc:creator>
<dc:creator>Mollett, B. C.</dc:creator>
<dc:creator>Ramsay, A. M.</dc:creator>
<dc:creator>Simpson, B.</dc:creator>
<dc:creator>Asbach, B.</dc:creator>
<dc:creator>Carnell, G. W.</dc:creator>
<dc:creator>Chan, A.</dc:creator>
<dc:creator>Tonks, P.</dc:creator>
<dc:creator>Temperton, N. J.</dc:creator>
<dc:creator>Davies, M.</dc:creator>
<dc:creator>McCauley, J. W.</dc:creator>
<dc:creator>Kinsley, R.</dc:creator>
<dc:creator>Wagner, R.</dc:creator>
<dc:creator>Everett, H. E.</dc:creator>
<dc:creator>Heeney, J. L.</dc:creator>
<dc:date>2025-09-14</dc:date>
<dc:identifier>doi:10.1101/2025.09.12.675937</dc:identifier>
<dc:title><![CDATA[Immunogenicity and Efficacy of Digitally Immune Optimised H1N1 Vaccine Candidates in Swine and Murine Animal Models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.10.675313v1?rss=1">
<title>
<![CDATA[
SLFN11 restricts escape from telomere crisis to prevent alternative lengthening of telomeres 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.10.675313v1?rss=1"
</link>
<description><![CDATA[
The tRNA nuclease SLFN11 is epigenetically silenced in [~]50% of treatment-naive tumours and is the strongest predictor of chemoresistance but why it is frequently inactivated in cancer is unknown. To acquire immortality, cancer cells can activate alternative lengthening of telomeres (ALT), typically accompanied by ATRX loss. Here, we implicate SLFN11 in sensing telomere replication stress, triggering eradication of ATRX deficient cells prior to ALT establishment. Whereas progressive telomere shortening of cells lacking telomerase and ATRX leads to telomere crisis and cell death, SLFN11 loss confers tolerance to PML-BLM dependent ALT intermediates, permitting emergence of ALT survivors. We propose that during tumorigenesis SLFN11 inactivation is selected as means to tolerate endogenous replication stress following telomere crisis, leading to the development of therapy resistant tumours before treatment.
]]></description>
<dc:creator>Segura-Bayona, S.</dc:creator>
<dc:creator>Borel, V.</dc:creator>
<dc:creator>Stanage, T. H.</dc:creator>
<dc:creator>Maric, M.</dc:creator>
<dc:creator>Walter, M.</dc:creator>
<dc:creator>Oliveira, R. A.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Idilli, A. I.</dc:creator>
<dc:creator>Peritore, M.</dc:creator>
<dc:creator>Hewitt, G.</dc:creator>
<dc:creator>Hekkelman, M.</dc:creator>
<dc:creator>Snell, D. M.</dc:creator>
<dc:creator>Jones, S. T.</dc:creator>
<dc:creator>Cheshire, C.</dc:creator>
<dc:creator>Elezi, A.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Mitter, R.</dc:creator>
<dc:creator>Mikolajczak, A.</dc:creator>
<dc:creator>Gee, H. E.</dc:creator>
<dc:creator>Nye, E.</dc:creator>
<dc:creator>O'Sullivan, R. J.</dc:creator>
<dc:creator>Singhi, A. D.</dc:creator>
<dc:creator>Brummelkamp, T. R.</dc:creator>
<dc:creator>Cesare, A. J.</dc:creator>
<dc:creator>Boulton, S. J.</dc:creator>
<dc:date>2025-09-10</dc:date>
<dc:identifier>doi:10.1101/2025.09.10.675313</dc:identifier>
<dc:title><![CDATA[SLFN11 restricts escape from telomere crisis to prevent alternative lengthening of telomeres]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.04.673943v1?rss=1">
<title>
<![CDATA[
TDP-43 loss of function drives aberrant splicing in Parkinson's disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.04.673943v1?rss=1"
</link>
<description><![CDATA[
Introductory paragraphWhile mRNA splicing dysregulation is a well-established contributor to neurodegeneration in disorders such as amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), its role in Parkinsons disease (PD) remains underexplored. Here, we analyse transcriptomic data from >500 post-mortem human brain samples from individuals with and without PD to show that splicing alterations are frequently detected. Differentially spliced genes were significantly more enriched for those causally-implicated in both PD and ALS than genes that were differentially expressed. Furthermore, we observed a strong association between these splicing alterations and dysfunction of the RNA-binding protein (RBP), TAR DNA-binding protein 43 (TDP-43). Strikingly, genes and exon junctions affected by TDP-43 knockdown overlapped significantly with those dysregulated across brain regions in PD. In brains from individuals with the LRRK2 c.6055G>A (p.G2019S) mutation, the most common genetic cause of PD, we also observed significant enrichment of TDP-43-dependent splicing changes. This finding was corroborated in human pluripotent stem cell-derived midbrain dopaminergic neurons and a LRRK2 p.G2019S knock-in mouse model, where reduced nuclear TDP-43 levels evidenced the well-recognised loss-of-function mechanism contributing to splicing dysregulation. By leveraging our RNA-based analyses we predicted TDP-43-dependent novel peptide sequences and validated their existence within human LRRK2 mutation mDNs, while also demonstrating an overall loss of protein and mRNA expression in mis-spliced genes. Collectively, our findings reveal that PD is marked by extensive splicing dysregulation dependent on TDP-43, making TDP-43 a promising new therapeutic target in PD.
]]></description>
<dc:creator>Brenton, J. W.</dc:creator>
<dc:creator>Follett, J.</dc:creator>
<dc:creator>Nirujogi, R.</dc:creator>
<dc:creator>Toomey, C. E.</dc:creator>
<dc:creator>Lopez-Garcia, P.</dc:creator>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>Lee, Y. J.</dc:creator>
<dc:creator>Syed, K. M.</dc:creator>
<dc:creator>Rocamora Perez, G.</dc:creator>
<dc:creator>Fairbrother-Browne, A.</dc:creator>
<dc:creator>D'Sa, K.</dc:creator>
<dc:creator>Grant-Peters, M.</dc:creator>
<dc:creator>Lachica, J.</dc:creator>
<dc:creator>Hicks, A. R.</dc:creator>
<dc:creator>Wagen, A. Z.</dc:creator>
<dc:creator>O'Callaghan, B.</dc:creator>
<dc:creator>Macpherson, H.</dc:creator>
<dc:creator>Montgomery, K.-a.</dc:creator>
<dc:creator>Busquets, O.</dc:creator>
<dc:creator>Reynolds, R. H.</dc:creator>
<dc:creator>Garcia Ruiz, S.</dc:creator>
<dc:creator>Cao, T.</dc:creator>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Plun-Favreau, H.</dc:creator>
<dc:creator>Wong, P. C.</dc:creator>
<dc:creator>Farrer, M.</dc:creator>
<dc:creator>Lashley, T.</dc:creator>
<dc:creator>Soldner, F.</dc:creator>
<dc:creator>Hockemeyer, D.</dc:creator>
<dc:creator>Alessi, D.</dc:creator>
<dc:creator>Wood, N. W.</dc:creator>
<dc:creator>Hardy, J.</dc:creator>
<dc:creator>Rio, D. C.</dc:creator>
<dc:creator>Jaunmuktane, Z.</dc:creator>
<dc:creator>Gustavsson, E. K.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:date>2025-09-09</dc:date>
<dc:identifier>doi:10.1101/2025.09.04.673943</dc:identifier>
<dc:title><![CDATA[TDP-43 loss of function drives aberrant splicing in Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.18.677011v1?rss=1">
<title>
<![CDATA[
Chaperone AIP Couples mTORC1 Activation and Catabolic Metabolism During Neonatal Development 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.18.677011v1?rss=1"
</link>
<description><![CDATA[
To grow and divide cells must tightly coordinate anabolic programs with the availability of nutrients and growth factors. This balance is especially critical during postnatal development, when biosynthetic and energetic demands are high, and nutrient supply and neonates have to adapt to periods of fasting. These conditions place acute stress on the proteostasis network, making autophagy essential for nutrient recycling. We found that the chaperone aryl hydrocarbon receptor-interacting protein (AIP) supports both arms of this metabolic balance: promoting anabolic PI3K-AKT signaling for mTORC1 activation and enabling catabolic processes such as proteasomal degradation and autophagy. Loss of AIP causes a severe neonatal metabolic disorder, where affected infants fail to thrive postnatally. Our findings establish AIP as a central regulator of neonatal metabolic adaptation and cellular homeostasis.

One Sentence SummaryAIP integrates nutrient sensing and protein recycling to sustain neonatal survival.
]]></description>
<dc:creator>Korbonits, M.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Barry, S.</dc:creator>
<dc:creator>Lim, C. T.</dc:creator>
<dc:creator>Suleyman, O.</dc:creator>
<dc:creator>De Tito, S.</dc:creator>
<dc:creator>Uddin, N. B.</dc:creator>
<dc:creator>Vignola, M. L.</dc:creator>
<dc:creator>Hall, C.</dc:creator>
<dc:creator>Perna, L.</dc:creator>
<dc:creator>Chapple, P.</dc:creator>
<dc:creator>Henson, S.</dc:creator>
<dc:creator>Morales, V.</dc:creator>
<dc:creator>Bianchi, K.</dc:creator>
<dc:creator>Edvardsson, V. O.</dc:creator>
<dc:creator>Ragnarsson, K. A.</dc:creator>
<dc:creator>Kristinsdottir, V. E.</dc:creator>
<dc:creator>Debeer, A.</dc:creator>
<dc:creator>Sleyp, Y.</dc:creator>
<dc:creator>Zinchenko, R. A.</dc:creator>
<dc:creator>Anderson, G.</dc:creator>
<dc:creator>Duchen, M.</dc:creator>
<dc:creator>Singh, K.</dc:creator>
<dc:creator>Chung, C.-Y.</dc:creator>
<dc:creator>Yuan, Y.</dc:creator>
<dc:creator>Patel, S.</dc:creator>
<dc:creator>Aksoy, E.</dc:creator>
<dc:creator>Borovikov, A.</dc:creator>
<dc:creator>Bjornsson, H. T.</dc:creator>
<dc:creator>Van Esch, H.</dc:creator>
<dc:creator>Czibik, G.</dc:creator>
<dc:creator>Tooze, S.</dc:creator>
<dc:creator>Brennan, C. H.</dc:creator>
<dc:creator>Haworth, O.</dc:creator>
<dc:date>2025-09-18</dc:date>
<dc:identifier>doi:10.1101/2025.09.18.677011</dc:identifier>
<dc:title><![CDATA[Chaperone AIP Couples mTORC1 Activation and Catabolic Metabolism During Neonatal Development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.17.674891v1?rss=1">
<title>
<![CDATA[
Mechanisms for cross-neutralisation of diverse bat sarbecoviruses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.17.674891v1?rss=1"
</link>
<description><![CDATA[
The continuing evolution of SARS-CoV-2 variants of concern, and the increasing spillover potential of sarbecoviruses into the human population presents an important and urgent need to discover cross-reactive monoclonal antibodies (mAbs) for future therapeutic use and identify conserved neutralising epitopes that can be used for rationale design of broadly protective sarbecovirus vaccines. Here we study the neutralising epitopes on WIV-1 Spike of three mAbs that confer broad sarbecovirues and SARS-CoV-2 variant neutralisation, including XEC and JN.1. mAb V1WT_06 binds a highly conserved RBD site V epitope that is mediated by the heavy chain alone. V1WT_06 contact residues are highly conserved in circulating viruses suggesting that the epitope is evolutionarily and functionally constrained. mAbs V1WT_41 and VA14_26 bind overlapping RBD class 4 epitopes with differing angles of approach that impact on the degree of ACE2 competition. We show that neutralisation by these mAbs is maintained when virus entry is via Japanese horseshoe bat and Halcyon horseshoe bat ACE2. These mAbs are ideal candidates for therapeutic antibody development and inform the rational design of pan-coronavirus vaccines.
]]></description>
<dc:creator>Upadhyay, A.</dc:creator>
<dc:creator>Seow, J.</dc:creator>
<dc:creator>Alguel, Y.</dc:creator>
<dc:creator>Newman, J.</dc:creator>
<dc:creator>Thakur, N.</dc:creator>
<dc:creator>Hay, A.</dc:creator>
<dc:creator>Tam, J. C. H.</dc:creator>
<dc:creator>Nans, A.</dc:creator>
<dc:creator>Orton, R.</dc:creator>
<dc:creator>Bailey, D.</dc:creator>
<dc:creator>Cherepanov, P.</dc:creator>
<dc:creator>Doores, K. J.</dc:creator>
<dc:date>2025-09-17</dc:date>
<dc:identifier>doi:10.1101/2025.09.17.674891</dc:identifier>
<dc:title><![CDATA[Mechanisms for cross-neutralisation of diverse bat sarbecoviruses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.15.676088v1?rss=1">
<title>
<![CDATA[
Morphogen and juxtacrine signalling dynamically integrate to specify cell fates with single-cell resolution 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.15.676088v1?rss=1"
</link>
<description><![CDATA[
Morphogen gradients guide tissue patterning but do not act in isolation. How they integrate with other signalling modalities, like juxtacrine signalling, and how these integrations influence pattern resolution and robustness remain unclear. We address this in the Drosophila lamina, where columns of precursors are patterned with single-cell resolution into motion-processing neurons, dependent on a photoreceptor-derived Hedgehog gradient and glial-orchestrated differentiation. Combining experiments and theory, we show that glial-induced ERK activity drives Delta expression in lamina precursors, generating a graded Notch activity pattern. Notch restricts Hedgehog morphogen relay and enhances positional information. Glia act as timekeepers, scheduling ERK-driven differentiation after Hedgehog and Notch patterns are established. Thus, Notch and ERK dynamically integrate with Hedgehog to encode positional information, enabling reproducible cell fate patterning with single-cell resolution.
]]></description>
<dc:creator>Donoghue, A.</dc:creator>
<dc:creator>Mosby, L.</dc:creator>
<dc:creator>Lago-Baldaia, I.</dc:creator>
<dc:creator>Hodgetts, T.</dc:creator>
<dc:creator>Ursu, E.</dc:creator>
<dc:creator>Erten, Z.</dc:creator>
<dc:creator>Hadjivasiliou, Z.</dc:creator>
<dc:creator>Fernandes, V. M.</dc:creator>
<dc:date>2025-09-17</dc:date>
<dc:identifier>doi:10.1101/2025.09.15.676088</dc:identifier>
<dc:title><![CDATA[Morphogen and juxtacrine signalling dynamically integrate to specify cell fates with single-cell resolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.17.674754v1?rss=1">
<title>
<![CDATA[
Conserved Aberrant Developmental Trajectories of Human and Mouse SBMA Motor Neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.17.674754v1?rss=1"
</link>
<description><![CDATA[
Spinal bulbar muscular atrophy (SBMA) is a neuromuscular disease caused by a polyglutamine repeat expansion in the androgen receptor gene (AR). Lower motor neuron loss is a key feature of the disease, yet it remains poorly understood why these cells are affected. The transcriptional mechanisms underlying SBMA pathogenesis and how these evolve across developmental and disease stages remains incompletely defined.

To elucidate the molecular mechanisms underlying motor neuron loss in SBMA, we first performed transcriptomic profiling of both induced pluripotent stem cell derived motor neurons (iPSC-MNs) generated from SBMA patients and laser-captured micro dissected motor neurons (LCM-MNs) from symptomatic AR100 SBMA mice. We compared differential gene expression between the two models to identify shared transcriptional programs. To address the temporal progression of molecular changes we conducted profiling at key stages of motor neurogenesis in the developing iPSC-MNs and at pre-symptomatic and end-stage disease in AR100 SBMA mice to elucidate the emergence of the transcriptional phenotype and the trajectory of the gene expression changes.

We found significant transcriptional convergence between these two species. Notably, shared dysregulation was observed in pathways related to the spliceosome, the cell cycle and mitochondrial function. These transcriptional alterations emerged early in motor neurogenesis suggesting a developmental component to SBMA. Further in AR100 LCM-MNs we also observed disruption of mitochondrial and DNA damage repair pathways from pre-symptomatic to end stage disease.

This study identifies conserved pathogenic mechanisms across two SBMA model systems and provides crucial insights into the molecular basis and temporal dynamics of SBMA progression which may help identify potential therapeutic targets for SBMA.



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=110 SRC="FIGDIR/small/674754v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Devine, H.</dc:creator>
<dc:creator>Roberts, M. J.</dc:creator>
<dc:creator>Ziff, O. J.</dc:creator>
<dc:creator>Hanna, M. G.</dc:creator>
<dc:creator>Greensmith, L.</dc:creator>
<dc:creator>Patani, R.</dc:creator>
<dc:creator>Malik, B.</dc:creator>
<dc:date>2025-09-17</dc:date>
<dc:identifier>doi:10.1101/2025.09.17.674754</dc:identifier>
<dc:title><![CDATA[Conserved Aberrant Developmental Trajectories of Human and Mouse SBMA Motor Neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.17.676809v1?rss=1">
<title>
<![CDATA[
Characterisation of LAMP1- and LAMP2A-positive organelles in neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.17.676809v1?rss=1"
</link>
<description><![CDATA[
LAMP1 and LAMP2A are abundant proteins of late endosomal/lysosomal compartments, which are often used interchangeably to label what is thought to be the same pool of organelles, potentially obscuring their unique physiological roles. Here, we characterised the transport dynamics of LAMP1- and LAMP2A-positive compartments in human iPSC-derived cortical neurons. We found that axonal LAMP1-positive organelles move more slowly in the retrograde direction, pause more frequently, and show a broader velocity distribution in the anterograde direction than LAMP2A-positive vesicles, suggesting they are distinct compartments with differential trafficking behaviour. To explore the molecular mechanism underlying these differences, we characterised with high spatiotemporal precision, the protein interactomes of LAMP1 and LAMP2A-positive compartments through proximity labelling, using full-length LAMP1 or LAMP2A fused to the light-activated biotin ligase LOV-Turbo. We identified and validated the endosomal protein, ZFYVE16, as a novel member of LAMP1 and LAMP2A interactomes. We suggest that LAMP2A-positive organelles represent a subset of LAMP1-positive compartments, which are surprisingly enriched in synaptic vesicle proteins.

Summary statementLAMP1- and LAMP2A-positive organelles have different axonal transport dynamics and form distinct organelle pools characterised by specific protein compositions.
]]></description>
<dc:creator>Abouward, R.</dc:creator>
<dc:creator>Abdelhafid, A. M.</dc:creator>
<dc:creator>Wilkins, O. G.</dc:creator>
<dc:creator>Lee, S.-Y.</dc:creator>
<dc:creator>Ibrahim, F.</dc:creator>
<dc:creator>Skehel, M.</dc:creator>
<dc:creator>Ting, A.</dc:creator>
<dc:creator>Birsa, N.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Schiavo, G.</dc:creator>
<dc:date>2025-09-17</dc:date>
<dc:identifier>doi:10.1101/2025.09.17.676809</dc:identifier>
<dc:title><![CDATA[Characterisation of LAMP1- and LAMP2A-positive organelles in neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.15.675999v1?rss=1">
<title>
<![CDATA[
Cell migration sculpts evolutionary dynamics favouring therapy resistance in lung cancer 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.15.675999v1?rss=1"
</link>
<description><![CDATA[
The evolution of cancer undermines the long-term efficacy of therapy. In this study, we combine experimental, computational, and clinical data analysis to investigate factors influencing the competition of subclones in lung cancer. Lineage tracing reveals unexpected variation in the long-term fate of neutral subclones, with subclones arising near the edge of tumours being favoured. Low levels of cell migration and cell mixing lead to high cell densities in the interior of the tumour that suppress proliferation. Using agent-based modelling and in silico analysis we inferred the extent of cell mixing in human tumours from the TRACERx lung cancer study. This reveals correlations between Epithelial to Mesenchymal Transition (EMT), stromal fibroblasts and levels of cell mixing. Experimental analysis confirms that both TGF{beta}-driven EMT and stromal fibroblasts reduce the variability in subclone fate and promote subclone mixing. Moreover, mixing favours clonal sweeps by subclones resistant to therapy-induced cell killing. Together, these analyses demonstrate that EMT and stromal fibroblasts sculpt tumour evolution by promoting cell mixing and thereby favour the rapid dominance of therapy resistant subclones.
]]></description>
<dc:creator>Bhargava, A.</dc:creator>
<dc:creator>Fu, X.</dc:creator>
<dc:creator>Bailey, S.</dc:creator>
<dc:creator>Naito, Y.</dc:creator>
<dc:creator>Mohammadi, H.</dc:creator>
<dc:creator>Hynds, R. E.</dc:creator>
<dc:creator>Hooper, S.</dc:creator>
<dc:creator>Biswas, D.</dc:creator>
<dc:creator>Le Marois, A.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Alexandrov, Y.</dc:creator>
<dc:creator>French, P. M.</dc:creator>
<dc:creator>McGinty, J.</dc:creator>
<dc:creator>Bates, P. A.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Sahai, E.</dc:creator>
<dc:date>2025-09-16</dc:date>
<dc:identifier>doi:10.1101/2025.09.15.675999</dc:identifier>
<dc:title><![CDATA[Cell migration sculpts evolutionary dynamics favouring therapy resistance in lung cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.22.677761v1?rss=1">
<title>
<![CDATA[
Distinct ATRX functions cooperate with 9-1-1 and CST complexesto safeguard replication and telomere integrity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.22.677761v1?rss=1"
</link>
<description><![CDATA[
Mutations in the ATRX chromatin remodeller predispose to a developmental genetic disorder and cancer, but how it safeguards genome and telomere stability remains unresolved. Here, we uncover critical dependencies for the CTC1-STN1-TEN1 (CST) complex and RAD9A-HUS1-RAD1 (9-1-1) clamp in ATRX deficient cells. ATRX:CST synthetic lethality manifests following accumulation of telomeric G-rich ssDNA, which results in telomere loss and cell death. Conversely, we attribute ATRX:9-1-1 synthetic lethality to genome-wide ssDNA lesions, which compromise DNA replication. We further show ATRX suppresses DNA damage during replication stress by counteracting the activity of the FAM111A protease. We demonstrate that roles of ATRX in telomere maintenance and replication are genetically separable requiring its ATPase activity and PIP-box, respectively, and independently of its DAXX interaction. Collectively, functions of ATRX in suppressing toxic ssDNA lesions are context-dependent and are key to global DNA replication and telomere integrity.
]]></description>
<dc:creator>Segura-Bayona, S.</dc:creator>
<dc:creator>Maric, M.</dc:creator>
<dc:creator>Takaki, T.</dc:creator>
<dc:creator>Manova, Z.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Stanage, T. H.</dc:creator>
<dc:creator>Idilli, A. I.</dc:creator>
<dc:creator>Hewitt, G.</dc:creator>
<dc:creator>Ruis, P.</dc:creator>
<dc:creator>Millar, R.</dc:creator>
<dc:creator>Patel, H.</dc:creator>
<dc:creator>Howell, S.</dc:creator>
<dc:creator>Kotsantis, P.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>Boulton, S. J.</dc:creator>
<dc:date>2025-09-22</dc:date>
<dc:identifier>doi:10.1101/2025.09.22.677761</dc:identifier>
<dc:title><![CDATA[Distinct ATRX functions cooperate with 9-1-1 and CST complexesto safeguard replication and telomere integrity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.18.677100v1?rss=1">
<title>
<![CDATA[
Systems-level feedback loops maintain gene expression homeostasis following RNA polymerase II dosage perturbation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.18.677100v1?rss=1"
</link>
<description><![CDATA[
Transcription is regulated by sequence-specific transcription factors and enzymes allowing access to genes in chromatin. However, recent data indicate that the abundance of RNA polymerase II (RNAPII) itself may under certain circumstances represent an additional, crucial determinant of transcription regulation. Here, we used the dTAG system to titrate the cellular dosage of human RPB1, the largest RNAPII subunit, to more generally assess the importance of RNAPII levels. Unexpectedly, cells are extremely sensitive to RPB1 dosage, with a mere 30% reduction sufficient to perturb cell proliferation, cell cycle progression, and global transcription. Importantly, alterations in RPB1 abundance trigger hierarchical gene expression changes that are highly organized rather than stochastic. Using a combination of sequencing and proteomic approaches, we uncover the existence of multiple feedback loops between transcriptional initiation, promotor-proximal pause release, transcript elongation, splicing, and mRNA half-life, which together establish RNAPII abundance as a crucial systems-level regulator of transcriptional homeostasis.
]]></description>
<dc:creator>Olthof, A. M.</dc:creator>
<dc:creator>Gonzalez, M. N.</dc:creator>
<dc:creator>Poulsen, N. B.</dc:creator>
<dc:creator>Moreno, N. N.</dc:creator>
<dc:creator>Bautu, T.</dc:creator>
<dc:creator>Cugusi, S.</dc:creator>
<dc:creator>Svejstrup, J.</dc:creator>
<dc:date>2025-09-18</dc:date>
<dc:identifier>doi:10.1101/2025.09.18.677100</dc:identifier>
<dc:title><![CDATA[Systems-level feedback loops maintain gene expression homeostasis following RNA polymerase II dosage perturbation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.24.678353v1?rss=1">
<title>
<![CDATA[
Peptide molecular glues select between BET paralogues by exploiting allosteric sites and conformational dynamics 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.24.678353v1?rss=1"
</link>
<description><![CDATA[
Achieving selective target inhibition is critical for minimising drug side effects. This can be especially challenging when targeting individual members of protein families with high sequence similarity. A well-recognised example is the Bromodomain and Extraterminal domain (BET) family of proteins. Chemical inhibition of the acetyllysine (AcK)-binding bromodomains (BDs) of BET proteins has shown considerable promise in a range of disease models. However, despite over a decade of medicinal chemistry efforts, it has proven challenging to develop BET BD inhibitors that exhibit high selectivity between BET family paralogues. Cyclic peptides are versatile scaffolds for therapeutic development and often exhibit high selectivity and affinity for their targets. To explore their potential as selective BET BD inhibitors, we have used mRNA display to identify cyclic peptide ligands for the N-terminal BD of BRD2 and BRD4. The structurally diverse cyclic peptides enriched from the selections boast superior selectivity and affinity to previously developed inhibitors. Most strikingly, we isolated cyclic peptides with [~]1000-fold higher affinity for their target BD over the paralogous BDs, far surpassing selectivities reported to date. Our biochemical and structural data suggest that paralogue-selective cyclic peptides act as molecular glues, exploiting both subtle sequence differences at locations far from the AcK-binding pocket and differences in conformational dynamics between BET BD paralogues to achieve this unprecedented level of specificity. This work provides a blueprint for the development of new classes of selective BET inhibitors and, more generally, underscores the potential of exploiting protein dynamics in the design of selective ligands.
]]></description>
<dc:creator>Patel, K.</dc:creator>
<dc:creator>Mohanty, B.</dc:creator>
<dc:creator>Norman, A.</dc:creator>
<dc:creator>Franck, C.</dc:creator>
<dc:creator>Pachl, P.</dc:creator>
<dc:creator>Yang, L.</dc:creator>
<dc:creator>Jing, X.</dc:creator>
<dc:creator>Reid, X. J.</dc:creator>
<dc:creator>Walshe, J. L.</dc:creator>
<dc:creator>Solomon, P.</dc:creator>
<dc:creator>Tran, D. H.</dc:creator>
<dc:creator>Ford, D.</dc:creator>
<dc:creator>Low, J. K. K.</dc:creator>
<dc:creator>Wilkinson-White, L.</dc:creator>
<dc:creator>Passioura, T.</dc:creator>
<dc:creator>Payne, R. J.</dc:creator>
<dc:creator>Suga, H.</dc:creator>
<dc:creator>Walport, L. J.</dc:creator>
<dc:creator>Mackay, J. P.</dc:creator>
<dc:date>2025-09-24</dc:date>
<dc:identifier>doi:10.1101/2025.09.24.678353</dc:identifier>
<dc:title><![CDATA[Peptide molecular glues select between BET paralogues by exploiting allosteric sites and conformational dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.26.678648v1?rss=1">
<title>
<![CDATA[
Combinatorial protein barcodes enable self-correcting neuron tracing with nanoscale molecular context 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.26.678648v1?rss=1"
</link>
<description><![CDATA[
Mapping nanoscale neuronal morphology with molecular annotations is critical for understanding healthy and dysfunctional brain circuits. Current methods are constrained by image segmentation errors and by sample defects (e.g., signal gaps, section loss). Genetic strategies promise to overcome these challenges by using easily distinguishable cell identity labels. However, multicolor approaches are spectrally limited in diversity, whereas nucleic acid barcoding lacks a cellfilling morphology signal for segmentation. Here, we introduce PRISM (Protein-barcode Reconstruction via Iterative Staining with Molecular annotations), a platform that integrates combinatorial delivery of antigenically distinct, cell-filling proteins with tissue expansion, multi-cycle imaging, barcode-augmented reconstruction, and molecular annotation. Protein barcodes increase label diversity by >750-fold over multicolor labeling and enable morphology reconstruction with intrinsic error correction. We acquired a [~]10 million {micro}m3 volume of mouse hippocampal area CA2/3, multiplexed across 23 barcode antigen and synaptic marker channels. By combining barcodes with shape information, we achieve an 8x increase in automatic tracing accuracy of genetically labelled neurons. We demonstrate PRISM supports automatic proofreading across micron-scale spatial gaps and reconnects neurites across discontinuities spanning hundreds of microns. Using PRISMs molecular annotation capability, we map the distribution of synapses onto traced neural morphology, characterizing challenging synaptic structures such as thorny excrescences (TEs), and discovering a size correlation among spatially proximal TEs on the same dendrite. PRISM thus supports selfcorrecting neuron reconstruction with molecular context.
]]></description>
<dc:creator>Park, S. Y.</dc:creator>
<dc:creator>Sheridan, A.</dc:creator>
<dc:creator>An, B.</dc:creator>
<dc:creator>Jarvis, E.</dc:creator>
<dc:creator>Lyudchik, J.</dc:creator>
<dc:creator>Patton, W.</dc:creator>
<dc:creator>Axup, J. Y.</dc:creator>
<dc:creator>Chan, S. W.</dc:creator>
<dc:creator>Damstra, H. G. J.</dc:creator>
<dc:creator>Leible, D.</dc:creator>
<dc:creator>Leung, K. S.</dc:creator>
<dc:creator>Magno, C. A.</dc:creator>
<dc:creator>Meeran, A.</dc:creator>
<dc:creator>Michalska, J. M.</dc:creator>
<dc:creator>Rieger, F.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Wu, M.</dc:creator>
<dc:creator>Church, G. M.</dc:creator>
<dc:creator>Funke, J.</dc:creator>
<dc:creator>Huffman, T.</dc:creator>
<dc:creator>Leeper, K. G. C.</dc:creator>
<dc:creator>Truckenbrodt, S.</dc:creator>
<dc:creator>Winnubst, J.</dc:creator>
<dc:creator>Kornfeld, J. M. R.</dc:creator>
<dc:creator>Boyden, E. S.</dc:creator>
<dc:creator>Rodriques, S. G.</dc:creator>
<dc:creator>Payne, A. C.</dc:creator>
<dc:date>2025-09-28</dc:date>
<dc:identifier>doi:10.1101/2025.09.26.678648</dc:identifier>
<dc:title><![CDATA[Combinatorial protein barcodes enable self-correcting neuron tracing with nanoscale molecular context]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.24.678208v1?rss=1">
<title>
<![CDATA[
CLONAL SELECTION SUPPORTED BY SINGLE CELL DNA SEQUENCING REVEALS HORMONAL ADAPTATION AND RESISTANCE IN LOCALLY ADVANCED BREAST CANCER DURING NEOADJUVANT AROMATASE INHIBITION 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.24.678208v1?rss=1"
</link>
<description><![CDATA[
BackgroundThe aromatase inhibitors (AI) letrozole and exemestane, are often used in sequence in targeting ER+ breast cancers. However resistance to AI poses a major barrier to sustained clinical benefit, while the biological mechanisms underlying the phenomenon remain largely unknown. In this study, we build on our clinical NeoLetExe trial, with the aim to investigate the molecular basis of resistance to AI, by analysing subclonal evolutionary dynamics during sequential treatment.

MethodsWe use whole-exome sequencing (WES) data from 11 ER+ breast cancer patients and 3 timepoints of the Neoletexe trial to reconstruct cancer cell fraction-based subclonal composition. Single-cell DNA sequencing from matched tumour samples is used for validating identified clones and variants. Subclonal variants were annotated to genes by integrating evidence from public data and ExpectoSc. Pathway enrichment analysis using Human Base was conducted.

ResultsHigher cancer cell fraction clone trajectories were significantly associated with reduced treatment response (p = 0.023). Clones reconstructed by WES were validated at 81% using single-cell DNA sequencing. Clones resistant to both letrozole and exemestane demonstrated PIK3CA/AKT/mTOR signaling activation, KRAS pathway dysregulation, hedgehog signaling, and androgen receptor pathways, alongside extensive immune activation and metabolic reprogramming. Drug-specific resistance patterns showed exemestane-resistant clones enriched for epigenetic control and miRNA-mediated silencing, while letrozole-resistant clones displayed metabolic dysregulation but notably lacked immune pathway activation. In contrast, treatment-sensitive clones maintained coordinated cell cycle control, preserved DNA damage responses, and retained immune signaling capacity. Analysis of FDA-approved breast cancer targets identified actionable alterations in PIK3CA (4 patients) and AKT1 (1 patient) that persisted through AI treatment, with RNA expression analysis revealing 48 additional therapeutic targets spanning PI3K/AKT/mTOR, CDK4/6, DNA repair (BRCA1/2, ATM), and immune checkpoint pathways.

ConclusionWES-based cancer cell fraction analysis successfully captured subclonal evolutionary trajectories during AI treatment, revealing drug-specific mechanisms and identifying key molecular players in endocrine therapy resistance. This work establishes a framework for precision oncology approaches by providing actionable therapeutic targets and advancing our understanding of resistance mechanisms to improve clinical outcomes in sequential AI therapy.
]]></description>
<dc:creator>O'Mahony, D. G.</dc:creator>
<dc:creator>Lesluyes, T.</dc:creator>
<dc:creator>Brorson, I. S.</dc:creator>
<dc:creator>Vernhoff, P.</dc:creator>
<dc:creator>Sokolova, K.</dc:creator>
<dc:creator>Aure, M. R.</dc:creator>
<dc:creator>Alnaes, G. G.</dc:creator>
<dc:creator>Hooen, R. M.</dc:creator>
<dc:creator>Sundaram, A. Y.</dc:creator>
<dc:creator>Theesfeld, C.</dc:creator>
<dc:creator>Geisler, S. B.</dc:creator>
<dc:creator>Sauer, T.</dc:creator>
<dc:creator>Bahrami, N.</dc:creator>
<dc:creator>Parulkar, R.</dc:creator>
<dc:creator>Tahiri, A.</dc:creator>
<dc:creator>Luders, T.</dc:creator>
<dc:creator>Troyanskaya, O.</dc:creator>
<dc:creator>Vaske, C.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>Geisler, J.</dc:creator>
<dc:creator>Kristensen, V. N.</dc:creator>
<dc:date>2025-09-26</dc:date>
<dc:identifier>doi:10.1101/2025.09.24.678208</dc:identifier>
<dc:title><![CDATA[CLONAL SELECTION SUPPORTED BY SINGLE CELL DNA SEQUENCING REVEALS HORMONAL ADAPTATION AND RESISTANCE IN LOCALLY ADVANCED BREAST CANCER DURING NEOADJUVANT AROMATASE INHIBITION]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.30.679035v1?rss=1">
<title>
<![CDATA[
Splenic granulopoiesis and S100A9 drive resistance to checkpoint inhibitors conferred by liver metastases 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.30.679035v1?rss=1"
</link>
<description><![CDATA[
Here, we investigate why liver metastases reduce the efficacy of immune checkpoint inhibitors (CPI). The poor prognosis of patients with liver metastases is associated with a systemic increase in neutrophils. Using experimental models, we confirm that mice with liver metastases respond poorly to CPI, have elevated neutrophils and suppress the response of subcutaneous lesions to CPI. We demonstrate that liver metastases, acting partly via IL-6, boost granulopoiesis in the spleen and promote the generation of immature S100A9hi neutrophils that suppress T-cell proliferation. Human liver metastases exhibit a similar increase in S100A9hi neutrophils. Neutrophil depletion attenuates the growth of liver metastases, but not subcutaneous metastases. Moreover, genetic deletion of S100A9 enables liver metastases to be effectively treated with CPI, and prevents liver metastases from suppressing the response in subcutaneous metastases. Thus, we document how liver metastases specifically change splenic granulopoiesis leading to changes in the microenvironment of non-hepatic lesions, and how targeting a key neutrophil protein restores the efficacy of CPI.
]]></description>
<dc:creator>Lee, R.</dc:creator>
<dc:creator>Hooper, S.</dc:creator>
<dc:creator>Pallett, L.</dc:creator>
<dc:creator>Diniz, M.</dc:creator>
<dc:creator>Mckinnon-Snell, T.</dc:creator>
<dc:creator>Ramsden, Z.</dc:creator>
<dc:creator>Rabas, N.</dc:creator>
<dc:creator>Boeing, S.</dc:creator>
<dc:creator>Kennedy, O.</dc:creator>
<dc:creator>Buttercase, C.</dc:creator>
<dc:creator>Aidoo-Micah, G.</dc:creator>
<dc:creator>Price, G.</dc:creator>
<dc:creator>Macfarlane, S. C.</dc:creator>
<dc:creator>Bailey, S.</dc:creator>
<dc:creator>Davies, J.</dc:creator>
<dc:creator>Mathur, A.</dc:creator>
<dc:creator>Pistocchi, G.</dc:creator>
<dc:creator>Carminati, A.</dc:creator>
<dc:creator>Gupta, A.</dc:creator>
<dc:creator>Serra, P.</dc:creator>
<dc:creator>Davidson, B. R.</dc:creator>
<dc:creator>Pollok, J.-M.</dc:creator>
<dc:creator>Papayannopoulos, V.</dc:creator>
<dc:creator>Malanchi, I.</dc:creator>
<dc:creator>Lorigan, P.</dc:creator>
<dc:creator>Maini, M. K.</dc:creator>
<dc:creator>Sahai, E.</dc:creator>
<dc:date>2025-10-01</dc:date>
<dc:identifier>doi:10.1101/2025.09.30.679035</dc:identifier>
<dc:title><![CDATA[Splenic granulopoiesis and S100A9 drive resistance to checkpoint inhibitors conferred by liver metastases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.06.677012v1?rss=1">
<title>
<![CDATA[
Endoplasmic Reticulum Geometry Dictates Neuronal Bursting via Calcium Store Refill Rates 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.06.677012v1?rss=1"
</link>
<description><![CDATA[
The endoplasmic reticulum (ER)s continuous morphology is tightly controlled by ER-shaping proteins, whose genetic or expression defects drive a spectrum of neurodegenerative disorders from Hereditary Spastic Paraplegia to Alzheimers disease. Why perturbations in ER morphology manifest specifically in neurons remains unknown.

Here, by coupling visualisation of global sub-Hz firing bursts to ER ultrastructural manipulations in hiPSC-derived cortical neurons, alongside physical simulations, we establish a key ER structure-function principle: neuronal ER architecture dictates Ca2+ replenishment speed. Altering ER structure hinders network ER luminal connectivity and Ca2+ propagation from refill points at plasma membrane contact sites, impairing the ERs capability to supply repetitive Ca2+ bursts. The ER morpho-regulatory control of Ca2+ refill speed thus constitutes a switch on neuronal activity. These results expose the selective vulnerability of Ca2+-firing cells to ER structural disruptions, rationalising ER dysfunction in neurodegeneration. This mechanism could apply universally to Ca2+-firing cells.
]]></description>
<dc:creator>Davi, V.</dc:creator>
<dc:creator>Parutto, P.</dc:creator>
<dc:creator>Crapart, C.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Konno, T.</dc:creator>
<dc:creator>Chambers, J.</dc:creator>
<dc:creator>Franklin, J. P.</dc:creator>
<dc:creator>Maddison, D.</dc:creator>
<dc:creator>Awadelkareem, M. A.</dc:creator>
<dc:creator>Devine, M. J.</dc:creator>
<dc:creator>Koslover, E.</dc:creator>
<dc:creator>Avezov, E.</dc:creator>
<dc:date>2025-10-07</dc:date>
<dc:identifier>doi:10.1101/2025.10.06.677012</dc:identifier>
<dc:title><![CDATA[Endoplasmic Reticulum Geometry Dictates Neuronal Bursting via Calcium Store Refill Rates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.01.678762v1?rss=1">
<title>
<![CDATA[
Apicortin defines the Plasmodium apical conoid body during transmission but is dispensable for the parasite life cycle 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.01.678762v1?rss=1"
</link>
<description><![CDATA[
Apicomplexan parasites such as Plasmodium spp. and Toxoplasma gondii possess unique tubulin-based structures, including subpellicular microtubules and apical polar rings, which are essential for parasite motility, host cell invasion, and replication. How the stability of these structures is maintained is poorly understood, but it may involve Apicortin, a microtubule-associated protein, so-far found only in apicomplexans and the placozoan Trichoplax adhaerens. Apicortin contains a doublecortin (DC) domain and a partial tubulin polymerisation-promoting protein (TPPP) domain, both implicated in microtubule binding and stabilization. In this study, we investigated the location and function of Apicortin in Plasmodium berghei. Live cell imaging of a transgenic parasite line expressing GFP-tagged Apicortin showed that it was present at the apical end of invasive parasites only during development of transmission stages within the mosquito vector. High-resolution imaging using super-resolution and expansion microscopy, revealed that Apicortin forms a distinct ring-like structure in the apical complex region at the apical end of ookinetes and sporozoites. However, deletion of the Apicortin gene had no effect on parasite development and transmission through the mosquito, indicating that this protein is not essential. This suggests that there may be redundancy or compensatory functions in the mechanisms that stabilise the apical complex.
]]></description>
<dc:creator>Zeeshan, M.</dc:creator>
<dc:creator>Mishra, A.</dc:creator>
<dc:creator>Pashley, S. L.</dc:creator>
<dc:creator>Markus, R.</dc:creator>
<dc:creator>Brady, D.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:creator>Moores, C.</dc:creator>
<dc:creator>Tewari, R.</dc:creator>
<dc:date>2025-10-03</dc:date>
<dc:identifier>doi:10.1101/2025.10.01.678762</dc:identifier>
<dc:title><![CDATA[Apicortin defines the Plasmodium apical conoid body during transmission but is dispensable for the parasite life cycle]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.07.680949v1?rss=1">
<title>
<![CDATA[
Rapid expansion of podoplanin-positive fibroblasts following radiation limits the anti-tumour CD8+ T-cell response to radiotherapy. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.07.680949v1?rss=1"
</link>
<description><![CDATA[
Radiotherapy is known to cause changes in the tumour stroma which can undermine treatment efficacy. Our understanding of this process has historically centred around effects driven by Transforming Growth Factor-beta (TGF-{beta}) and alpha-smooth muscle actin (-SMA)+ fibroblasts. Here, we identified a rapid expansion of podoplanin (PDPN)+ fibroblasts following radiotherapy in breast, head and neck and melanoma tumours. This fibrosis was not dependent on TGF-{beta}, but was downstream of a radiotherapy-induced adaptive immune response. CD8+ T-cells entering the tumour after radiation were sequestered at the interface between residual tumour cells and PDPN+ fibroblasts and failed to enter the tumour core. Genetic deletion of PDPN in fibroblasts impacted their cytoskeleton and ability to organise extracellular matrix. This was associated with increased CD8+ T-cell entry and spontaneous tumour regression. Overall, we identify a mechanism whereby PDPN+ fibrosis limits immune-mediated radiation cell kill and demonstrate that disruption of PDPN signalling favours tumour control.

SignificanceIn this study we show that rapid podoplanin (PDPN)+ fibroblast expansion following radiotherapy limits immune-mediated radiation cell kill. Targeting PDPN and associated downstream signalling improves tumour control and is a promising strategy in combination with radiotherapy.

Graphical abstract

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]]></description>
<dc:creator>Wilkins, A. C.</dc:creator>
<dc:creator>Makris, S.</dc:creator>
<dc:creator>Green, M.</dc:creator>
<dc:creator>Hooper, S.</dc:creator>
<dc:creator>Mikolajczak, A.</dc:creator>
<dc:creator>Giangreco, G.</dc:creator>
<dc:creator>Allanki, S.</dc:creator>
<dc:creator>Nye, E.</dc:creator>
<dc:creator>Connick, E.</dc:creator>
<dc:creator>Mitter, R.</dc:creator>
<dc:creator>Cooper, S.</dc:creator>
<dc:creator>Durie, E.</dc:creator>
<dc:creator>Pedersen, M.</dc:creator>
<dc:creator>Burley, A.</dc:creator>
<dc:creator>O'Fee, K.</dc:creator>
<dc:creator>O'Leary, B.</dc:creator>
<dc:creator>Shewring, D.</dc:creator>
<dc:creator>Ramsden, Z.</dc:creator>
<dc:creator>Melcher, A.</dc:creator>
<dc:creator>Harrington, K.</dc:creator>
<dc:creator>Somaiah, N.</dc:creator>
<dc:creator>Acton, S. E.</dc:creator>
<dc:creator>Sahai, E.</dc:creator>
<dc:date>2025-10-07</dc:date>
<dc:identifier>doi:10.1101/2025.10.07.680949</dc:identifier>
<dc:title><![CDATA[Rapid expansion of podoplanin-positive fibroblasts following radiation limits the anti-tumour CD8+ T-cell response to radiotherapy.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.08.679225v1?rss=1">
<title>
<![CDATA[
DNMT3A-R882 mutations intrinsically drive dysfunctional neutropoiesis from human haematopoietic stem cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.08.679225v1?rss=1"
</link>
<description><![CDATA[
Clonal haematopoiesis (CH) arises from the expansion of hematopoietic stem cells (HSCs) carrying leukaemia-associated somatic mutations. CH is linked to pathological immune dysregulation and a greater risk of age-related inflammatory diseases. Yet, how CH mutations impact HSC differentiation into immune effector cells remains understudied. Here, we report a single-cell resolution functional and multi-omic investigation of HSC clonal and differentiation dynamics in individuals with DNMT3A-R882 CH. DNMT3A-R882 reshapes the clonal architecture of haematopoiesis towards an aged phylogenetic structure. Functionally, DNMT3A-R882 HSCs produce decreased monocytic output but more abundant and mature neutrophil progeny compared to WT HSCs in the same individual. Whereas DNMT3A-R882 myeloid progenitors display attenuated inflammatory transcriptional programmes, DNMT3A- R882 mature neutrophils acquire proinflammatory and immunomodulatory features typical of maladaptive immunity and CH co-morbidities.

Our findings, validated in humanised mice, identify aberrant DNMT3A-R882 HSC-driven neutropoiesis as a key link between CH, immune dysregulation and risk of inflammatory disease.
]]></description>
<dc:creator>Mantica, G.</dc:creator>
<dc:creator>Vedi, A.</dc:creator>
<dc:creator>Tuval, A.</dc:creator>
<dc:creator>Huerga Encabo, H.</dc:creator>
<dc:creator>Hayler, D.</dc:creator>
<dc:creator>Krzywon, A.</dc:creator>
<dc:creator>Mitchell, E.</dc:creator>
<dc:creator>Dunn, W. G.</dc:creator>
<dc:creator>Biezuner, T.</dc:creator>
<dc:creator>Bastos, H. P.</dc:creator>
<dc:creator>Sham, K.</dc:creator>
<dc:creator>Santoro, A.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Williams, N.</dc:creator>
<dc:creator>Danin, A.</dc:creator>
<dc:creator>Chapal, N.</dc:creator>
<dc:creator>Moskovitz, Y.</dc:creator>
<dc:creator>Arruda, A.</dc:creator>
<dc:creator>Fiorillo, E.</dc:creator>
<dc:creator>Orru, V.</dc:creator>
<dc:creator>Marongiu, M.</dc:creator>
<dc:creator>McKinney, E.</dc:creator>
<dc:creator>Cucca, F.</dc:creator>
<dc:creator>Collin, M.</dc:creator>
<dc:creator>Minden, M.</dc:creator>
<dc:creator>Campbell, P.</dc:creator>
<dc:creator>Vassiliou, G. S.</dc:creator>
<dc:creator>Fabre, M. A.</dc:creator>
<dc:creator>Nangalia, J.</dc:creator>
<dc:creator>Bonnet, D.</dc:creator>
<dc:creator>Shlush, L.</dc:creator>
<dc:creator>Laurenti, E.</dc:creator>
<dc:date>2025-10-08</dc:date>
<dc:identifier>doi:10.1101/2025.10.08.679225</dc:identifier>
<dc:title><![CDATA[DNMT3A-R882 mutations intrinsically drive dysfunctional neutropoiesis from human haematopoietic stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.08.681035v1?rss=1">
<title>
<![CDATA[
Continuous evolution of a halogenase enzyme with improved solubility and activity for sustainable bioproduction 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.08.681035v1?rss=1"
</link>
<description><![CDATA[
Halogenation enhances the stability and function of pharmaceuticals, biomaterials, and industrial compounds. However, chemical halogenation lacks stereoselectivity and requires the use of toxic or expensive chemicals. Although enzymatic halogenation can improve selectivity and reduce environmental impact, current halogenases are inefficient and insoluble, leading to low yields that limit their applications. Here, we develop RebHEvo4, a soluble and highly active tryptophan halogenase, containing 12 mutations that confer 37-fold and 44-fold increases in 7-chloro and 7-bromotryptophan production respectively, in vivo. To create RebHEvo4, we devised an aminoacyl tRNA synthetase based halogenase biosensor and conducted over 500 hours of phage-assisted continuous evolution (PACE). Use of RebHEvo4 in a bioreactor resulted in the production of 2.7 g/L of halogenated tryptophan. When coupled with a downstream enzyme, RebHEvo4 allowed 36-fold increased yields of halogenated tryptamines compared to the wild-type enzyme. Additionally, RebHEvo4 enabled efficient production of genetically encoded antimicrobial halogenated peptides. The efficient, site-specific halogenation by our evolved halogenase will accelerate sustainable biomanufacturing of halogenated drugs.
]]></description>
<dc:creator>Pulschen, A. A.</dc:creator>
<dc:creator>Booth, J.</dc:creator>
<dc:creator>Satanowski, A.</dc:creator>
<dc:creator>Soudy, C.</dc:creator>
<dc:creator>Caro-Astorga, J.</dc:creator>
<dc:creator>Ather, O.</dc:creator>
<dc:creator>Patel, N.</dc:creator>
<dc:creator>Alidoust, A.</dc:creator>
<dc:creator>Aoudjane, S.</dc:creator>
<dc:creator>Nematollahi, L.</dc:creator>
<dc:creator>DeBenedictis, E.</dc:creator>
<dc:date>2025-10-08</dc:date>
<dc:identifier>doi:10.1101/2025.10.08.681035</dc:identifier>
<dc:title><![CDATA[Continuous evolution of a halogenase enzyme with improved solubility and activity for sustainable bioproduction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.12.681824v1?rss=1">
<title>
<![CDATA[
Cell cycle oscillations in a polarity network facilitate state switching by morphogenetic cues 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.12.681824v1?rss=1"
</link>
<description><![CDATA[
The proper establishment of cell form, fate, and function during morphogenesis requires precise coordination between cell polarity and developmental cues. To achieve this, cells must establish polarity domains that are stable yet sensitive to guiding cues. Here we show that C. elegans germline blastomeres resolve this trade-off by creating a time-varying polarization landscape. Specifically, coupling the PAR polarity network to the cell-cycle kinase CDK-1 ensures that newborn cells operate in a low-feedback regime that lowers barriers to polarity state switching, allowing spatial cues to induce and orient PAR protein asymmetries. As CDK-1 activity rises at mitotic entry, increasing molecular feedback reinforces cue-induced asymmetries to yield robust and stable patterning of PAR domains. Consistent with this model, optogenetic and chemical perturbations show that low-CDK/low-feedback regimes destabilize PAR domains but are required for both de novo polarization and the reorientation of polarity in response to inductive cues. We propose that mitotic oscillations in cell polarity circuits dynamically optimize the polarization landscape to enable coordination of polarity with morphogenesis. Such temporal control of developmental networks is likely a general mechanism to balance robustness of cellular states with sensitivity to signal-induced state switching.



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]]></description>
<dc:creator>Ng, K.</dc:creator>
<dc:creator>Sebaa, H.</dc:creator>
<dc:creator>Hirani, N.</dc:creator>
<dc:creator>Chizh, A.</dc:creator>
<dc:creator>Messi, Z.</dc:creator>
<dc:creator>Bland, T.</dc:creator>
<dc:creator>Sugioka, K.</dc:creator>
<dc:creator>Goehring, N. W.</dc:creator>
<dc:date>2025-10-13</dc:date>
<dc:identifier>doi:10.1101/2025.10.12.681824</dc:identifier>
<dc:title><![CDATA[Cell cycle oscillations in a polarity network facilitate state switching by morphogenetic cues]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.12.681852v1?rss=1">
<title>
<![CDATA[
Extensive and differential platinum chemotherapy mutagenesis in children 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.12.681852v1?rss=1"
</link>
<description><![CDATA[
Childhood cancer survivors often develop long-term adverse effects, which may be caused by direct mutagenesis of cytotoxic agents. Some of these agents generate distinctive DNA imprints (mutational signatures), as exemplified by platinum chemotherapeutics. Here, we examined chemotherapy mutagenesis in paediatric tissues by deploying a duplex sequencing method (NanoSeq), which enables mutation calling from single DNA molecules. We surveyed whole genomes of paediatric liver, blood and other tissues, obtained from surgical resections and at post-mortem. Platinum signatures pervaded all tissues extensively, elevating mutation burdens of paediatric tissues to levels seen in adults. Remarkably, we found a tissue-specific mutational signature in the liver. We examined the functional potential of mutations by gene focused NanoSeq, which revealed that platinum agents cause a vast repertoire of cancer causing variants across normal tissues, such as leukaemogenic mutations in blood. This finding may conceivably link cancer treatment in childhood to mutation-driven long term sequelae.
]]></description>
<dc:creator>Wenger, A.</dc:creator>
<dc:creator>Lee-Six, H.</dc:creator>
<dc:creator>Dave, M.</dc:creator>
<dc:creator>Layeghifard, M.</dc:creator>
<dc:creator>Lawson, A. R. J.</dc:creator>
<dc:creator>Abascal, F.</dc:creator>
<dc:creator>Nicola, P. A.</dc:creator>
<dc:creator>Treger, T. D.</dc:creator>
<dc:creator>Ogbonnah, T.</dc:creator>
<dc:creator>Parks, C.</dc:creator>
<dc:creator>Oliver, T. R. W.</dc:creator>
<dc:creator>Kennedy, J.</dc:creator>
<dc:creator>Hodder, A.</dc:creator>
<dc:creator>Anderson, N. D.</dc:creator>
<dc:creator>Silva, F. L. T.</dc:creator>
<dc:creator>Trinh, M. K.</dc:creator>
<dc:creator>Dowe, T.</dc:creator>
<dc:creator>Habarwaa, M.</dc:creator>
<dc:creator>Sun, J. J.</dc:creator>
<dc:creator>Assia-Zamora, S.</dc:creator>
<dc:creator>Cortes-Cerisuelo, M.</dc:creator>
<dc:creator>Jassem, W.</dc:creator>
<dc:creator>Town, C.</dc:creator>
<dc:creator>Dhawan, A.</dc:creator>
<dc:creator>Jain, V.</dc:creator>
<dc:creator>Straathof, K.</dc:creator>
<dc:creator>Deheragoda, M.</dc:creator>
<dc:creator>Martincorena, I.</dc:creator>
<dc:creator>Palm, L.</dc:creator>
<dc:creator>Hutchinson, J. C.</dc:creator>
<dc:creator>Coorens, T. H. H.</dc:creator>
<dc:creator>Trayers, C.</dc:creator>
<dc:creator>Heaton, N.</dc:creator>
<dc:creator>Shlien, A.</dc:creator>
<dc:creator>Zen, Y.</dc:creator>
<dc:creator>Rouhani, F. J.</dc:creator>
<dc:creator>Behjati, S.</dc:creator>
<dc:date>2025-10-14</dc:date>
<dc:identifier>doi:10.1101/2025.10.12.681852</dc:identifier>
<dc:title><![CDATA[Extensive and differential platinum chemotherapy mutagenesis in children]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.13.681913v1?rss=1">
<title>
<![CDATA[
Ecological cues orchestrate concerted courtship in a Drosophila host specialist 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.13.681913v1?rss=1"
</link>
<description><![CDATA[
Mating decisions are often attributed to the sensory signaling between prospective sexual partners. Yet these interactions are also shaped by the broader environmental context in which they unfold, to appropriately align sexual arousal with reproductive opportunities. Here we show that in the host specialist Drosophila erecta mating is strictly contingent on the ecological and social environment generated as flies densely aggregate in groups on a food patch. We find that food volatiles directly promote male sexual arousal, triggering individuals to sample and pursue potential mates, giving rise to dynamic interactions across the group. The ensuing visual motion transforms each males visual field, which in turn further amplifies his arousal, generating a multisensory feedback loop that coordinately promotes courtship across individuals. D. erectas strict dependence on environmental cues appears latent in related species, such as D. melanogaster, where food odor can promote arousal but is dispensable for vigorous courtship. Comparative circuit analyses reveal that species-specific thresholds for sexual arousal reflect variation in how olfactory input modulates conserved nodes controlling courtship drive, rendering food volatiles a strict sensory gate only in D. erecta. Together, our findings highlight how ecological cues not directly tied to sexual signaling can profoundly influence reproductive behavior and reorganize the social landscape to ensure mating occurs in contexts where reproductive opportunities are abundant.
]]></description>
<dc:creator>Brand, P.</dc:creator>
<dc:creator>Keller, K.</dc:creator>
<dc:creator>Coleman, R. T.</dc:creator>
<dc:creator>Eghbali, N. B.</dc:creator>
<dc:creator>Zylka, S.</dc:creator>
<dc:creator>Prieto-Godino, L. L.</dc:creator>
<dc:creator>Ruta, V.</dc:creator>
<dc:date>2025-10-14</dc:date>
<dc:identifier>doi:10.1101/2025.10.13.681913</dc:identifier>
<dc:title><![CDATA[Ecological cues orchestrate concerted courtship in a Drosophila host specialist]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.14.682049v1?rss=1">
<title>
<![CDATA[
The origin and molecular evolution of the mammalian liver cell architecture 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.14.682049v1?rss=1"
</link>
<description><![CDATA[
The liver is a central organ with essential roles in processes such as nutritional metabolism, detoxification, and immune defense1-6. It has been instrumental in the adaptation of mammalian species to diverse environments, as reflected by its rapid molecular evolution7,8. However, the origins and evolutionary dynamics of liver cell types and their structural organization remain largely unexplored. Here we report evolutionary analyses of transcriptome and chromatin accessibility data for liver cells from 17 species, spanning all major feeding strategies (herbivory, omnivory, carnivory, insectivory), great apes (including humans), placental clades (Afrotheria, Xenarthra, Laurasiatheria, Euarchontoglires), major mammalian lineages (placentals, marsupials, monotremes), and a bird as outgroup. Integrated with spatial transcriptomics, our data reveal that liver zonation--the compartmentalization of hepatocyte functions along the lobule, the livers fundamental anatomical and functional unit--is conserved across mammals but absent in other vertebrates. We find that zonation originated in the mammalian ancestor, driven by the emergence of WNT and R-spondin signaling from central vein endothelial cells, which activate central hepatocyte gene expression via the transcription factor TCF7L2. Despite this conserved architecture and signaling, genes with zonated expression exhibit rapid evolutionary turnover. Consistently, hepatocytes evolve fast, likely due to reduced selective constraints, enabling adaptive changes under positive selection. Alongside immune cells, hepatocytes are therefore key drivers of the livers rapid evolution and functional innovations. In great apes, we identify human-specific shifts in zonation and cell-type-specific expression linked to recent cis-regulatory changes, particularly in genes involved in lipid metabolism, likely contributing to human-specific metabolic traits. Our study uncovers the origins of a mammal-specific liver cell architecture, within which reduced constraints facilitated molecular changes underlying ecological adaptations.
]]></description>
<dc:creator>Yuan, X.</dc:creator>
<dc:creator>Rodriguez-Montes, L.</dc:creator>
<dc:creator>Zaremba, B.</dc:creator>
<dc:creator>Trost, N.</dc:creator>
<dc:creator>Schneider, C.</dc:creator>
<dc:creator>Schmidt, J.</dc:creator>
<dc:creator>Berki, B.</dc:creator>
<dc:creator>Ibberson, D.</dc:creator>
<dc:creator>Leushkin, E.</dc:creator>
<dc:creator>Nickel, B.</dc:creator>
<dc:creator>Palkovits, M.</dc:creator>
<dc:creator>Truman, R. W.</dc:creator>
<dc:creator>Barsh, G.</dc:creator>
<dc:creator>Lees, J.</dc:creator>
<dc:creator>Fallahshahroudi, A.</dc:creator>
<dc:creator>Grützner, F.</dc:creator>
<dc:creator>Tzika, A. C.</dc:creator>
<dc:creator>Milinkovitch, M. C.</dc:creator>
<dc:creator>Pääbo, S.</dc:creator>
<dc:creator>Cardoso-Moreira, M.</dc:creator>
<dc:creator>Kaessmann, H.</dc:creator>
<dc:date>2025-10-14</dc:date>
<dc:identifier>doi:10.1101/2025.10.14.682049</dc:identifier>
<dc:title><![CDATA[The origin and molecular evolution of the mammalian liver cell architecture]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.13.682089v1?rss=1">
<title>
<![CDATA[
The SNCA A53T mutation sensitizes human neurons and microglia to ferroptosis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.13.682089v1?rss=1"
</link>
<description><![CDATA[
The major pathological hallmarks of sporadic and familial forms of Parkinsons disease (PD) are the targeted and progressive loss of midbrain dopaminergic neurons (mDA), associated with systemic iron accumulation, -synuclein (syn) accumulation and aggregation, and lipid peroxidation amongst other reactive oxygen species (ROS) generation. Therapeutic strategies aimed towards dopamine restoration, syn removal and iron chelation have provided symptomatic relief but failed to prevent or slow disease progression. This is in part due to the lack of understanding of the exact pathways leading to neuronal death in PD. In this study, we investigate ferroptosis, a unique cell death mechanism sharing multiple features with PD pathology, as a relevant pathway with implications in disease pathogenesis. We identified an enrichment of ferroptosis genes dysregulated throughout PD postmortem brain samples and several neuronal and glial PD models. Using CRISPR/Cas9 technology, we generated a rapid iPSC-derived synucleinopathy neuronal model harbouring the SNCA A53T mutation and report increased ROS generation, reduced levels of antioxidant glutathione (GSH), impaired mitophagy and a heightened vulnerability to ferroptosis-induced lipid peroxidation and cell death. Critically, inhibition of the key lipid peroxidation enzyme and driver of ferroptosis, 15-lipoxygenase (15-LO), rescued synucleinopathy associated pathologies and prevented pathological syn oligomerisation in SNCA A53T neurons. Furthermore, we report enhanced microglial ferroptosis susceptibility in models of synucleinopathy. In summary, we highlight a new mechanism by which the familial PD-associated SNCA A53T mutation causes cell death and propose 15-LO inhibition as a tractable therapeutic opportunity in PD.
]]></description>
<dc:creator>Mahoney-Sanchez, L.</dc:creator>
<dc:creator>Clarke-Lucas, H.</dc:creator>
<dc:creator>Penverne, A.</dc:creator>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>D'Sa, K.</dc:creator>
<dc:creator>Strohbuecker, S.</dc:creator>
<dc:creator>Lopex Garcia, P.</dc:creator>
<dc:creator>Cosker, K.</dc:creator>
<dc:creator>Soltic, D.</dc:creator>
<dc:creator>O'Callaghan, B. J.</dc:creator>
<dc:creator>Griffiths, A.</dc:creator>
<dc:creator>Pintchovski, S. A.</dc:creator>
<dc:creator>Plun-Favreau, H.</dc:creator>
<dc:creator>Hallqvist, J.</dc:creator>
<dc:creator>Mills, K.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:date>2025-10-14</dc:date>
<dc:identifier>doi:10.1101/2025.10.13.682089</dc:identifier>
<dc:title><![CDATA[The SNCA A53T mutation sensitizes human neurons and microglia to ferroptosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.20.683393v1?rss=1">
<title>
<![CDATA[
Autoantibody origins in lupus and in relapse post CAR-T therapy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.20.683393v1?rss=1"
</link>
<description><![CDATA[
Anti-CD19 chimeric antigen receptor (CAR)-T therapy induces profound remissions in lupus by depleting B cells, challenging the longstanding view that treatment-resistant disease is sustained by long-lived plasma cells. Additionally, emerging relapses highlight the need to understand how pathogenic autoantibodies arise. Using molecular antibody tagging in mice with human monogenic lupus variants, we reveal that autoantibody-producing cell cohorts are long-lived but plasma cells are short-lived, requiring continuous replenishment from proliferating precursors. The spleen acts as a major plasma cell reservoir, with perivascular localization conserved in mice and lupus patients. Relapse after anti-CD19 CAR-T occurred through newly-generated B cells rather than treatment-resistant clones. Plasma cell depletion by anti-BCMA CAR-T failed to eliminate some precursors that become autoantibody-secreting. These findings demonstrate that continuous B cell-to-plasma cell differentiation, not intrinsic plasma cell longevity, sustains pathogenic antibody responses in lupus, supporting a potential benefit of adjunctive therapies after CAR-T, particularly in genetically predisposed patients.
]]></description>
<dc:creator>Grenov, A.</dc:creator>
<dc:creator>Yoon, J.</dc:creator>
<dc:creator>Snell, D. M.</dc:creator>
<dc:creator>Mikolajczak, A.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Gurel, E.</dc:creator>
<dc:creator>Rodriguez-Ronchel, A.</dc:creator>
<dc:creator>Yegen, G.</dc:creator>
<dc:creator>Lee, L.</dc:creator>
<dc:creator>Artim Esen, B.</dc:creator>
<dc:creator>Rahman, A.</dc:creator>
<dc:creator>Maciocia, P.</dc:creator>
<dc:creator>Vinuesa, C.</dc:creator>
<dc:date>2025-10-20</dc:date>
<dc:identifier>doi:10.1101/2025.10.20.683393</dc:identifier>
<dc:title><![CDATA[Autoantibody origins in lupus and in relapse post CAR-T therapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.22.684057v1?rss=1">
<title>
<![CDATA[
Inhibition of neutrophil degranulation by Nexinhib20 delays the development of radiation-induced pulmonary fibrosis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.22.684057v1?rss=1"
</link>
<description><![CDATA[
Despite advances in radiation delivery techniques that enhance tumour targeting and minimise collateral exposure, healthy tissues remain susceptible to radiation-induced fibrosis, a chronic and progressive condition that can severely compromise patient quality of life. Therapeutic options to prevent or treat radiation-induced fibrosis remain limited. Recent work has shown that neutrophils infiltrating healthy lung tissue after irradiation adopt an activated phenotype capable of perturbing cellular responses of both epithelial and mesenchymal cells. However, the contribution of these radiation-educated neutrophils to the development of radiation-induced fibrosis remains unclear. Using targeted, image-guided lung irradiation to deliver a dose sufficient to induce fibrosis within four months, we demonstrate that neutrophils are essential for the efficient development of clinically evident fibrosis. Lung irradiation educates neutrophils, enabling them to promote early alterations in the extracellular matrix shortly after radiation exposure. We further show that this "educated" phenotype depends on neutrophil degranulation activity. Pharmacological inhibition of degranulation with Nexinhib20 redirected these cells toward a pro-angiogenic and anti-fibrotic phenotype. Importantly, this treatment was also associated with a transcriptional shift in mesenchymal cells away from a pro-fibrotic program, resulting in a marked delay in the onset of radiation-induced fibrosis. Importantly, Nexinhib20 did not impair the efficacy of cancer radiotherapy, underscoring its potential as a therapeutic strategy to prevent fibrotic complications without diminishing anti-tumour effectiveness.
]]></description>
<dc:creator>Ruhland, S.</dc:creator>
<dc:creator>Nikolatou, K.</dc:creator>
<dc:creator>Bridgeman, V. L.</dc:creator>
<dc:creator>Rizou, T.</dc:creator>
<dc:creator>Sahai, E.</dc:creator>
<dc:creator>Bordoloi, J.</dc:creator>
<dc:creator>Nolan, E.</dc:creator>
<dc:creator>Elosegui-Artola, A.</dc:creator>
<dc:creator>Malanchi, I.</dc:creator>
<dc:date>2025-10-23</dc:date>
<dc:identifier>doi:10.1101/2025.10.22.684057</dc:identifier>
<dc:title><![CDATA[Inhibition of neutrophil degranulation by Nexinhib20 delays the development of radiation-induced pulmonary fibrosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.20.683483v1?rss=1">
<title>
<![CDATA[
Cancer-driven neutrophil priming couples systemic epithelial regenerative programs with pre-metastatic niche formation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.20.683483v1?rss=1"
</link>
<description><![CDATA[
Cancer progression involves systemic changes that extend beyond the primary tumour. Through cancer-induced systemic conditioning, breast tumours generate subclinical alterations in distant organs that facilitate metastatic seeding and pre-metastatic niche formation. Neutrophils, mobilized through cancer-driven emergency granulopoiesis, actively contribute to this process. In this study we extend the concept of neutrophil-dependent conditioning beyond pre-metastatic sites, uncovering a broader systemic regenerative activation that links inflammation, tissue regeneration, and metastasis. This activation manifests as enhanced epithelial progenitor activity, measured by increased organoid formation, across multiple organs, including those with low risk of breast cancer metastasis. This neutrophil-dependent perturbation in lung alveolar progenitors and intestinal epithelial lineage commitment, is an indication of an altered organ physiology, enhancing tissue resilience to injury. Moreover, we identify UPP1 expression, which exclusively characterizes neutrophils generated through emergency granulopoiesis, as a key factor sustaining high translational activity in neutrophil progenitors and enabling the full acquisition of cancer-primed properties. Consequently, neutrophil loss of UPP1 reduces both their lung pro-metastatic function and their capacity to activate alveolar progenitors. Mechanistically, this involves interactions between cancer-primed neutrophils and platelets, which localize within lung interstitial spaces near alveolar cells to stimulate epithelial progenitor activity.

Together, these findings uncover a previously unrecognized tumour-induced systemic conditioning in which neutrophils coordinate epithelial regenerative activation as part of a pro-metastatic epithelial niche, with UPP1 as a key determinant of their cancer-primed state.
]]></description>
<dc:creator>Rabas, N.</dc:creator>
<dc:creator>Bridgeman, V.</dc:creator>
<dc:creator>Hennequart, M.</dc:creator>
<dc:creator>Rodrigues, F. S.</dc:creator>
<dc:creator>Farias, A.</dc:creator>
<dc:creator>Ferreira, R. M. M.</dc:creator>
<dc:creator>Axarlis, K.</dc:creator>
<dc:creator>Nduka, O.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Legrave, N.</dc:creator>
<dc:creator>Furlani, I.</dc:creator>
<dc:creator>Malanchi, I.</dc:creator>
<dc:date>2025-10-21</dc:date>
<dc:identifier>doi:10.1101/2025.10.20.683483</dc:identifier>
<dc:title><![CDATA[Cancer-driven neutrophil priming couples systemic epithelial regenerative programs with pre-metastatic niche formation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.30.685573v1?rss=1">
<title>
<![CDATA[
Post-infection pig and ferret antisera show similar antigenic profiles for human influenza A(H1N1pdm09) viruses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.30.685573v1?rss=1"
</link>
<description><![CDATA[
BackgroundMonitoring antigenic drift in human influenza A viruses is essential for vaccine strain selection and ensuring protection against circulating strains. Antigenic drift is traditionally assessed using ferret antisera, which provide monospecific responses, and human vaccinee sera, which reflect exposure to multiple antigens. In this study we evaluated the pig as an alternative source of antisera to study antigenic drift compared to immune responses in ferrets and humans. We included seasonal influenza A(H1N1pdm09) human viruses that had shown different antigenic characteristics when using ferret or human antisera.

MethodsPairs of pigs were inoculated with six human A(H1N1)pdm09 viruses circulating between 2019 and 2023, a period of marked antigenic drift. Pig and ferret antisera were analysed by hemagglutination inhibition (HI) and virus neutralization (VN) assays.

ResultsPigs were successfully infected with all strains, shedding virus and producing antibody responses, confirming their susceptibility to human influenza A viruses. Antigenic reactivity of pig antisera was qualitatively comparable to ferret antisera in both HI and VN assays, although maximum homologous antibody titres were significantly higher in ferrets. The antisera raised against viruses in circulation in 2019 and before, exempified by A/Guangdong-Maonan/SWL1536/2019, clade 5a.1, were clearly differentiated by both ferret and pig antisera from those in clade 5a.2 and its derivatives that became predominant.

ConclusionsFerrets and pigs showed comparable responses and both distinguished clade 5a.1 from clade 5a.2. However, neither model recognised antigenically drifted variants from 2019-2022, including subclades 5a.2-C, 5a.2a-C.1/C.1.9, and .5a.2a.1-C.1.1/D, which were distinguishable using human post-vaccination antisera.
]]></description>
<dc:creator>Harvey, R.</dc:creator>
<dc:creator>Cayol, T.</dc:creator>
<dc:creator>Paudyal, B.</dc:creator>
<dc:creator>Lilley, A.</dc:creator>
<dc:creator>Carr, C.</dc:creator>
<dc:creator>Hatton, C.</dc:creator>
<dc:creator>Briggs, E.</dc:creator>
<dc:creator>Daniels, R. S.</dc:creator>
<dc:creator>Richardson, S.</dc:creator>
<dc:creator>Peacock, T. P.</dc:creator>
<dc:creator>Lewis, N. S.</dc:creator>
<dc:creator>Brown, I.</dc:creator>
<dc:creator>Mccauley, J.</dc:creator>
<dc:creator>Tchilian, E.</dc:creator>
<dc:date>2025-10-31</dc:date>
<dc:identifier>doi:10.1101/2025.10.30.685573</dc:identifier>
<dc:title><![CDATA[Post-infection pig and ferret antisera show similar antigenic profiles for human influenza A(H1N1pdm09) viruses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.28.685054v1?rss=1">
<title>
<![CDATA[
Regulation of cell proliferation by a novel feedback system on Cdk function 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.28.685054v1?rss=1"
</link>
<description><![CDATA[
The proliferation of eukaryotic cells is regulated by a complex network of regulatory systems that promotes efficient cell cycle progression and ensures proper responses to the environment. Despite this complexity, the core inputs that are necessary and sufficient for robust alternation of DNA replication and mitosis are surprisingly simpler than anticipated. Indeed, fission yeast cells operating with an engineered minimal cell cycle network that lacks the highly conserved Wee1+Cdc25 feedback loops on Cdk1 function are viable, although slow growing. This provides a unique entry for evaluating how such simplified cells can evolve and improve their proliferation potential while exploring unknown mechanisms modulating cell cycle progression. Taking advantage of this model, we applied laboratory evolution assays to minimal fission yeast backgrounds and selected for the emergence of faster growing populations. We found that loss of the small disordered protein Spo12 brings about enhanced population growth in cells lacking the Wee1+Cdc25 mitotic switch. Importantly, we demonstrate that Spo12 defines a new and conserved family of inhibitors of the Cdk-counteracting phosphatase PP2A that are directly regulated by Cdk-dependent phosphorylation. Our results also reveal a trade-off associated with Spo12-dependent regulation, which may have implications for our understanding of the principles underlying the evolution of cell cycle control. Finally, our study highlights how combining simplified circuits with experimental evolution allows for uncovering regulatory elements that may be obscured by network complexity.
]]></description>
<dc:creator>Ryan, J. R.</dc:creator>
<dc:creator>Leray, B.</dc:creator>
<dc:creator>Jain, A.</dc:creator>
<dc:creator>Coehlo, N.</dc:creator>
<dc:creator>Domingo-Sananes, M. R.</dc:creator>
<dc:creator>Aspert, T.</dc:creator>
<dc:creator>Charvin, G.</dc:creator>
<dc:creator>Krockenberger, M.</dc:creator>
<dc:creator>Chica Blaguera, N.</dc:creator>
<dc:creator>Lopez Aviles, S.</dc:creator>
<dc:creator>Takacs, L.</dc:creator>
<dc:creator>Uhlmann, F.</dc:creator>
<dc:creator>Yang, L.</dc:creator>
<dc:creator>Yue, J.-X.</dc:creator>
<dc:creator>Liti, G.</dc:creator>
<dc:creator>Cochard, V.</dc:creator>
<dc:creator>Chevreux, G.</dc:creator>
<dc:creator>Sugiyama, H.</dc:creator>
<dc:creator>Goto, Y.</dc:creator>
<dc:creator>Aoki, K.</dc:creator>
<dc:creator>Wu, P.-Y. J.</dc:creator>
<dc:creator>Mackereth, C. D.</dc:creator>
<dc:creator>Tyson, J.</dc:creator>
<dc:creator>Novak, B.</dc:creator>
<dc:creator>Coudreuse, D.</dc:creator>
<dc:date>2025-10-29</dc:date>
<dc:identifier>doi:10.1101/2025.10.28.685054</dc:identifier>
<dc:title><![CDATA[Regulation of cell proliferation by a novel feedback system on Cdk function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.28.685133v1?rss=1">
<title>
<![CDATA[
Transcript-Capture sequencing enriches mRNA of Mycobacterium tuberculosis from host samples 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.28.685133v1?rss=1"
</link>
<description><![CDATA[
Bacterial gene expression from sites of infection are poorly studied due to low levels of bacterial mRNA present in clinical samples. Here, we develop Transcript-Capture Seq, which uses customizable biotinylated probes generated in-house to enrich bacteria-specific RNA from host samples before Next Generation Sequencing (NGS). This method results in a >200-fold increase in bacterial mRNA reads from mixed samples and allows analysis of the complete bacterial transcriptome from clinical samples. We apply Transcript-Capture to models of tuberculosis (TB) infection as well as sputum samples from TB patients. TB patients exhibit unexplained heterogeneity in disease progression, and the activity of Mycobacterium tuberculosis (Mtb) has been proposed to affect treatment response. By applying Transcript-Capture to sputum samples collected from TB patients we generate the first complete in vivo bacterial transcriptome of Mtb via NGS. Mtb from patient sputa shows upregulation of genes involved in host lipid utilization and zinc limitation, as well as a similar gene expression profile to Mtb log phase growth in vitro. Applying Transcript-Capture to clinical sputa provides a snapshot of bacterial activity directly from human patients and can be used to investigate the physiological state of bacteria surviving in vivo.

GRAPHICAL ABSTRACT

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=74 SRC="FIGDIR/small/685133v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Lamont, E. I.</dc:creator>
<dc:creator>Jones, R. M.</dc:creator>
<dc:creator>Assadi, J.</dc:creator>
<dc:creator>Morrison, R.</dc:creator>
<dc:creator>Song, T.</dc:creator>
<dc:creator>Yu, X.</dc:creator>
<dc:creator>Wiener, D. M.</dc:creator>
<dc:creator>Via, L. E.</dc:creator>
<dc:creator>Winter, J.</dc:creator>
<dc:creator>Ma, S. I.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Barry, C. E.</dc:creator>
<dc:creator>Sherman, D. R.</dc:creator>
<dc:date>2025-10-28</dc:date>
<dc:identifier>doi:10.1101/2025.10.28.685133</dc:identifier>
<dc:title><![CDATA[Transcript-Capture sequencing enriches mRNA of Mycobacterium tuberculosis from host samples]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.27.684825v1?rss=1">
<title>
<![CDATA[
Leveraging death of drug-sensitive cancer cells to promote immune-mediated bystander killing of subclones of drug-resistant tumor cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.27.684825v1?rss=1"
</link>
<description><![CDATA[
The response of lung cancer patients to drugs targeting the G12C mutant form of KRAS is limited by the development of resistance through multiple mechanisms. In order to achieve lasting benefit with these therapies, effective strategies for tackling the evolution of drug resistance are required. We have developed a preclinical model system to mimic the development of resistance to KRAS G12Ci inhibitors (G12Ci) such as adagrasib and RMC-4998. Treatment of tumours containing a minor subpopulation of resistant cancer cells with G12Ci leads to their rapid outgrowth to replace the drug-sensitive cells within a few weeks. However, when combined with therapies that, at least in part, target the immune response, such as SHP2 inhibitors or PD-1 blockade, drug-resistant cells can be eliminated, even by drug combinations that do not impact their growth in the absence of drug-sensitive cells. This bystander killing of drug-resistant cells when drug-sensitive cells are targeted is dependent on an intact adaptive immune system. Mechanistically, these combination therapies lead to profound remodelling of the tumour immune microenvironment, with influx of T cells recognising a tumour associated antigen shared between drug-resistant and drug-sensitive cancer cells. Promotion of immune-mediated bystander killing of drug-resistant cells may provide a paradigm for tackling the problem of drug resistance in cancer more broadly.
]]></description>
<dc:creator>Tomaschko, M.</dc:creator>
<dc:creator>Ng, K.</dc:creator>
<dc:creator>Moore, C.</dc:creator>
<dc:creator>Pillsbury, C. E.</dc:creator>
<dc:creator>Rana, S.</dc:creator>
<dc:creator>Campbell, J.</dc:creator>
<dc:creator>Mukherjee, S.</dc:creator>
<dc:creator>Mikolajczak, A.</dc:creator>
<dc:creator>Anastasiou, P.</dc:creator>
<dc:creator>de Castro, A.</dc:creator>
<dc:creator>Alonso de la Vega, A.</dc:creator>
<dc:creator>de Carne Trecesson, S.</dc:creator>
<dc:creator>Goehring, N.</dc:creator>
<dc:creator>Molina-Arcas, M.</dc:creator>
<dc:creator>Downward, J.</dc:creator>
<dc:date>2025-10-28</dc:date>
<dc:identifier>doi:10.1101/2025.10.27.684825</dc:identifier>
<dc:title><![CDATA[Leveraging death of drug-sensitive cancer cells to promote immune-mediated bystander killing of subclones of drug-resistant tumor cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.27.684794v1?rss=1">
<title>
<![CDATA[
SERINC5 Is the Primary Target of Retroviral Antagonists Among Human SERINC Paralogs and Requires a Tri-Cysteine Motif for Its Counteraction by HIV-1 Nef 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.27.684794v1?rss=1"
</link>
<description><![CDATA[
The human transmembrane protein SERINC5 incorporates into budding HIV-1 particles and inhibits their fusion with new target cells. Among the five members of the SERINC family, SERINC3 and SERINC1 exhibit modest antiviral activity, whereas SERINC2 lacks this function. HIV-1 counteracts SERINC5 by promoting its endocytosis in a Nef- dependent manner. SERINC5 is also downregulated by glycoGag of Moloney murine leukaemia virus (MoMLV) and by the S2 glycoprotein of equine infectious anaemia virus (EIAV).

Here, we demonstrate that all human SERINCs, despite their differing antiviral potencies, are incorporated into HIV-1 virions. SERINC5 is the most sensitive to viral antagonists, whereas its paralogs display moderate (SERINC2, SERINC3) or no (SERINC1) susceptibility, suggesting that SERINC5 has exerted stronger selective pressure on retroviral evolution.

To identify the determinants of SERINC5 sensitivity to Nef, we performed structure- guided mutagenesis and identified a critical tri-cysteine motif (Cys355, Cys356, Cys358) within intracellular loop 4 (ICL4) required for Nef-mediated downregulation. Substitution of these residues with glutamines conferred resistance to Nef antagonism. While this motif is essential for SERINC5 internalization by diverse HIV-1 and SIV Nef alleles, it is dispensable for downregulation by MoMLV glycoGag and EIAV S2, indicating that these viral antagonists exploit distinct structural features of SERINC5. Finally, we show that SERINC5 is palmitoylated at the tri-cysteine motif, raising the possibility that this modification modulates its sensitivity to Nef.

Together, these results confirm the central role of SERINC5 as a target of retroviral counteraction and highlight the crucial importance of ICL4 in mediating its engagement by Nef.

ImportanceThe SERINC family of multipass transmembrane proteins includes host factors capable of restricting retrovirus infectivity. Among the five human paralogs, SERINC5 displays the strongest antiviral activity and is the only member efficiently targeted by all known retroviral antagonists, including HIV-1 Nef, MLV glycoGag, and the EIAV S2. This selectivity indicates that the evolutionary pressure exerted by SERINC proteins on retroviruses has been primarily driven by SERINC5 rather than by its relatives. Through structure-guided mutagenesis, we identified a tri-cysteine motif within intracellular loop 4 as a crucial determinant for SERINC5 downregulation by Nef, providing a molecular explanation for its distinctive susceptibility. These findings establish SERINC5 as the principal target of viral counteraction and suggest that its molecular features have contributed to shaping the adaptation of primate lentiviral Nef proteins to the host.
]]></description>
<dc:creator>Bertelli, C.</dc:creator>
<dc:creator>Rosa, A.</dc:creator>
<dc:creator>Vanzo, T.</dc:creator>
<dc:creator>Chande, A.</dc:creator>
<dc:creator>Serwa, R.</dc:creator>
<dc:creator>Tate, E. W.</dc:creator>
<dc:creator>Cherepanov, P. P.</dc:creator>
<dc:creator>Pizzato, M.</dc:creator>
<dc:date>2025-10-27</dc:date>
<dc:identifier>doi:10.1101/2025.10.27.684794</dc:identifier>
<dc:title><![CDATA[SERINC5 Is the Primary Target of Retroviral Antagonists Among Human SERINC Paralogs and Requires a Tri-Cysteine Motif for Its Counteraction by HIV-1 Nef]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.23.684281v1?rss=1">
<title>
<![CDATA[
Multicellular Calcium Waves in Cancer-Associated Fibroblasts Regulate Neuronal Mimicry and Anisotropy Leading to Immune Exclusion 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.23.684281v1?rss=1"
</link>
<description><![CDATA[
Stromal barriers exclude CD8+ T cells from accessing cancer cells and hamper immune-mediated tumour control. Through multi-pronged analysis of tumours that transition from immune inflamed to immune excluded, we reveal that the formation of stromal barriers is associated with the acquisition of neuronal gene expression programmes in cancer-associated fibroblasts (CAFs), including TUBB3 expression. This leads to neuronal mimicry, with stromal barrier formation underpinned by coordinated transient bursts of intracellular calcium release, similar to those observed in neuronal tissue. Blockade of calcium release through either pharmacological or molecular interventions, such as STC2 depletion, prevents CAF alignment and the build-up of CD8+ T cells at stromal boundaries. Nintedanib treatment prevents neuronal mimicry and restores immune-mediated tumour control. Thus, we uncover unexpected mimicry of neuronal behaviour in CAFs, document the mechanism by which it leads to immune exclusion, and identify ways to prevent the induction of neuronal mimicry and restore immune-mediated tumour control.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=95 SRC="FIGDIR/small/684281v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Giangreco, G.</dc:creator>
<dc:creator>Ramsden, Z.</dc:creator>
<dc:creator>Rullan, A.</dc:creator>
<dc:creator>Hooper, S.</dc:creator>
<dc:creator>Novo, D.</dc:creator>
<dc:creator>Castro, J. L.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Milan, M.</dc:creator>
<dc:creator>Mohammadi, H.</dc:creator>
<dc:creator>Llop, S.</dc:creator>
<dc:creator>Juarez, M.</dc:creator>
<dc:creator>Schneider-Luftman, D.</dc:creator>
<dc:creator>Oliva, M.</dc:creator>
<dc:creator>Alemany, L.</dc:creator>
<dc:creator>Angelova, M.</dc:creator>
<dc:creator>Olow, A.</dc:creator>
<dc:creator>Yu, X.</dc:creator>
<dc:creator>Hobson, P.</dc:creator>
<dc:creator>Harrington, K. J.</dc:creator>
<dc:creator>Sahai, E.</dc:creator>
<dc:date>2025-10-24</dc:date>
<dc:identifier>doi:10.1101/2025.10.23.684281</dc:identifier>
<dc:title><![CDATA[Multicellular Calcium Waves in Cancer-Associated Fibroblasts Regulate Neuronal Mimicry and Anisotropy Leading to Immune Exclusion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.23.684170v1?rss=1">
<title>
<![CDATA[
Cooperative and antagonistic interactions between sub-clones favour the co-existence of multiple resistance mechanisms in melanoma 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.23.684170v1?rss=1"
</link>
<description><![CDATA[
Resistance to targeted therapies develops in the majority of patients, with numerous different mechanisms having been documented. This is frequently linked to intra-tumour heterogeneity and cancer cells transitioning into different states. In this study, we demonstrate cooperativity between co-occurring resistant states in a single tumour. Using BRAF mutant melanoma as a paradigm, we generate three different resistant states within a single model and demonstrate that they have varying differentiation states, capacities to migrate, and have very few common additional therapeutic vulnerabilities. Through a combination of experiments, including using Cre-mediated recombination to generate heterogeneity in existing tumours, and in silico modelling, we show that intra-tumour heterogeneity is the most favoured state for therapy resistant tumours. This is under-pinned by signalling between different melanoma states, with YAP1 active cells providing supporting signals for other cells but inhibiting their own proliferation. Optimal disease control requires targeting both the YAP1 active cell state and the inter-cellular communication networks. We identify the histone demethylase inhibitor GSK-J4 as being particularly effective in targeting both features of resistant tumours, and demonstrate its ability to control melanoma with multiple concurrent resistance mechanisms.
]]></description>
<dc:creator>Schlegelmilch, K.</dc:creator>
<dc:creator>Hollek, V.</dc:creator>
<dc:creator>Hooper, S.</dc:creator>
<dc:creator>Giangreco, G.</dc:creator>
<dc:creator>Bailey, S.</dc:creator>
<dc:creator>Macfarlane, S.</dc:creator>
<dc:creator>Carminati, A.</dc:creator>
<dc:creator>Bowes, A.</dc:creator>
<dc:creator>Strohbuecker, S.</dc:creator>
<dc:creator>Fu, X.</dc:creator>
<dc:creator>Sahai, E.</dc:creator>
<dc:date>2025-10-24</dc:date>
<dc:identifier>doi:10.1101/2025.10.23.684170</dc:identifier>
<dc:title><![CDATA[Cooperative and antagonistic interactions between sub-clones favour the co-existence of multiple resistance mechanisms in melanoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.24.684328v1?rss=1">
<title>
<![CDATA[
Hierarchical lineage architecture of human and avian spinal cord revealed by single-cell genomic barcoding 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.24.684328v1?rss=1"
</link>
<description><![CDATA[
The formation of neural circuits depends on the precise spatial and temporal organisation of neuronal populations during development. In the vertebrate spinal cord, progenitors are patterned into molecularly defined domains, but how lineage relationships shape neuronal diversity and function has remained unclear. Here, we combine genomic barcoding with single- cell RNA sequencing in chick and human embryos to generate cell-type-resolved clonal maps. We find that spinal neurogenesis follows a hierarchical organisation in which the neural tube first partitions into five broad subdivisions that resolve into the eleven progenitor domains generating the cardinal neuronal classes. This bifurcating architecture implies a patterning mechanism of sequential binary decisions. The most prominent lineage restriction occurs at the embryonic alar-basal boundary, separating sensory-processing from motor-control circuits. Individual progenitors generate neurons across multiple temporal waves while remaining constrained within their lineage subdivision, demonstrating persistence of spatial identity despite temporal competence changes. Among sensory populations, we identify two developmental routes, via unifated or bifated progenitors, to pain- and itch-processing interneurons. These principles are conserved between chick and human, with clonal analysis in human embryos revealing that most fate choices are resolved by six weeks post-conception. Together, these findings provide a framework for spinal cord development and reveal lineage compartmentalisation as a fundamental principle in neural circuit assembly and evolution.
]]></description>
<dc:creator>Boezio, G. L. M.</dc:creator>
<dc:creator>Depotter, J. R. L.</dc:creator>
<dc:creator>Frith, T. J. R.</dc:creator>
<dc:creator>Radley, A.</dc:creator>
<dc:creator>Strohbuecker, S.</dc:creator>
<dc:creator>Cunha, A. C.</dc:creator>
<dc:creator>Howell, M.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:date>2025-10-24</dc:date>
<dc:identifier>doi:10.1101/2025.10.24.684328</dc:identifier>
<dc:title><![CDATA[Hierarchical lineage architecture of human and avian spinal cord revealed by single-cell genomic barcoding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.09.686930v1?rss=1">
<title>
<![CDATA[
Spillover of H5 influenza viruses to vampire bats at the marine-terrestrial interface 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.09.686930v1?rss=1"
</link>
<description><![CDATA[
The highly pathogenic H5N1 avian influenza A virus (IAV) clade 2.3.4.4b has spread globally and spilled over into multiple mammalian species, raising concerns about its pandemic potential. In late 2022, clade 2.3.4.4b viruses devastated seabird and marine mammal populations along the Pacific coast of South America. Here, we report the first evidence of H5 IAV infections in wild bats globally, focusing on common vampire bats (Desmodus rotundus) in coastal areas of Peru. Longitudinal serological screening, stable isotope analysis and metabarcoding revealed repeated exposures to H5 IAVs in vampire bats which feed on coastal wildlife species heavily impacted by the 2.3.4.4b epizootic, but no evidence of infection in populations without access to marine prey. We further report bat gene flow between IAV-exposed and IAV-naive populations, and IAV infections in a vampire bat colony that fed on both marine and terrestrial livestock prey, providing insights into how future IAV epizootics might spread spatially within bats and between marine and terrestrial ecosystems if a bat reservoir were established. Immunohistochemistry demonstrated that the H5 haemagglutinin protein binds to the upper respiratory tract of vampire bats, suggesting bat tissue susceptibility to H5 IAVs. Finally, vampire bat-derived kidney, liver, and lung cells supported entry, replication, and egress of avian and mammalian 2.3.4.4b viruses, confirming cellular infectivity. These results illustrate how combining ecological inference and experimental virology can pinpoint the species origins and biological significance of viral spillover at species interfaces. Recurrent exposures from marine wildlife, tissue and cellular susceptibility to H5N1 IAVs, and connections to other IAV-susceptible terrestrial mammals establish the prerequisite conditions for vampire bats to spread IAVs between marine and terrestrial environments or to form a novel reservoir of highly pathogenic IAVs.
]]></description>
<dc:creator>Tu, I.-T.</dc:creator>
<dc:creator>Lynggaard, C.</dc:creator>
<dc:creator>Adams, L.</dc:creator>
<dc:creator>Walsh, S. K.</dc:creator>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Raveendran, S.</dc:creator>
<dc:creator>Turnbull, M. L.</dc:creator>
<dc:creator>Griffiths, M. E.</dc:creator>
<dc:creator>Ribeiro, R.</dc:creator>
<dc:creator>Perez, J. G.</dc:creator>
<dc:creator>Bazan, W. V.</dc:creator>
<dc:creator>Tello, C.</dc:creator>
<dc:creator>Zariquiey, C.</dc:creator>
<dc:creator>Rodriguez, K. P.</dc:creator>
<dc:creator>Risco, M.</dc:creator>
<dc:creator>Mamani, I. Q.</dc:creator>
<dc:creator>Chavez, W.</dc:creator>
<dc:creator>Villafuerte, R. Z.</dc:creator>
<dc:creator>Manuttupa, J. C.</dc:creator>
<dc:creator>Castro Namuche, J. P.</dc:creator>
<dc:creator>Moreira-Soto, A.</dc:creator>
<dc:creator>Drexler, J. F.</dc:creator>
<dc:creator>Delhon, G.</dc:creator>
<dc:creator>Faust, C.</dc:creator>
<dc:creator>Cardenas-Alayza, S.</dc:creator>
<dc:creator>Hutchinson, E.</dc:creator>
<dc:creator>Murcia, P. R.</dc:creator>
<dc:creator>Palmarini, M.</dc:creator>
<dc:creator>Bohmann, K.</dc:creator>
<dc:creator>Harvey, R.</dc:creator>
<dc:creator>Streicker, D. G.</dc:creator>
<dc:date>2025-11-11</dc:date>
<dc:identifier>doi:10.1101/2025.11.09.686930</dc:identifier>
<dc:title><![CDATA[Spillover of H5 influenza viruses to vampire bats at the marine-terrestrial interface]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.10.687569v1?rss=1">
<title>
<![CDATA[
Inhibiting epithelial cell extrusion and inflammation synergistically reverses asthmatic airway remodelling 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.10.687569v1?rss=1"
</link>
<description><![CDATA[
Asthma is a prevalent inflammatory disease marked by life-threatening airway constriction. Current therapies alleviate symptoms by relaxing airway smooth muscle and reducing inflammation but fail to address the underlying airway remodelling, which drives hyper-responsiveness and lung function decline. We previously showed that bronchoconstriction mechanics trigger pathological cell extrusion, wounding the epithelial barrier and perpetuating inflammation. Here, we reveal that a vicious cycle of epithelial damage and inflammation, perpetuates airways in a chronically wounded state. By inhibiting both extrusion and inflammation in mice with established airway asthma symptoms, we synergistically reverse airway remodelling and hyperresponsiveness to methacholine challenge. Moreover, this dual treatment reverts asthmatic transcriptomic and proteomic profiles to healthy states, restoring normal airway function. Our findings offer a novel therapeutic approach to not only halt but reverse asthma progression.
]]></description>
<dc:creator>Bagley, D. C.</dc:creator>
<dc:creator>Russell, T.</dc:creator>
<dc:creator>Martinez-Nunez, R. T.</dc:creator>
<dc:creator>Marcotti, S.</dc:creator>
<dc:creator>Rosenblatt, J.</dc:creator>
<dc:date>2025-11-11</dc:date>
<dc:identifier>doi:10.1101/2025.11.10.687569</dc:identifier>
<dc:title><![CDATA[Inhibiting epithelial cell extrusion and inflammation synergistically reverses asthmatic airway remodelling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.06.686978v1?rss=1">
<title>
<![CDATA[
Topological defects drive influenza glycoprotein lattice assembly on spherical membranes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.06.686978v1?rss=1"
</link>
<description><![CDATA[
Lipid-enveloped viruses, such as influenza virus, assemble by budding from infected cell membranes, packaging internal components including the genome and acquiring an envelope containing surface glycoproteins in the process. Influenza C virus possesses a single surface glycoprotein, the haemagglutinin-esterase-fusion (HEF) protein that forms hexagonal arrays1,2 on the membrane envelope and is sufficient for budding of spherical particles3. However, a two-dimensional hexagonal lattice cannot completely cover a spherical virus membrane without defects. Using electron cryotomography (cryo-ET), we study the structural arrangement of the influenza C virus surface and find the hexagonal HEF lattice contains 5-fold and 7-fold defects organised in grain boundaries. The number of excess dislocations increases with system size while maintaining a net topological charge near 12. Our observations of defects in spherical crystals on influenza C virus particles of varying radius and shape matches theoretical predictions of continuum elastic theory4 for the proliferation of defects on soft lattices and experimental observations5 on colloidal systems. These findings provide new principles for assembly of pleomorphic viruses, extending the description of defects required for viral lattice assembly beyond the Caspar-Klug theory6 developed for isometric viruses. Our study informs a wide range of molecular self-assembly processes in biology and may also have implications for developing lattice materials with curved surfaces.
]]></description>
<dc:creator>Liu, Z. B.</dc:creator>
<dc:creator>Halldorsson, S.</dc:creator>
<dc:creator>Calcraft, T.</dc:creator>
<dc:creator>Calder, L. J.</dc:creator>
<dc:creator>Rosenthal, P. B.</dc:creator>
<dc:date>2025-11-07</dc:date>
<dc:identifier>doi:10.1101/2025.11.06.686978</dc:identifier>
<dc:title><![CDATA[Topological defects drive influenza glycoprotein lattice assembly on spherical membranes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.06.686945v1?rss=1">
<title>
<![CDATA[
Adaptation of eukaryotic membrane homeostasis to species-specific cellular lipid landscapes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.06.686945v1?rss=1"
</link>
<description><![CDATA[
The physicochemical properties of biological membranes must be maintained within a range compatible with cellular physiology. In the face of external perturbations, membrane lipid homeostasis mechanisms sense and control membrane features. How such mechanisms evolve to function in organisms with different cellular lipid make-up is unknown. Here, we address this fundamental question by exploiting the natural divergence in membrane lipid composition between the related fission yeasts S. pombe and S. japonicus. Using lipidomics and transcriptomics, we show that the activity of the membrane-bound transcriptional activator Mga2, which regulates the {Delta}-9 desaturase Ole1 expression, is set to sense distinct levels of membrane unsaturation in the two species. Through retro-engineering and physiological experiments, we identify an evolutionary divergent but functionally constrained element within the juxtamembrane region of Mga2, which fine-tunes its performance to species-specific lipid composition. Our experiments indicate that high baseline expression of ole1, set by cis-regulatory elements in its upstream non-coding region, has redefined the dynamic range of Mga2 activation in S. pombe, supporting high lipidome unsaturation. Our work explores an "experiment of nature" to highlight the broad principles underlying the organisation and evolution of membrane homeostasis, which should be applicable to other genetic networks supporting cellular homeostatic processes.
]]></description>
<dc:creator>Gomez-Gil, E.</dc:creator>
<dc:creator>Guerra, P.</dc:creator>
<dc:creator>Gu, Y.</dc:creator>
<dc:creator>Foo, S.</dc:creator>
<dc:creator>Billard, B.</dc:creator>
<dc:creator>Balogh, G.</dc:creator>
<dc:creator>Peter, M.</dc:creator>
<dc:creator>Oliferenko, S.</dc:creator>
<dc:date>2025-11-07</dc:date>
<dc:identifier>doi:10.1101/2025.11.06.686945</dc:identifier>
<dc:title><![CDATA[Adaptation of eukaryotic membrane homeostasis to species-specific cellular lipid landscapes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.06.685569v1?rss=1">
<title>
<![CDATA[
A structural and regulatory framework for Atg9-containing vesicle formation and their Atg1-dependent remodelling during autophagy initiation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.06.685569v1?rss=1"
</link>
<description><![CDATA[
Autophagy is a complex intracellular degradation pathway that depends on the coordinated interplay between the core autophagy machinery and diverse membrane sources to drive the de novo formation of double-membrane vesicles, known as autophagosomes. Golgi-derived Atg9-containing vesicles are essential for this process, delivering membranes to the pre-autophagosomal structure (PAS). These vesicles contain the transmembrane proteins Atg9 and Atg27 and the peripheral membrane protein Atg23; however, the nature, function, and regulation of their interactions remain poorly understood.

Here, we systematically dissect the molecular interactions between Atg9, Atg23 and Atg27, and uncover their regulation in space and time. The bipartite binding mode by which Atg23 engages Atg9 provides a structural model for how Atg23 promotes vesicle budding. Furthermore, Atg1-dependent phosphorylation of Atg9 remodels its interactions with Atg23 and Atg27 at the PAS to support autophagy initiation. Together, these findings establish a molecular and regulatory framework for the earliest steps of autophagy.
]]></description>
<dc:creator>Oeo-Santos, C.</dc:creator>
<dc:creator>Knüpling, E.</dc:creator>
<dc:creator>Davis, C.</dc:creator>
<dc:creator>Meng, X.</dc:creator>
<dc:creator>Maslen, S.</dc:creator>
<dc:creator>Kunzelmann, S.</dc:creator>
<dc:creator>D'Antuono, R.</dc:creator>
<dc:creator>Olerinyova, A.</dc:creator>
<dc:creator>Auchynnikava, T.</dc:creator>
<dc:creator>Skehel, M.</dc:creator>
<dc:creator>Schreiber, A.</dc:creator>
<dc:date>2025-11-06</dc:date>
<dc:identifier>doi:10.1101/2025.11.06.685569</dc:identifier>
<dc:title><![CDATA[A structural and regulatory framework for Atg9-containing vesicle formation and their Atg1-dependent remodelling during autophagy initiation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.14.688404v1?rss=1">
<title>
<![CDATA[
PMGen: From Peptide-MHC Prediction to Neoantigen Generation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.14.688404v1?rss=1"
</link>
<description><![CDATA[
Accurate structural modeling of peptide-MHC (pMHC) complexes is a prerequisite for understanding adaptive immunity and developing data-driven immunotherapies. However, current tools are often limited by narrow class coverage, restricted peptide lengths, or insufficient accuracy for downstream design tasks. Here, we introduce PMGen (Peptide MHC Generator), an integrated framework for structure prediction and structure-guided design of variable-length peptides across MHC class I and II. By introducing Initial Guess and Template Engineering as strategies to enforce anchor constraints in AlphaFold2, PMGen achieves state-of-the-art structural fidelity with median peptide core RMSDs of 0.54 [A] for MHC-I and 0.33 [A] for MHC-II, outperforming five state-of-the-art methods. We further demonstrate that PMGen captures the subtle structural impact of single-point neoantigen mutations and that model confidence (pLDDT) reliably correlates with structural accuracy. We investigated two potential applications of our framework: structure-aware peptide design and generating data for machine learning (ML) models. To this end, we introduced a framework to sample peptides with preserved structures and improved binding affinity. As an example for ML application, we fine-tuned ProteinMPNN on PMGen-modeled structures. This improved sequence recovery from 0.19 to 0.40 compared to the baseline. Ultimately, PMGen bridges the gap between high-fidelity structural prediction and downstream sequence design, offering a scalable solution to generate the large-scale, high-quality structural datasets required to train advanced predictive models in immunology. Available at https://github.com/soedinglab/PMGen.
]]></description>
<dc:creator>Asgary, A. H.</dc:creator>
<dc:creator>Aleyasin, A.</dc:creator>
<dc:creator>Mehl, J. A.</dc:creator>
<dc:creator>Ludewig, B.</dc:creator>
<dc:creator>Mishto, M.</dc:creator>
<dc:creator>Liepe, J.</dc:creator>
<dc:creator>Soeding, J.</dc:creator>
<dc:date>2025-11-14</dc:date>
<dc:identifier>doi:10.1101/2025.11.14.688404</dc:identifier>
<dc:title><![CDATA[PMGen: From Peptide-MHC Prediction to Neoantigen Generation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.12.687981v1?rss=1">
<title>
<![CDATA[
Siglec-engaging immunosuppressive sialoglycans are upregulated in prostate cancer and are targetable to suppress bone metastasis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.12.687981v1?rss=1"
</link>
<description><![CDATA[
Prostate cancer is a leading cause of male cancer-related deaths over the age of 50. New treatment options for prostate cancer are urgently needed, especially for tumours that have spread to bone. Aberrant sialylation holds substantial potential for the discovery of new therapeutic targets but has remained relatively unexplored in the context of prostate cancer, primarily due to the lack of reliable reagents for detecting tumour sialoglycans in clinical tissue. Here, we address this knowledge gap using high-affinity Siglec-based sialoglycan-binding reagents (HYDRAs) to quantify tumour sialoglycans in tissues representing the full clinical heterogeneity of prostate tumours. Using HYDRA immunohistochemistry, we show that sialoglycans that can engage Siglec-3, -7, and -9 are upregulated in primary prostate cancer tissue and sialoglycan ligands for Siglec-7 correlate with prostate cancer bone metastasis and poorer patient prognosis. Analysis of prostate-derived tumours growing in bone reveals Siglec receptors are expressed by immune cells in the bone metastatic tumour microenvironment, suggesting that this axis may play a role in immune cell functions in bone metastatic prostate cancer. Indicating this is clinically actionable, an engineered bisialidase (E-612) can effectively strip Siglec ligands from prostate cancer cells and prolong survival times of mice with bone metastasis. Our findings identify a novel mechanism involving Siglec-engaging sialoglycans in driving the growth of prostate cancer bone metastasis and demonstrate how this axis can be targeted to impede lethal prostate cancer progression.
]]></description>
<dc:creator>Peng, Z.</dc:creator>
<dc:creator>Hodgson, K.</dc:creator>
<dc:creator>Fisher, M.</dc:creator>
<dc:creator>Mei, S.</dc:creator>
<dc:creator>Orozco-Moreno, M.</dc:creator>
<dc:creator>Cao, L.</dc:creator>
<dc:creator>Kemp, M.</dc:creator>
<dc:creator>Gatlin, W.</dc:creator>
<dc:creator>Donald, L.</dc:creator>
<dc:creator>Zeng, F.</dc:creator>
<dc:creator>Lawson, M. A.</dc:creator>
<dc:creator>Ungar, D.</dc:creator>
<dc:creator>Sykes, D. B.</dc:creator>
<dc:creator>Elliott, D. J.</dc:creator>
<dc:creator>Peng, L.</dc:creator>
<dc:creator>Schumann, B.</dc:creator>
<dc:creator>Wang, N.</dc:creator>
<dc:creator>Munkley, J.</dc:creator>
<dc:date>2025-11-13</dc:date>
<dc:identifier>doi:10.1101/2025.11.12.687981</dc:identifier>
<dc:title><![CDATA[Siglec-engaging immunosuppressive sialoglycans are upregulated in prostate cancer and are targetable to suppress bone metastasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.11.682326v1?rss=1">
<title>
<![CDATA[
Pathogenic Keratinocyte States and Fibroblast Niches Define the Tissue Microenvironment in Severe Hidradenitis Suppurativa 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.11.682326v1?rss=1"
</link>
<description><![CDATA[
Hidradenitis suppurativa (HS) is a chronic inflammatory skin disease characterized by recurrent painful abscesses and tunnels in flexural sites. The mechanisms driving HS pathogenesis, particularly the roles of keratinocytes and fibroblasts in the HS inflammatory ecosystem, remain poorly understood. To characterise the cellular and molecular landscape of HS, we analyzed lesional skin from severe HS patients using single-cell RNA-sequencing and spatial transcriptomics to identify key keratinocyte states and cellular interactions, with a focus on fibroblast-keratinocyte crosstalk. Our study identifies a novel migratory S100+ pathogenic keratinocyte state and highlights critical interactions between COL6A5+ fibroblasts in HS lesion formation. We also detect tertiary lymphoid organ (TLO)-like structures enriched with activated B and plasma cells interacting with APOD+ fibroblasts, implicating their role in HS chronic inflammation. We show that fibroblast interactions with HS keratinocytes and specific immune cells, such as Langerhans cells, are key drivers of HS pathogenesis, and that two main fibroblast niches are present within the HS microenvironment. Targeting these cellular networks may offer new therapeutic strategies for HS management, and highlight the potential limitations of targeting individual pathways in isolation, when treating pathology present in HS tissue.
]]></description>
<dc:creator>Du-Harpur, X.</dc:creator>
<dc:creator>Ganier, C.</dc:creator>
<dc:creator>Mazin, P.</dc:creator>
<dc:creator>Cheshire, C.</dc:creator>
<dc:creator>Rashidghamat, E.</dc:creator>
<dc:creator>Luscombe, N. M.</dc:creator>
<dc:creator>Lynch, M. D.</dc:creator>
<dc:creator>Watt, F. M.</dc:creator>
<dc:date>2025-11-12</dc:date>
<dc:identifier>doi:10.1101/2025.11.11.682326</dc:identifier>
<dc:title><![CDATA[Pathogenic Keratinocyte States and Fibroblast Niches Define the Tissue Microenvironment in Severe Hidradenitis Suppurativa]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.11.687613v1?rss=1">
<title>
<![CDATA[
WNT inhibition primes the transcriptional landscape of mesoderm to initiate a phased ventricular cardiomyocyte specification programme 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.11.687613v1?rss=1"
</link>
<description><![CDATA[
Background and AimsCardiac reprogramming holds promise for treating ischemic heart diseases and advancing personalised medicine, but current approaches do not yet reliably generate human functional, homogeneous cardiomyocyte cultures. This limited success likely reflects our incomplete understanding of hierarchical transcriptional programmes guiding cardiomyocyte specification. Here, we modelled cardiomyocyte differentiation aiming to uncover gene regulatory networks (GRNs) guiding early human ventricular cardiomyocyte development. Our study focused on defining how inhibition of WNT signalling remodels the transcriptional landscape underlying human cardiomyocyte differentiation, since both in vivo cardiac development and in vitro cardiomyocyte differentiation protocols require WNT inhibition.

MethodsWe modelled left ventricular cardiomyocyte differentiation from human pluripotent stem cells and experimentally manipulated WNT signalling to uncover transcriptional responses using single-cell RNA sequencing. Bioinformatics analysis defined cell identities, reconstructed differentiation trajectories, and inferred WNT inhibition-dependent gene expression and regulatory networks driving cardiomyocyte specification.

ResultsWe found that WNT inhibition (WNTi) decisively redirects mesoderm cells towards a cardiomyocyte progenitor fate, expanding their numbers while limiting alternative trajectories. GRN inference revealed both WNTi-dependent and -independent programmes and a hierarchical cascade of transcription factors driving the mesoderm-to-cardiomyocyte progenitor transition. Notably, MEIS2 emerged as a central WNTi-independent regulator, while WNTi-responsive networks featured early (ISL1, PBX3, TBX5 and KLF1) and late (MEF2-related genes, GATA-related genes, PBX1, CREM, FOXP1 and NKX3-1) factors. Mesodermal GRNs were primed for cardiomyocyte specification in WNTi-treated cultures but, in the absence of WNTi, mesodermal GRNs remained ambiguous, activating ISL1 and PBX3 but failing to establish cardiomyocyte commitment and subsequent differentiation into contractile cells.

ConclusionThis work provides the first comprehensive dissection of WNTi-dependent and -independent regulatory hierarchies guiding human ventricular cardiomyocyte specification and highlights new transcriptional players which could improve cardiac reprogramming efficiency and fidelity.

Translational PerspectiveDeciphering the transcriptional programmes that drive early cardiomyocyte specification has clear translational potential for regenerative therapies and cardiac reprogramming. By modelling left ventricular cardiomyocyte differentiation from hPSCs, our study highlights how inhibition of WNT signalling promotes ventricular cardiomyocyte progenitor commitment while restricting alternative fates. The discovery of both WNTi-dependent and -independent transcriptional activators, including some not previously linked to cardiomyocyte development, provides new insights for improving the efficiency and fidelity of direct cardiac reprogramming. Importantly, these TF candidates may also guide the development of targeted therapies for ischemic heart disease and inform personalised approaches for repairing and regenerating damaged myocardium.
]]></description>
<dc:creator>Velecela, V.</dc:creator>
<dc:creator>Bassil, A.</dc:creator>
<dc:creator>Fawcett, E.</dc:creator>
<dc:creator>Smith, E.</dc:creator>
<dc:creator>Konstantopoulos, D.</dc:creator>
<dc:creator>Salmen, F.</dc:creator>
<dc:creator>Bernardo, A. S.</dc:creator>
<dc:creator>Hoppler, S. S.</dc:creator>
<dc:date>2025-11-12</dc:date>
<dc:identifier>doi:10.1101/2025.11.11.687613</dc:identifier>
<dc:title><![CDATA[WNT inhibition primes the transcriptional landscape of mesoderm to initiate a phased ventricular cardiomyocyte specification programme]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.18.689114v1?rss=1">
<title>
<![CDATA[
PETRA: Prime editing of transcribed regulatory elements to assay expression 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.18.689114v1?rss=1"
</link>
<description><![CDATA[
Predicting how changes in human DNA sequence impact gene expression remains challenging. Here, we present PETRA (Prime Editing of Transcribed Regulatory elements to Assay expression), a multiplexed genome editing method to quantify the effects of regulatory variants at scale. PETRA leverages the delivery of variants to abundantly transcribed regions of genes such that sequence-specific effects on mRNA expression can be read out by amplicon sequencing. We demonstrate PETRA in Jurkat cells by scoring 13,935 six-nucleotide insertions delivered to the 5 untranslated regions (5 UTRs) of four genes important for T cell responses, namely VAV1, IL2RA, CD28 and OTUD7B. Effects on expression are linked to the creation of new transcription factor binding sites (TFBSs), as well as to alterations in splicing and translation initiation. Combinatorial delivery of TFBSs identified using PETRA generates alleles that increase mRNA expression more than 10-fold. Additionally, we extend PETRA to primary human T cells to compare effects across cell types and use our data to assess the performance of computational models. These results establish PETRA as a flexible means of dissecting and reprogramming the logic of gene regulation across genomic contexts and cell types.
]]></description>
<dc:creator>Reyes, M. A.</dc:creator>
<dc:creator>Herger, M.</dc:creator>
<dc:creator>Cubitt, L.</dc:creator>
<dc:creator>Findlay, G. M.</dc:creator>
<dc:date>2025-11-18</dc:date>
<dc:identifier>doi:10.1101/2025.11.18.689114</dc:identifier>
<dc:title><![CDATA[PETRA: Prime editing of transcribed regulatory elements to assay expression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.18.689008v1?rss=1">
<title>
<![CDATA[
Human airway epithelia clear rhinovirus-infected cells using two waves of cell extrusion 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.18.689008v1?rss=1"
</link>
<description><![CDATA[
Epithelial barriers represent the first line of defence against pathogens, yet their role in innate immunity is typ-ically relegated to pathogen detection and immune cell recruitment. This perspective ignores a fundamental evolutionary principle: epithelia defended against pathogens long before complex immune systems evolved. Here, we demonstrate that human bronchial epithelial monolayers retain this ancestral capacity, autono-mously clearing rhinovirus (RV) within 24 hours by selectively extruding infected cells--a process we term virus-induced cell extrusion (VICE). VICE occurs in two waves: a rapid response occurring independently of virus entry, followed by a replication-dependent wave. Barrier-defective epithelia that cannot extrude fail to clear RV. While extrusion maintains barrier integrity and eliminates local infection, it also expels virus-laden cells, promoting transmission. Thus, VICE enables leukocyte-independent epithelial defence while inadver-tently promoting viral transmission, reflecting an evolutionary strategy that prioritizes barrier integrity over containment. These findings redefine epithelia as central players in viral pathogenesis and host protection.
]]></description>
<dc:creator>Fore, F.</dc:creator>
<dc:creator>Bagley, D. C.</dc:creator>
<dc:creator>Martinez-Nunez, R. T.</dc:creator>
<dc:creator>Aniscenko, J.</dc:creator>
<dc:creator>Johnston, S.</dc:creator>
<dc:creator>Rosenblatt, J.</dc:creator>
<dc:date>2025-11-18</dc:date>
<dc:identifier>doi:10.1101/2025.11.18.689008</dc:identifier>
<dc:title><![CDATA[Human airway epithelia clear rhinovirus-infected cells using two waves of cell extrusion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.21.689802v1?rss=1">
<title>
<![CDATA[
A divergent Plasmodium NEK4 acts as a key regulator driving the early events of meiosis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.21.689802v1?rss=1"
</link>
<description><![CDATA[
Meiosis is a conserved yet evolutionarily varied process underpinning sexual reproduction in eukaryotes. In the malaria parasite Plasmodium, meiosis is unconventional: it occurs immediately after fertilisation (post-zygotic) and must be coordinated with the transformation of the zygote into a motile ookinete. The mechanisms synchronising these meiotic and morphogenetic programmes remain unknown. Here, we identify the Plasmodium berghei NIMA-related kinase NEK4 as a key regulator that couples meiotic initiation with zygote morphogenesis. Using ultrastructure expansion microscopy, we show that NEK4 accumulates at the microtubule-organising centre (MTOC) and the apical polar complex (APC) shortly after fertilisation, preceding the assembly of perinuclear and cortical microtubules. We reveal that Plasmodium zygotes undergo MTOC-associated nuclear migration, analogous to the meiotic nuclear movement in fission yeast. Deletion of the Pbnek4 gene results in complete developmental arrest: MTOC duplication and microtubule formation are blocked, chromatin remains uncondensed, and nuclear migration and cell polarity fail to establish. Transcriptomic and phosphoproteomic analyses reveal that absence of NEK4 causes a collapse in transcriptional and phosphoregulatory networks governing meiosis and cytoskeletal organisation, leading to reduced expression and phosphorylation of important players, including HOP1, REC8, and AP2-O. These findings establish NEK4 as a key regulator driving meiotic entry and zygote maturation.
]]></description>
<dc:creator>Yanase, R.</dc:creator>
<dc:creator>Hair, M.</dc:creator>
<dc:creator>Zeeshan, M.</dc:creator>
<dc:creator>Ferguson, D. J. P.</dc:creator>
<dc:creator>Brady, D.</dc:creator>
<dc:creator>Pasquarello, C.</dc:creator>
<dc:creator>Bottrill, A.</dc:creator>
<dc:creator>Bhanvadia, S.</dc:creator>
<dc:creator>Neal, A.</dc:creator>
<dc:creator>Tromer, E. C.</dc:creator>
<dc:creator>Le Roch, K. G.</dc:creator>
<dc:creator>Hainard, A.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:creator>Vaughan, S.</dc:creator>
<dc:creator>Guttery, D. S.</dc:creator>
<dc:creator>Tewari, R.</dc:creator>
<dc:date>2025-11-23</dc:date>
<dc:identifier>doi:10.1101/2025.11.21.689802</dc:identifier>
<dc:title><![CDATA[A divergent Plasmodium NEK4 acts as a key regulator driving the early events of meiosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.20.689507v1?rss=1">
<title>
<![CDATA[
Rv3400 is a phosphoglucomutase required for trehalose metabolism in Mycobacterium tuberculosis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.20.689507v1?rss=1"
</link>
<description><![CDATA[
Mycobacterium tuberculosis is a global killer causing over a million deaths from tuberculosis (TB) every year. It is therefore a major burden on human health. To reduce the deadly impact of TB, we need a better understanding of the strategies used by M. tuberculosis to adapt its metabolism to survive and persist in the human host. This will help us design better strategies for TB treatment and control. Previous enzymological studies have reported the Mtb rv3400 gene as encoding a {beta}-phosphoglucomutase; however, its role in M. tuberculosis metabolism was not investigated. In the present study, we show that deletion of rv3400 leads to a 30-fold increase in {beta}-D-glucose 1-phosphate, confirming its primary function as a {beta}-phosphoglucomutase. Additionally, deletion of rv3400 leads to a growth defect when trehalose is the sole carbon source indicting that this enzyme is required for optimal use of trehalose, an essential disaccharide in mycobacteria.

ImportanceTrehalose metabolism plays a corner stone in Mycobacterium tuberculosis physiology and virulence. Hence, a better understanding of the metabolism of this essential disaccharide is required to develop novel strategies to eradicate Tuberculosis. Here we report on the characterisation of the strain lacking a {beta}-phosphoglucomutase, encoded by the gene rv3400. We show that deletion of rv3400 leads to an increase of {beta}-D-glucose 1-phosphate and a growth defect when using trehalose as sole carbon source. Taken together, the data presented here provide evidence that Rv3400 is required for the catabolism of trehalose.
]]></description>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Ruecker, N.</dc:creator>
<dc:creator>Faulkner, V.</dc:creator>
<dc:creator>Rachineni, K.</dc:creator>
<dc:creator>Al-Saffar, N.</dc:creator>
<dc:creator>VillacampaTeixeira, P.</dc:creator>
<dc:creator>Dias da Costa, T.</dc:creator>
<dc:creator>Johnson, E.</dc:creator>
<dc:creator>Robertson, B. D.</dc:creator>
<dc:creator>Ehrt, S.</dc:creator>
<dc:creator>Larrouy-Maumus, G.</dc:creator>
<dc:date>2025-11-20</dc:date>
<dc:identifier>doi:10.1101/2025.11.20.689507</dc:identifier>
<dc:title><![CDATA[Rv3400 is a phosphoglucomutase required for trehalose metabolism in Mycobacterium tuberculosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.19.689206v1?rss=1">
<title>
<![CDATA[
XIST Drives X-Chromosome Inactivation and Safeguards Female Extraembryonic Cells in Humans 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.19.689206v1?rss=1"
</link>
<description><![CDATA[
Dosage compensation of sex chromosomes through X-chromosome inactivation (XCI) is required for mice extra-embryonic tissue growth and embryo development. The species specificity in mechanisms and timing leading to XCI during early embryogenesis, however, left the key question of the interdependence between XCI and human development open. Here, we show that the differentiation of naive human pluripotent stem cells to trophoblast stem cells and extraembryonic mesoderm cells triggers XCI. The inactive X chromosome, however, displays an atypical chromatin state, lacking classical enrichment of heterochromatin markers and DNA methylation. We demonstrate that extraembryonic differentiation and XCI are kinetically and functionally linked. Using loss of function approaches, we prove that XIST is required for human XCI establishment. We also reveal that XCI is key for the survival of human female extraembryonic cells. Our work therefore links XCI to the formation of extraembryonic annexes, with important consequences for human reproductive biology.

HIGHLIGHTSO_LINaive hPSCs to EXMCs and TSCs differentiation recapitulates human XCI
C_LIO_LIThe Xi has an unusual chromatin status in human extraembryonic cells
C_LIO_LIXIST is required for the establishment of human XCI
C_LIO_LIXCI supports healthy development of human female extraembryonic cells
C_LI
]]></description>
<dc:creator>Panda, A.</dc:creator>
<dc:creator>Carrillo, L.</dc:creator>
<dc:creator>Balaton, B. P.</dc:creator>
<dc:creator>Brouillet, J.</dc:creator>
<dc:creator>Nshemereirwe, S.</dc:creator>
<dc:creator>Bonroy, J.</dc:creator>
<dc:creator>Alfeghaly, C.</dc:creator>
<dc:creator>Facchinello, R.</dc:creator>
<dc:creator>Khodeer, S.</dc:creator>
<dc:creator>Boers, R.</dc:creator>
<dc:creator>Castel, G.</dc:creator>
<dc:creator>London, C.</dc:creator>
<dc:creator>Cazottes, E.</dc:creator>
<dc:creator>Moscatelli, M.</dc:creator>
<dc:creator>Tchinda, R. S.</dc:creator>
<dc:creator>Pham, T. X. A.</dc:creator>
<dc:creator>To, S. K.</dc:creator>
<dc:creator>Allsop, R. N.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Staneva, D.</dc:creator>
<dc:creator>Rugg-Gunn, P.</dc:creator>
<dc:creator>Niakan, K.</dc:creator>
<dc:creator>Gribnau, J.</dc:creator>
<dc:creator>Ouimette, J.-F.</dc:creator>
<dc:creator>Rougeulle, C.</dc:creator>
<dc:creator>Pasque, V.</dc:creator>
<dc:date>2025-11-19</dc:date>
<dc:identifier>doi:10.1101/2025.11.19.689206</dc:identifier>
<dc:title><![CDATA[XIST Drives X-Chromosome Inactivation and Safeguards Female Extraembryonic Cells in Humans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.25.690426v1?rss=1">
<title>
<![CDATA[
Oncogenic RAS activity is linked to immune priming and adenosine-driven immune evasion in lung adenocarcinoma 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.25.690426v1?rss=1"
</link>
<description><![CDATA[
Lung adenocarcinoma (LUAD) is a leading cause of cancer death worldwide, with RAS signalling as a key oncogenic driver. Although KRAS mutations have been linked to immune evasion in preclinical models, the relationship between RAS activity and tumour immunity or response to immunotherapy in patients remains unclear. Here, we applied our previously validated RAS84 transcriptional signature to stratify LUAD patient cohorts and dissect the immune landscape associated with RAS signalling. We report that tumours with elevated RAS activity exhibited features of immune priming, including increased immune infiltration, interferon response, and immune checkpoint gene expression, and showed improved progression-free survival in an independent cohort of patients treated with anti-PD-1. Yet, in both LUAD tumours and cell lines, RAS activity also correlated with elevated immunosuppressive interstitial adenosine mediated by transcriptional regulation of several components of the adenosinergic pathway. In orthotopic pre-clinical models of high-RAS activity lung tumours, blocking adenosine signalling delayed tumour growth and improved response to anti-PD-1 and KRAS inhibition, with a significant effect on innate immunity. This study reveals a dual role for RAS signalling in tumour progression, fostering a pro-immunogenic environment whilst simultaneously dampening anti-tumoural immunity via mechanisms including extracellular adenosine accumulation. Stratifying patients based on RAS transcriptional activity, rather than genetic alterations alone, could inform immunotherapy strategies and improve clinical outcomes.
]]></description>
<dc:creator>de Carne Trecesson, S.</dc:creator>
<dc:creator>East, P.</dc:creator>
<dc:creator>Pillsbury, C.</dc:creator>
<dc:creator>Silva dos Santos, M.</dc:creator>
<dc:creator>Cha, H.</dc:creator>
<dc:creator>Colliver, E.</dc:creator>
<dc:creator>Gilmore, T.</dc:creator>
<dc:creator>Rana, S.</dc:creator>
<dc:creator>Moore, C.</dc:creator>
<dc:creator>Lighterness, S.</dc:creator>
<dc:creator>Caswell, D. R.</dc:creator>
<dc:creator>Boumelha, J.</dc:creator>
<dc:creator>Tomaschko, M.</dc:creator>
<dc:creator>Baer, R.</dc:creator>
<dc:creator>Eyles, J.</dc:creator>
<dc:creator>Teixeira, B.</dc:creator>
<dc:creator>Saeed, M.</dc:creator>
<dc:creator>Litchfield, K.</dc:creator>
<dc:creator>Molina-Arcas, M.</dc:creator>
<dc:creator>Lee, S.-H.</dc:creator>
<dc:creator>MacRae, J.</dc:creator>
<dc:creator>Hobson, P.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Downward, J.</dc:creator>
<dc:date>2025-11-26</dc:date>
<dc:identifier>doi:10.1101/2025.11.25.690426</dc:identifier>
<dc:title><![CDATA[Oncogenic RAS activity is linked to immune priming and adenosine-driven immune evasion in lung adenocarcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.12.01.691495v1?rss=1">
<title>
<![CDATA[
SPIN90 modulates the architecture of lamellipodial actin in an ARPC5L dependent fashion. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.12.01.691495v1?rss=1"
</link>
<description><![CDATA[
When stimulated by nucleation-promoting factors such as WAVE, the Arp2/3 complex generates branched actin networks. In contrast, when activated by SPIN90, the Arp2/3 complex generates linear actin filaments. The Arp2/3 complex in mammals, consists of 8 iso-complexes with dieerent properties as there are two isoforms of Arp3, ArpC1 and ArpC5. Here, using recombinant Arp2/3 iso-complexes with defined compositions, we show that SPIN90 selectively activates ArpC5L- rather than ArpC5- containing complexes to generate linear actin filaments. Consistent with this, SPIN90 at the leading edge of migrating cells enhances the recruitment of ArpC5L-, but not ArpC5- containing, Arp2/3 complexes to lamellipodia. These SPIN90-Arp2/3-ArpC5L complexes generate linear actin filaments that integrate into lamellipodia and impact protrusion eeiciency. Moreover, using polarised light microscopy, we show that loss of SPIN90 leads to an enrichment in actin filaments oriented more perpendicular to the plasma membrane. Our results demonstrate that SPIN90 regulates the architecture and dynamics of lamellipodial actin in an Arp2/3 iso-complex dependent fashion.

SummarySPIN90 selectively recruits and activates ArpC5L containing Arp2/3 to shape the architecture and dynamics of lamellipodial actin.
]]></description>
<dc:creator>Cao, L.</dc:creator>
<dc:creator>Basant, A.</dc:creator>
<dc:creator>Mladenov, M.</dc:creator>
<dc:creator>Kogata, N.</dc:creator>
<dc:creator>Jegou, A.</dc:creator>
<dc:creator>Romet-Lemonne, G.</dc:creator>
<dc:creator>Brasselet, S.</dc:creator>
<dc:creator>Mavrakis, M.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:date>2025-12-02</dc:date>
<dc:identifier>doi:10.64898/2025.12.01.691495</dc:identifier>
<dc:title><![CDATA[SPIN90 modulates the architecture of lamellipodial actin in an ARPC5L dependent fashion.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.28.691009v1?rss=1">
<title>
<![CDATA[
The ERCC6L2-MRI-KU complex coordinates NHEJ at staggered DNA double-strand breaks. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.28.691009v1?rss=1"
</link>
<description><![CDATA[
ERCC6L2 disease is a recessive bone marrow failure (BMF) syndrome caused by mutations in the SNF2-like putative DNA helicase ERCC6L2. While implicated in DNA replication, double strand break (DSB) repair via non-homologous end joining (NHEJ), and interstrand crosslink (ICL) repair, how ERCC6L2 supports haematopoietic longevity remains unclear. Investigating this in vivo, we find that an Ercc6l2-deficient haematopoietic stem and progenitor cell (HSPC) compartment in mice is unexpectedly resilient. Ercc6l2 loss was also tolerated in mice co-deficient for endogenous formaldehyde detoxification, which precipitates early-onset BMF in models of Fanconi anaemia. Instead, Ercc6l2-deficient mice display a mild immunodeficiency, arising from defects in immunoglobulin class-switch recombination (CSR), that synergise with shieldin-deficiency, implicating ERCC6L2 and shieldin in distinct repair mechanisms. Furthermore, we demonstrate that ERRC6L2 stimulates chromosome fusions in the context of staggered, but not blunt dysfunctional telomeres. We reconcile ERCC6L2s NHEJ function through proteomic elucidation of its endogenous interactome and AlphaFold structural modelling to reveal a complex formed of ERCC6L2 and KU that is bridged by the NHEJ accessory factor MRI/CYREN. Consequently, ERCC6L2-MRI inter-dependence characterises CSR. Together, our findings implicate the ERCC6L2-MRI complex as a KU-regulatory DNA translocase coordinating classical-NHEJ at staggered-end DSBs. We suggest that similar staggered-end breaks represent the pathological substrates driving haematopoietic failure in ERCC6L2 disease.
]]></description>
<dc:creator>Reichl, P. I.</dc:creator>
<dc:creator>Sonmez, C.</dc:creator>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>King, A.</dc:creator>
<dc:creator>Davies, B.</dc:creator>
<dc:creator>Wilkinson, A. C.</dc:creator>
<dc:creator>Lottersberger, F.</dc:creator>
<dc:creator>Chapman, J. R.</dc:creator>
<dc:date>2025-11-28</dc:date>
<dc:identifier>doi:10.1101/2025.11.28.691009</dc:identifier>
<dc:title><![CDATA[The ERCC6L2-MRI-KU complex coordinates NHEJ at staggered DNA double-strand breaks.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.12.10.693488v1?rss=1">
<title>
<![CDATA[
Tracking gene expression of single mitochondria in live neurons using nanotweezers 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.12.10.693488v1?rss=1"
</link>
<description><![CDATA[
Neurons are highly polarised cells that depend on mitochondria for energy and signalling homeostasis. Importantly, energy and signalling requirements vary considerably across individual neurons both spatially and temporally. Therefore, to fully understand neuronal mitochondria, methods are needed to analyse mitochondria in live cells over time. The nanotweezer, a minimally invasive single-cell sampling technique, enables precise extraction a individual mitochondria from defined subcellular locations. Here, we combine single-mitochondrial extraction from live neurons with mitochondrial gene expression tracking and mtDNA profiling. By tracking mitochondrial gene expression in the same neurons over time, we reveal a downregulation of mitochondrial genes MT-ND1 and MT-ATP6 following exposure to -synuclein aggregates, independent of the proximity of the aggregates to the sampled mitochondria. Our approach provides precise, dynamic measurements of mitochondrial composition and gene expression in vivo at single-organelle resolution, enabling mechanistic studies of neuronal mitochondrial heterogeneity and its perturbation in models of neurodegeneration.
]]></description>
<dc:creator>Sahota, A.</dc:creator>
<dc:creator>Nadappuram, B.</dc:creator>
<dc:creator>Allerton, S. C.</dc:creator>
<dc:creator>Lesept, F.</dc:creator>
<dc:creator>Howden, J.</dc:creator>
<dc:creator>Claxton, S.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Aprile, F. A.</dc:creator>
<dc:creator>Kittler, J.</dc:creator>
<dc:creator>Devine, M. J.</dc:creator>
<dc:creator>Edel, J.</dc:creator>
<dc:creator>Ivanov, A.</dc:creator>
<dc:date>2025-12-13</dc:date>
<dc:identifier>doi:10.64898/2025.12.10.693488</dc:identifier>
<dc:title><![CDATA[Tracking gene expression of single mitochondria in live neurons using nanotweezers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.12.11.693708v1?rss=1">
<title>
<![CDATA[
PD-L2 Regulates Natural Antibody and IL-10 Secretion by B-1 cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.12.11.693708v1?rss=1"
</link>
<description><![CDATA[
B-1 cells are innate-like lymphocytes that play a critical role in homeostasis by secreting natural antibodies, typically IgM, and immunosuppressive molecules such as IL-10. However, the regulation of these processes in B-1 cells remains poorly understood. Here, we demonstrate that PD-L2, a surface receptor expressed by all B-1 cells, regulates B-1 cell effector functions. We show that in contrast to the Mus musculus castaneus mouse and other mammals, commonly used laboratory mouse strains have a mutation resulting in a premature stop codon and hence a truncated PD-L2 lacking an intracellular domain. By reverting this mutation, we generated mice expressing full-length PD-L2 containing an intracellular domain. Using these, we found that PD-L2 signaling via the intracellular domain increases IL-10 secretion and decreases natural IgM secretion by B-1 cells, potentially due to reduced expression of plasma cell identity genes Blimp-1 and IRF4. These findings indicate direct cell-cell interaction-dependent regulation of B-1 cell effector functions mediated by PD-L2 signaling into B-1 cells.
]]></description>
<dc:creator>Amendt, T.</dc:creator>
<dc:creator>Vanes, L.</dc:creator>
<dc:creator>Tybulewicz, V.</dc:creator>
<dc:date>2025-12-14</dc:date>
<dc:identifier>doi:10.64898/2025.12.11.693708</dc:identifier>
<dc:title><![CDATA[PD-L2 Regulates Natural Antibody and IL-10 Secretion by B-1 cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.12.16.694688v1?rss=1">
<title>
<![CDATA[
Placental insufficiency causes fetal growth restriction in mice lacking Delta-like homologue 1 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.12.16.694688v1?rss=1"
</link>
<description><![CDATA[
Fetal growth restriction (FGR) affects between 3-7% of pregnancies, is associated with increased perinatal morbidity and mortality, and linked to failure of placental function. The placenta is the key transient organ in pregnancy that directs nutrient transfer, intermediary metabolism and the production of hormones that drive maternal metabolic adaptations essential for pregnancy and lactation. The exchange surface of the placenta is formed in early development by the interaction between trophoblast cells that enclose the maternal blood and extraembryonic mesodermal cells that comprise the fetal vasculature. Despite recent insights into trophoblast development derived from novel in-vitro approaches, the processes driving extraembryonic mesoderm development are not well explored. This is due to a dearth of studies employing unbiased approaches to interrogate extraembryonic mesoderm cell populations. Here we use genetic labelling techniques to separate molecular events occurring in the trophoblast from those in the mesodermal layers of the placenta. In combination with conditional targeting, we show that the imprinted gene Dlk1 is a key player in providing nutrients to the embryo by controlling the placental surface area available for nutrient exchange, and by modulating the production of placental hormones that promote maternal nutrient provision in pregnancy.
]]></description>
<dc:creator>Charalambous, M.</dc:creator>
<dc:creator>Vignola, M. L.</dc:creator>
<dc:creator>Esse, R.</dc:creator>
<dc:creator>Scagliotti, V.</dc:creator>
<dc:creator>Servadei, C.</dc:creator>
<dc:creator>Kardasz, D.</dc:creator>
<dc:creator>Marinelli, E.</dc:creator>
<dc:creator>Dent, C.</dc:creator>
<dc:date>2025-12-18</dc:date>
<dc:identifier>doi:10.64898/2025.12.16.694688</dc:identifier>
<dc:title><![CDATA[Placental insufficiency causes fetal growth restriction in mice lacking Delta-like homologue 1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.12.16.694631v1?rss=1">
<title>
<![CDATA[
Mesodermal-specific MECP2 expression in Drosophila induces visceral and skeletal muscle defects rescued by butyrate supplementation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.12.16.694631v1?rss=1"
</link>
<description><![CDATA[
BackgroundPatients affected by Rett syndrome (RTT) and MECP2 duplication syndrome (MDS) experience disabling muscle weakness and gastrointestinal dysmotility of unclear origin. Whether these defects arise cell-autonomously, rather than secondarily to neural dysfunction, and which developmental windows are most vulnerable to MeCP2 disfunction remains unresolved. MeCP2 is a dosage-sensitive transcriptional regulator, whose functions are tightly linked to chromatin states. Because short-chain fatty acids (SCFAs) are known to inhibit histone deacetylases (HDACs), a tractable in vivo model is needed to test the effect of HDAC modulation on muscle defects.

MethodsWe misexpressed human MECP2 in the Drosophila melanogaster mesoderm that gives rise to skeletal and visceral muscles. We analyzed quantitatively their morphology and function. To assess the effects of SCFA supplementation, we also supplemented diets with sodium butyrate (NaB), Lalbaay(R), a NaB-containing supplement, acetate (AcOH), and valproate (VPA).

FindingsMECP2 misexpression caused pre-eclosion lethality, thinning of larval skeletal fibers with nuclear mispositioning and altered mitochondria. Functionally, it reduced locomotion, decreased food transit and gut peristalsis. Phenotypes were strongest when expression began during development. NaB and VPA supplementation rescue most of these phenotypes, consistent with their histone-deacetylase (HDAC) activity. Defects were not observed upon comparable misexpression of an RTT-associated MeCP2 loss-of-function variant, indicating that they might be relevant to pathogenesis of MECP2-related disorders.

InterpretationOur genetic in vivo analysis models peripheral effects of MeCP2 dysregulation and their amelioration, supporting the possibility of HDAC-targeted strategies for MECP2-related muscle and gastrointestinal dysfunction.
]]></description>
<dc:creator>Consonni, G.</dc:creator>
<dc:creator>Kozluca, M.</dc:creator>
<dc:creator>Gagliani, M. C.</dc:creator>
<dc:creator>Mineo, A.</dc:creator>
<dc:creator>Cortese, K.</dc:creator>
<dc:creator>Miguel-Aliaga, I.</dc:creator>
<dc:creator>Vignoli, A.</dc:creator>
<dc:creator>Borghi, E.</dc:creator>
<dc:creator>Galeone, A.</dc:creator>
<dc:creator>vaccari, t.</dc:creator>
<dc:date>2025-12-18</dc:date>
<dc:identifier>doi:10.64898/2025.12.16.694631</dc:identifier>
<dc:title><![CDATA[Mesodermal-specific MECP2 expression in Drosophila induces visceral and skeletal muscle defects rescued by butyrate supplementation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.12.17.694965v1?rss=1">
<title>
<![CDATA[
Capsid flexibility during Ty1 virus-like particle assembly 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.12.17.694965v1?rss=1"
</link>
<description><![CDATA[
The Ty1/Copia family (Pseudoviridae) is a widely studied group of long terminal repeat (LTR) retrotransposons present in diverse eukaryotes that share a common ancestor with retroviruses. The founding member Ty1 of Saccharomyces assembles into heterogeneous virus-like particles (VLPs) that are composed of the Gag and Gag-Pol proteins. We used cryo-electron tomography (cryo-ET) of Ty1 VLPs purified from a Ty1-less yeast strain expressing a functional Ty1 element to determine the organization and interactions of Gag in Ty1 VLPs. Using sub-tomogram averaging (STA), we observed unique capsomere structures and obtained EM density maps for pentagonal (7.5 [A]) and hexagonal (7.9 [A]) capsomeres. Through iterative rounds of alignment and classification, we demonstrate that Ty1 VLPs are composed of diverse arrangements of capsomeres, unlike true icosahedral virions. We modeled the structure of the conserved Ty1 Gag capsid C-terminal domain (CA-CTD), previously determined using X-ray crystallography into the capsomeric arrangements, and identified roles for the Dimer-1 interface present in the asymmetric unit, as well as additional interfaces involved in intra- and inter-capsomere assembly. We show that pentagonal or hexagonal capsomere assembly results from flexibility of CA-CTD - CA-CTD interactions at Interface-2. Additionally, we determined the structure of the Ty1 Gag capsid N-terminal domain (CA-NTD) using solution NMR and fitted the CA-NTD into the cryo-ET density maps. Our results indicate that the heterogeneous population of VLP size and morphology arises, at least in part, from the ability of the capsomeres to assemble in diverse organizations. (238 words)
]]></description>
<dc:creator>Sibert, B. S.</dc:creator>
<dc:creator>Hannon-Hatfield, A.</dc:creator>
<dc:creator>Nicastro, G.</dc:creator>
<dc:creator>Cottee, M. A.</dc:creator>
<dc:creator>Taylor, I. A.</dc:creator>
<dc:creator>Garfinkel, D. J.</dc:creator>
<dc:creator>Wright, E. R.</dc:creator>
<dc:date>2025-12-18</dc:date>
<dc:identifier>doi:10.64898/2025.12.17.694965</dc:identifier>
<dc:title><![CDATA[Capsid flexibility during Ty1 virus-like particle assembly]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.12.16.694583v1?rss=1">
<title>
<![CDATA[
BATF3 controls the homeostatic maintenance and function of innate-like γδ T cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.12.16.694583v1?rss=1"
</link>
<description><![CDATA[
{gamma}{delta} T cells compose an evolutionarily conserved lineage of lymphocytes, with both adaptive- and innate-like characteristics, contributing to tissue homeostasis, immune surveillance, and rapid responses to stress and infection. While their functional diversity and tissue-specific roles are tightly regulated by transcriptional networks, the underlying molecular mechanisms remain incompletely understood. The transcription factor basic leucine zipper ATF-like transcription factor 3 (BATF3) plays a central role in the development of conventional type 1 dendritic cells (cDC1s). Here, we unveil BATF3 as a critical cell-intrinsic regulator of the homeostasis, functional specialization, and tissue distribution of {gamma}{delta} T cells. Batf3-deficient mice display an altered composition of {gamma}{delta} T cell subsets, with a marked decrease in the numbers of innate-like {gamma}{delta} T cells across multiple organs when compared to their wild-type counterparts, independently of cDC1s. Loss of BATF3 impacts not only cell survival but also IL-17 production after {gamma}{delta} T cells complete their thymic development. Mechanistically, Batf3-deficient innate-like {gamma}{delta} T cells exhibit transcriptional changes that disrupt pathways governing actin cytoskeleton remodelling, immunological synapse organization and cellular identity. Notably, Batf3-deficient mice present decreased survival in a viral infection model highly dependent on innate-like {gamma}{delta} T cells. Together, our findings uncover a previously unrecognized BATF3-dependent pathway that controls {gamma}{delta} T cell morphology and function, profoundly impacting their biology.
]]></description>
<dc:creator>Papotto, P. H.</dc:creator>
<dc:creator>Hernandez-Garcia, E.</dc:creator>
<dc:creator>Zarco-Cuadrillero, A.</dc:creator>
<dc:creator>Redondo-Urzainqui, A.</dc:creator>
<dc:creator>Parkinson, J. E.</dc:creator>
<dc:creator>Pinheiro, R. G. R.</dc:creator>
<dc:creator>MacDonald, A.</dc:creator>
<dc:creator>Sancho, D.</dc:creator>
<dc:creator>Silva-Santos, B.</dc:creator>
<dc:creator>Allen, J. E.</dc:creator>
<dc:creator>Hayday, A. C.</dc:creator>
<dc:creator>Iborra, S.</dc:creator>
<dc:creator>Munoz-Ruiz, M.</dc:creator>
<dc:date>2025-12-18</dc:date>
<dc:identifier>doi:10.64898/2025.12.16.694583</dc:identifier>
<dc:title><![CDATA[BATF3 controls the homeostatic maintenance and function of innate-like γδ T cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.12.15.694339v1?rss=1">
<title>
<![CDATA[
PKN2 regulates cell-junctions to limit colitis and colon tumour formation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.12.15.694339v1?rss=1"
</link>
<description><![CDATA[
Background and AimsEpithelial barrier integrity in the gut must be tightly regulated to allow essential paracellular transport while limiting mucosal exposure to inflammatory microbes and toxins. Chronic inflammation in inflammatory bowel diseases is a major health burden and a key risk factor for colorectal cancer. Here, we identify the Rho-regulated kinase PKN2 as a non-redundant safeguard of epithelial barrier integrity and a candidate tumour suppressor in colorectal cancer.

MethodsConditional PKN2 knockout mouse models were used to examine the impact of PKN2 loss on colitis severity and induction of colorectal adenomas. Intestinal organoid models, epithelial barrier assays and bioinformatic approaches explored PKN2 regulation of epithelial function. Human inflammatory bowel disease datasets are analysed for associations with PKN2 expression and signatures associated with compromised barrier integrity.

ResultsPKN2 deletion sensitises mice to inflammatory bowel injury and promotes colorectal adenoma formation in a colitis-associated colorectal cancer model. Tumour burden tightly correlates with colitis severity suggesting that enhanced inflammation-driven tissue injury underlies tumour promotion. Mechanistically, PKN2 localises to epithelial tight junctions and is required to stabilise barrier integrity during epithelial injury in mouse and organoid models. Transcriptomic changes associated with epithelial PKN2 loss mimic inflammatory bowel diseases and correlate with diseases severity and therapy response in human datasets.

ConclusionsPKN2 is a regulator of gut barrier integrity encoded at genomic loci previously associated with sensitivity to bowel inflammation and tumour suppression in both humans and mice. PKN2 loss recapitulates key features of human colitis and implicates PKN2 as a regulator tight junction integrity that shapes disease severity, treatment response and cancer risk.
]]></description>
<dc:creator>Henry, J. C.</dc:creator>
<dc:creator>Campbell, A. N.</dc:creator>
<dc:creator>Huddar, P.</dc:creator>
<dc:creator>Sewell, J.</dc:creator>
<dc:creator>Menezes, S.</dc:creator>
<dc:creator>Passman, A.</dc:creator>
<dc:creator>Smith, K.</dc:creator>
<dc:creator>McGauran, S.</dc:creator>
<dc:creator>Lin, M.-L.</dc:creator>
<dc:creator>Rosewell, I.</dc:creator>
<dc:creator>Jansen, M.</dc:creator>
<dc:creator>McDonald, N. Q.</dc:creator>
<dc:creator>Graham, T. A.</dc:creator>
<dc:creator>McDonald, S. A.</dc:creator>
<dc:creator>Cameron, A. J.</dc:creator>
<dc:date>2025-12-17</dc:date>
<dc:identifier>doi:10.64898/2025.12.15.694339</dc:identifier>
<dc:title><![CDATA[PKN2 regulates cell-junctions to limit colitis and colon tumour formation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.01.05.697593v1?rss=1">
<title>
<![CDATA[
Humanized Kcnv2 E151X Mouse Captures Hallmarks of KCNV2-Associated Retinal Dystrophy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.01.05.697593v1?rss=1"
</link>
<description><![CDATA[
BackgroundKCNV2-associated retinopathy is a rare inherited retinal dystrophy caused by variants in the KCNV2 gene, leading to disrupted photoreceptor function and slowly progressive vision loss. Patients have characteristic electroretinography abnormalities, including reduced cone responses, delayed and reduced rod responses to low light flashes and paradoxally large rod-driven responses to bright flashes of light. To model this condition, we generated the Kcnv2 E151X mouse line and assessed its structural and functional retinal features.

MethodsWe have employed CRISPR/Cas 9 gene editing technology to generate a mouse line with an early stop mutation in position E151- orthologous to the commonly encountered E143X mutation in humans - and performed a combination of Iimmunohistochemistry and Western Blot to confirm the absence of the full-length KCNV2-encoded protein, Kv8.2. To assess how closely it models the human disease, we have characterised the KCNV2 mutant mouse line at histological and functional levels employing immunohistochemistry and electroretinography, respectiveley.

ResultsKcnv2 mutant mice showed markedly reduced photopic responses and reproduced the supernormal rod phenotype as described in affected individuals. In the morphological context, mutant retinas demonstrated strong and early glial fibrillary acidic protein upregulation together with reduced counts of cone arrestin positive cells as well as photoreceptors in general. Power calculations based on the data obtained herein suggest therapeutic trials are feasible with small sample sizes.

ConclusionsThe Kcnv2 mutant mouse line replicates key functional and structural hallmarks of KCNV2- associated retinopathy. This model provides a relevant platform for mechanistic studies and preclinical evaluation of gene based or pharmacological therapies targeting cone and rod photoreceptor dysfunction.
]]></description>
<dc:creator>Xhaferri, N.</dc:creator>
<dc:creator>Biswas, S.</dc:creator>
<dc:creator>Davies, B.</dc:creator>
<dc:creator>Lindner, M.</dc:creator>
<dc:date>2026-01-05</dc:date>
<dc:identifier>doi:10.64898/2026.01.05.697593</dc:identifier>
<dc:title><![CDATA[Humanized Kcnv2 E151X Mouse Captures Hallmarks of KCNV2-Associated Retinal Dystrophy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.01.02.697435v1?rss=1">
<title>
<![CDATA[
Dlx2 reprograms the transcriptome and laminar position of glia-derivedAscl1-induced interneurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.01.02.697435v1?rss=1"
</link>
<description><![CDATA[
Direct lineage reprogramming of glial cells into neurons offers a promising strategy to repair diseased brain circuits, but engineering defined neuronal subtypes remains challenging. We found that a phospho-site-deficient Ascl1 variant, Ascl1SA6, but not wildtype Ascl1, induces hallmarks of parvalbumin fast-spiking interneurons, raising the question of how closely these induced neurons resemble canonical cortical interneurons and what transcriptional events underlie this process. Single-cell transcriptomic analysis revealed that Ascl1SA6-induced neurons only partially recapitulated canonical interneuron programs and failed to induce the transcription factor Dlx2 and its downstream targets. Co-expression of Dlx2 with Ascl1SA6 restored a more canonical interneuron-like transcriptome, including genes involved in migration, and resulted in neurons occupying laminar positions more typical of endogenous interneurons. These findings provide molecular insights into how Ascl1 posttranslational modifications regulate its transcriptional activity and demonstrate a strategy to engineer induced cortical interneurons that more closely resemble their native counterparts, offering a framework for layer-specific restoration of inhibitory circuits in neurological diseases.
]]></description>
<dc:creator>Cooper, A.</dc:creator>
<dc:creator>Garcia Mora, A.</dc:creator>
<dc:creator>Herrera-Oropeza, G.</dc:creator>
<dc:creator>Beltran Arranz, A.</dc:creator>
<dc:creator>Marichal, N.</dc:creator>
<dc:creator>Shi, Y.</dc:creator>
<dc:creator>Leaman, S.</dc:creator>
<dc:creator>Lozano Gonzalez, K.</dc:creator>
<dc:creator>Strom, M.</dc:creator>
<dc:creator>Fursham, H.</dc:creator>
<dc:creator>Guillemot, F.</dc:creator>
<dc:creator>Berninger, B.</dc:creator>
<dc:date>2026-01-02</dc:date>
<dc:identifier>doi:10.64898/2026.01.02.697435</dc:identifier>
<dc:title><![CDATA[Dlx2 reprograms the transcriptome and laminar position of glia-derivedAscl1-induced interneurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.12.15.694413v1?rss=1">
<title>
<![CDATA[
Control of cellular cortical tension and shape by RhoGTPase signalling 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.12.15.694413v1?rss=1"
</link>
<description><![CDATA[
Shape changes are ubiquitous in biology, from cytokinesis at the single cell scale to tissue-scale morphogenesis involving coordinated changes in hundreds of cells. In all cases, morphogenesis is powered by gradients in mechanical tension that arise downstream of signalling. Many pathways converge on RhoGTPases that modulate the cytoskeleton and cell contractility to control cell mechanics and, subsequently, shape. Despite their physiological importance, we lack a quantitative understanding of how changes in signalling alter cortical mechanics to drive cell shape change. Here, we use optogenetics to quantitatively characterise the relationship between the amount of RhoGEF localised to the membrane, the downstream myosin recruitment, and the subsequent mechanical changes. We then show that cortical myosin amount and cortical tension increase linearly with the amount of membranous RhoGEF signalling. Based on these data, we develop a predictive model of the temporal evolution of RhoGEF membrane localisation, cortical myosin enrichment, and cortical tension in response to a pulse of light. Using this model together with an active surface model of the cell cortex, we show that the cellular shape changes induced by localised optogenetic recruitment of RhoGEF signalling can be predicted, directly linking gradients in signalling to shape change.

Significance statementShape changes are ubiquitous in biology, during division in single cells and in tissue during embryogenesis. In all cases, shape change is powered by gradients in mechanical tension that arise downstream of changes in biochemical signals. Despite their importance, we lack a quantitative understanding of how changes in signals alter cell mechanics to drive cell shape change. Here, we control the location and amount of biochemical signal using light to quantitatively characterise the relationship between signals and their resulting biological and mechanical changes. We show that mechanical change scales linearly with the amount of biochemical signal. Based on this, we develop a mathematical model that predicts cell mechanical and shape changes from the location and amount of biochemical signals.
]]></description>
<dc:creator>Bohec, P.</dc:creator>
<dc:creator>Khoromskaia, D.</dc:creator>
<dc:creator>Kelkar, M.</dc:creator>
<dc:creator>Ferber, E.</dc:creator>
<dc:creator>Duprez, G.</dc:creator>
<dc:creator>Lavoie, G.</dc:creator>
<dc:creator>Valon, L.</dc:creator>
<dc:creator>Roux, P. P.</dc:creator>
<dc:creator>Salbreux, G.</dc:creator>
<dc:creator>Charras, G.</dc:creator>
<dc:date>2025-12-17</dc:date>
<dc:identifier>doi:10.64898/2025.12.15.694413</dc:identifier>
<dc:title><![CDATA[Control of cellular cortical tension and shape by RhoGTPase signalling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.01.12.699057v1?rss=1">
<title>
<![CDATA[
The Structured RNA-binding Domains and Condensation Capacity of FUS Shape its RNA-binding Landscape and Function. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.01.12.699057v1?rss=1"
</link>
<description><![CDATA[
RNA-binding proteins (RBPs) are key regulators of gene expression and often contain intrinsically disordered regions that drive biomolecular condensation. Yet, how condensation affects RBP specificity and function remains poorly defined. Here, we use strategically designed point mutations to selectively impair canonical RNA-binding and condensation of the amyotrophic lateral sclerosis (ALS)-linked RBP Fused in Sarcoma (FUS). Using automated high-content imaging, we show that both properties shape nuclear ribonucleoprotein condensates and govern distinct aspects of FUS function in the DNA damage response. Transcriptome-wide mapping of FUS-RNA interactions reveals that the canonical RNA-binding domains recognise G-rich and C-rich motifs, whereas condensation selectively enhances binding to G-rich and structured sequences, often acting in concert with canonical RNA-binding to control transcriptional programs and splicing decisions. These findings provide a mechanistic framework for how FUS integrates condensation with sequence-specific RNA recognition to orchestrate nuclear organisation, genome stability and RNA metabolism. Furthermore, our rigorously validated mutants offer a platform for future mechanistic dissection of ALS pathogenesis and the development of targeted therapeutic strategies.

HighlightsO_LIFUS condensation and RNA-binding can be functionally uncoupled with targeted point mutations
C_LIO_LICondensation drives FUS recruitment and assembly of nuclear RNP condensates
C_LIO_LIFUS condensation and RNA-binding play distinct roles in the DNA damage response
C_LIO_LIThe structured RNA-binding domains confer specificity to G-rich and C-rich motifs
C_LIO_LICondensation enhances FUS binding to G-rich, structured RNA elements
C_LIO_LIFUS condensation and canonical RNA-binding synergistically regulate gene expression
C_LI
]]></description>
<dc:creator>Jutzi, D.</dc:creator>
<dc:creator>Alcalde, J.</dc:creator>
<dc:creator>Hutten, S.</dc:creator>
<dc:creator>Tiryaki, F.</dc:creator>
<dc:creator>Davies, B.</dc:creator>
<dc:creator>Plun-Favreau, H.</dc:creator>
<dc:creator>Sibley, C.</dc:creator>
<dc:creator>Dormann, D.</dc:creator>
<dc:creator>Ruepp, M.-D.</dc:creator>
<dc:date>2026-01-12</dc:date>
<dc:identifier>doi:10.64898/2026.01.12.699057</dc:identifier>
<dc:title><![CDATA[The Structured RNA-binding Domains and Condensation Capacity of FUS Shape its RNA-binding Landscape and Function.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.01.12.698986v1?rss=1">
<title>
<![CDATA[
Bro1-Mediated Trafficking Couples TOR Signalling to Cellular Metabolism and Longevity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.01.12.698986v1?rss=1"
</link>
<description><![CDATA[
Adaptation to nutrient availability requires coordination between growth control, metabolism, and intracellular trafficking. In eukaryotes, inhibition of Target of Rapamycin (TOR) signalling robustly promotes stress resistance and longevity, yet how reduced growth signalling is coupled to organelle dynamics and proteome remodelling remains unclear. Here, we identify the conserved ESCRT-associated protein Bro1 as a central integrator of TOR signalling, vacuolar trafficking, and metabolic adaptation. Using fission yeast, we show that Bro1 is required for normal lifespan and for the global proteomic reprogramming that accompanies TOR inhibition. In Bro1 mutant cells, repression of ribosome biogenesis is uncoupled from activation of catabolic, vacuolar, and metabolic pathways, resulting in an altered metabolic state characterised by elevated lipid metabolism and increased abundance of nutrient transporters.

Mechanistically, Bro1 promotes TOR-dependent cargo deubiquitination, vacuolar trafficking, and turnover of plasma membrane hexose transporters and enables appropriate nuclear relocalisation of the transcriptional repressor Scr1. In the absence of Bro1, nutrient transporters persist at the cell surface despite TOR inhibition, conferring resistance to TOR inhibitors while impairing stress responses and reducing lifespan. Together, our findings establish Bro1 as a key coordinator linking ESCRT-mediated endosomal-vacuolar trafficking to TOR-dependent metabolic control. By coupling growth suppression to enhanced recycling and cellular maintenance, Bro1 enables the transition from growth to longevity-promoting states, revealing a mechanism connecting intracellular trafficking, metabolism, and ageing.
]]></description>
<dc:creator>Ng, K.</dc:creator>
<dc:creator>Kumar, J.</dc:creator>
<dc:creator>Dabrowska, A.</dc:creator>
<dc:creator>Clemente-Ramos, J.</dc:creator>
<dc:creator>Islam, R. A.</dc:creator>
<dc:creator>Xintarakou, O.</dc:creator>
<dc:creator>Freiwald, A.</dc:creator>
<dc:creator>Barthel, N.</dc:creator>
<dc:creator>Ludwig, D.</dc:creator>
<dc:creator>Stamataki, D.</dc:creator>
<dc:creator>Alao, J. P.</dc:creator>
<dc:creator>Thorpe, P.</dc:creator>
<dc:creator>Ralser, M.</dc:creator>
<dc:creator>Mulleder, M.</dc:creator>
<dc:creator>Mole, S. E.</dc:creator>
<dc:creator>Rallis, C.</dc:creator>
<dc:date>2026-01-12</dc:date>
<dc:identifier>doi:10.64898/2026.01.12.698986</dc:identifier>
<dc:title><![CDATA[Bro1-Mediated Trafficking Couples TOR Signalling to Cellular Metabolism and Longevity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.01.09.698651v1?rss=1">
<title>
<![CDATA[
Genetic sex of enteric neurons enables ovarian relaxin togate maternal gut plasticity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.01.09.698651v1?rss=1"
</link>
<description><![CDATA[
Animals must align intestinal plasticity and feeding with reproductive state, yet the checkpoint that gates these adaptations is unknown. Here we show that an ovary-to-enteric-neuron axis gates the onset of maternal gut plasticity in Drosophila. Genetic sex establishes endocrine competence in a subset of enteric neurons via the sex determination pathway, enabling female-specific expression of the relaxin-family receptor Lgr3. After mating, steroid signalling increases Lgr3 receptor expression, priming these neurons for reproductive adaptation. Once oocytes mature fully, follicle cells secrete the relaxin-like hormone dILP8, which activates Lgr3 to trigger gut enlargement and increased feeding. Disrupting the sex determination pathway in enteric neurons, Lgr3, or ovarian dILP8 prevents gut enlargement and reduces feeding. Thus, genetic sex establishes competence, steroid signalling primes it, and ovarian relaxin triggers it, defining a maternal intestinal plasticity checkpoint that ensures gut adaptations initiate only once reproduction is underway and energy demands peak. Our findings delineate an ovary-to-enteric-neuron axis that couples reproductive state to maternal gut plasticity.
]]></description>
<dc:creator>Silva, B.</dc:creator>
<dc:creator>Tsakani, E.</dc:creator>
<dc:creator>De, J.</dc:creator>
<dc:creator>Kontopoulos, A.</dc:creator>
<dc:creator>Gaspar, P.</dc:creator>
<dc:creator>Hiridjee, A.</dc:creator>
<dc:creator>Rosselli, C.</dc:creator>
<dc:creator>de Juan-Sanz, J.</dc:creator>
<dc:creator>Campusano, J. M.</dc:creator>
<dc:creator>Treiber, C. D.</dc:creator>
<dc:creator>Waddell, S.</dc:creator>
<dc:creator>Miguel-Aliaga, I.</dc:creator>
<dc:creator>Hadjieconomou, D.</dc:creator>
<dc:date>2026-01-10</dc:date>
<dc:identifier>doi:10.64898/2026.01.09.698651</dc:identifier>
<dc:title><![CDATA[Genetic sex of enteric neurons enables ovarian relaxin togate maternal gut plasticity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.01.16.699845v1?rss=1">
<title>
<![CDATA[
Iterative Bump-and-hole engineering creates a bioorthogonal reporter for N-acetylglucosaminyltransferase I 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.01.16.699845v1?rss=1"
</link>
<description><![CDATA[
Asparagine-linked protein glycosylation is among the most frequent modifications of proteins trafficking through the secretory pathway. These glycans are manufactured in an assembly line process to a common precursor that is then subject to individual modifications with different levels of complexity. An important biosynthetic modulator is the incorporation of N-acetylglucosamine (GlcNAc) at distinct positions in N-linked glycan biosynthesis, commencing with the activity of the glycosyltransferase MGAT1. While mapping of N-glycans to their corresponding protein attachment sites is generally possible, not much is known about the glycoprotein substrate choice for MGAT1 and related transferases. Analogs of GlcNAc with small bioorthogonal tags can be incorporated into N-glycans. However, due to the promiscuity of some GlcNAc transferases, incorporation is of little specificity towards individual positions. Here, we report an iterative bump-and-hole approach in the design of a bioorthogonal precision tool for the activity of MGAT1 in mammalian cells. Structure-informed protein engineering abrogated the activity of MGAT1 towards the nucleotide-sugar UDP-GlcNAc while retaining activity towards bumped, azide-modified analogs. Kinetic and computational analyses using a neural network approach informed the synthesis of a tailored UDP-GlcNAc analog with preferential acceptance by the engineered enzyme. Following substrate biosynthesis, the strategy allowed selective incorporation of a chemical tag on MGAT1 substrate proteins in living mammalian cells with little background incorporation by other GlcNAc transferases. Our work expands the toolbox for glycan-based reporter compounds.
]]></description>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Pieters, S.</dc:creator>
<dc:creator>Bineva-Todd, G.</dc:creator>
<dc:creator>Sagiroglugil, M.</dc:creator>
<dc:creator>Burnap, S. A.</dc:creator>
<dc:creator>Hoddle, F.</dc:creator>
<dc:creator>Cioce, A.</dc:creator>
<dc:creator>Ohara, A.</dc:creator>
<dc:creator>Bruemmer, K.</dc:creator>
<dc:creator>Bertozzi, C.</dc:creator>
<dc:creator>Polizzi, K. M.</dc:creator>
<dc:creator>Struwe, W. B.</dc:creator>
<dc:creator>Rovira, C.</dc:creator>
<dc:creator>Schumann, B.</dc:creator>
<dc:date>2026-01-17</dc:date>
<dc:identifier>doi:10.64898/2026.01.16.699845</dc:identifier>
<dc:title><![CDATA[Iterative Bump-and-hole engineering creates a bioorthogonal reporter for N-acetylglucosaminyltransferase I]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.01.19.700281v1?rss=1">
<title>
<![CDATA[
SARS-CoV-2 membrane protein biogenesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.01.19.700281v1?rss=1"
</link>
<description><![CDATA[
Viral protein biogenesis underpins every viral life cycle stage, and elucidating these processes could reveal fundamental principles of virus-host interaction, and vulnerabilities amenable to therapeutic targeting. Here we apply biophysical, molecular, and cell biology techniques to investigate the insertion, folding, and oligomerization of the SARS-CoV-2 M protein. We describe the sequential co-translational insertion of the hydrophobic core, and demonstrate that the cytosolic C-terminal domain undergoes slower adoption of its tertiary structure. Additionally, we characterize how the transmembrane domain bundle facilitates M-protein oligomerization. Our results reveal a hydrophobic residue cluster that is essential for protein folding and co-translational dimerization. Additionally, we identify the cellular machinery responsible for targeting and inserting the M protein into the ER membrane, and chaperones and cofactors that may contribute to proper folding.
]]></description>
<dc:creator>Ortiz Mateu, J.</dc:creator>
<dc:creator>Pearson, G. J.</dc:creator>
<dc:creator>Rius-Salvador, M.</dc:creator>
<dc:creator>Sedighian, S.</dc:creator>
<dc:creator>Pavlova, A.</dc:creator>
<dc:creator>Alonso-Romero, J.</dc:creator>
<dc:creator>Acosta-Caceres, J. M.</dc:creator>
<dc:creator>Metola, A.</dc:creator>
<dc:creator>Garcia-Murria, M. J.</dc:creator>
<dc:creator>Skehel, J. M.</dc:creator>
<dc:creator>Gumbart, J. C.</dc:creator>
<dc:creator>Carlton, J. C.</dc:creator>
<dc:creator>von Heijne, G.</dc:creator>
<dc:creator>Sanchez-del Pino, M. M.</dc:creator>
<dc:creator>Martinez-Gil, L.</dc:creator>
<dc:creator>Mingarro, I.</dc:creator>
<dc:date>2026-01-20</dc:date>
<dc:identifier>doi:10.64898/2026.01.19.700281</dc:identifier>
<dc:title><![CDATA[SARS-CoV-2 membrane protein biogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.01.23.697869v1?rss=1">
<title>
<![CDATA[
Essential function reflected in the phylodynamics of a multigene family: the pir genes of malaria parasites 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.01.23.697869v1?rss=1"
</link>
<description><![CDATA[
The genomes of malaria parasites (Plasmodium spp.) encode many gene families, which are intimately associated with host interactions and disease in these important pathogens. The largest malaria gene family is the Plasmodium interspersed repeat (pir) genes, present in rodent, primate and most human malaria parasites, which are suggested to have originated from one highly conserved gene, which we call pirC1. The precise function(s) of pir is unknown but to determine their potentially multifarious roles we must understand the evolutionary dynamics of pir repertoire to discriminate among the many genes. Here we estimate the global phylogeny for pir genes in 14 Plasmodium species and one Hepatocystis species. We reveal that pirC1 is not the common ancestor but is one of several orthologous genes conserved in multiple species amidst the rapid turnover of species-specific paralogs. We show that the PIRC1 protein is nonetheless essential for blood stage growth of P. berghei, P. chabaudi and P. knowlesi, as parasites lacking the pirC1 gene could not be generated or had severely reduced growth rates. As this effect was observed both in vivo and in vitro, the role of pirC1 is not related to host immune interaction. Rather, P. berghei and P. knowlesi PIRC1 are secreted from the parasite, pointing to a role in parasite interaction with the host cell or nutrient uptake by blood stages. The phylodynamics of pir genes indicate that old orthologs, like pirC1, and younger within-species paralogs could have fundamentally different roles, and emphasize the need to distinguish between them in future. This study is the first to provide evidence for the existence of an essential pir gene and provides a robust rationale for further experimental approaches to pir gene functions.

SIGNFICANCEThe genomes of malaria parasites (Plasmodium) contain many different gene families, of which the pir family is the largest, with more than 1000 members in some species. The PIR proteins are likely important for parasite fitness but their precise functions remain unknown - roles in adherence of infected red blood cells to blood vessels, virulence and immune evasion of have been suggested. How, and why, this highly diverse gene family evolved is a significant question both for understanding malaria physiology and pathogenesis. Here we present a comprehensive pir phylogeny, identifying the origins of gene diversity during Plasmodium evolution and a select group of highly conserved genes. We show that one conserved pir gene (pirC1) encodes a protein that is essential for optimal growth of multiple malaria parasites during the asexual blood stage, both in the host and in vitro. This indicates that pirC1 function relates to interaction with the host cell or nutrient acquisition, and not to immune evasion or sequestration, (although this might still be the function of other pir genes). This study provides a robust rationale for the hitherto baffling diversity of pir genes, and shows why it is important to distinguish old orthologs from young paralogs in future studies on pir gene function.
]]></description>
<dc:creator>Jackson, A. P.</dc:creator>
<dc:creator>Cunningham, D. A.</dc:creator>
<dc:creator>Lin, L.</dc:creator>
<dc:creator>Claro de Oliveira, N. M.</dc:creator>
<dc:creator>Chevalley-Maurel, S. C.</dc:creator>
<dc:creator>Pianta, G.</dc:creator>
<dc:creator>Little, T. S.</dc:creator>
<dc:creator>Mohring, F.</dc:creator>
<dc:creator>Renfree, A. K.</dc:creator>
<dc:creator>Moon, R. W.</dc:creator>
<dc:creator>Langhorne, J. W.</dc:creator>
<dc:creator>Janse, C. J.</dc:creator>
<dc:creator>Franke-Fayard, B. M. D.</dc:creator>
<dc:creator>van Ooij, C.</dc:creator>
<dc:date>2026-01-23</dc:date>
<dc:identifier>doi:10.64898/2026.01.23.697869</dc:identifier>
<dc:title><![CDATA[Essential function reflected in the phylodynamics of a multigene family: the pir genes of malaria parasites]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.01.29.702321v1?rss=1">
<title>
<![CDATA[
Ex vivo maturation of the malaria parasite egress protease SERA6 aids pathway dissection and inhibitor development. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.01.29.702321v1?rss=1"
</link>
<description><![CDATA[
Release (egress) of malaria parasites from host red blood cells (RBC) is a protease-dependent process involving breakdown of the RBC cytoskeleton by a parasite cysteine protease-like protein called SERA6. In the penultimate step of the egress cascade, SERA6 undergoes autoproteolytic maturation triggered upon cleavage by a serine protease called SUB1 and requiring interactions between SERA6 and fragments of another parasite protein called MSA180. Egress can be blocked by treatment of intraerythrocytic parasites with small molecules that prevent the autocatalytic SERA6 maturation step, suggesting that SERA6 is a druggable target. Here we describe the development of a cell-free in vitro system that recapitulates SERA6 maturation. We use the assay to confirm the strict requirement for MSA180 in SERA6 maturation by SUB1 and to show that these 3 components are sufficient for SERA6 maturation. Using a synthetic peptide substrate based on a predicted autocatalytic cleavage site we demonstrate that the fully mature SERA6 is an active proteolytic enzyme and we validate improved small molecule inhibitors of SERA6. Our lead inhibitory compound efficiently blocks egress of asexual blood stage parasites, confirming SERA6 as a new potential antimalarial drug target.
]]></description>
<dc:creator>Withers-Martinez, C.</dc:creator>
<dc:creator>Taha, Z.</dc:creator>
<dc:creator>Collins, C. R.</dc:creator>
<dc:creator>Hackett, F.</dc:creator>
<dc:creator>Tan, M. S.</dc:creator>
<dc:creator>Soudy, C.</dc:creator>
<dc:creator>Joshi, D.</dc:creator>
<dc:creator>Redmond, J.</dc:creator>
<dc:creator>Davies, B.</dc:creator>
<dc:creator>Maslen, S.</dc:creator>
<dc:creator>Skehel, M.</dc:creator>
<dc:creator>George, R.</dc:creator>
<dc:creator>Kjaer, S.</dc:creator>
<dc:creator>Blackman, M. J.</dc:creator>
<dc:date>2026-01-29</dc:date>
<dc:identifier>doi:10.64898/2026.01.29.702321</dc:identifier>
<dc:title><![CDATA[Ex vivo maturation of the malaria parasite egress protease SERA6 aids pathway dissection and inhibitor development.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.01.26.701684v1?rss=1">
<title>
<![CDATA[
ESFS: A Noise-Resilient Framework for Feature Selection and Marker Gene Discovery in Single-Cell Transcriptomics 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.01.26.701684v1?rss=1"
</link>
<description><![CDATA[
Single-cell RNA sequencing (scRNA-seq) has transformed our ability to resolve cellular heterogeneity, but extracting meaningful signals remains challenging due to technical noise, batch effects, and the limitations of current feature selection methods. We present Entropy Sorting Feature Selection (ESFS), a modular, user-friendly framework that captures multivariate gene expression relationships without imputation or denoising via latent spaces. Across diverse datasets, ESFS improves interpretability and reveals biology missed by standard workflows: identifying coherent developmental programs in eight independent human embryo datasets without batch integration; resolving spatial gene expression in mouse colon obscured by conventional analyses; distinguishing shared and tumour-specific microenvironments in glioblastoma; and disambiguating spatial, temporal, and neurogenic programs in the developing mouse neural tube. By operating in gene expression space, ESFS produces interpretable, biologically meaningful outputs while reducing artefacts introduced by feature extraction. These results position ESFS as a powerful means to uncover relevant molecular signatures in noisy, high-dimensional transcriptomics data.
]]></description>
<dc:creator>Radley, A.</dc:creator>
<dc:creator>Boezio, G.</dc:creator>
<dc:creator>Shand, C.</dc:creator>
<dc:creator>Perez-Carrasco, R.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:date>2026-01-27</dc:date>
<dc:identifier>doi:10.64898/2026.01.26.701684</dc:identifier>
<dc:title><![CDATA[ESFS: A Noise-Resilient Framework for Feature Selection and Marker Gene Discovery in Single-Cell Transcriptomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.24.678316v1?rss=1">
<title>
<![CDATA[
Mapping the molecular landscape of the living brain: integrating spatial transcriptomics and qMRI 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.24.678316v1?rss=1"
</link>
<description><![CDATA[
Profiling dynamic molecular processes in neurodegeneration in vivo remains a major clinical challenge. Limited brain tissue accessibility especially limits the development and effective deployment of emerging disease modifying therapies, highlighting the need for non-invasive methods for profiling molecular disease processes. We introduce a non-invasive imaging framework that integrates ultra-high-resolution 7T quantitative MRI (qMRI) with spatially-resolved transcriptomics (SRT) to infer cell-type and pathway-specific molecular features within the cortical grey matter. This proof-of-concept establishes the integration of qMRI and SRT as a scalable, non-invasive platform for molecular profiling in neurodegeneration, with significant potential for precision therapeutic monitoring, drug development and clinical trials.
]]></description>
<dc:creator>Grant-Peters, M.</dc:creator>
<dc:creator>Thomas, G. E. C.</dc:creator>
<dc:creator>Zarkali, A.</dc:creator>
<dc:creator>Dobreva, I.</dc:creator>
<dc:creator>Weil, R.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:date>2025-09-25</dc:date>
<dc:identifier>doi:10.1101/2025.09.24.678316</dc:identifier>
<dc:title><![CDATA[Mapping the molecular landscape of the living brain: integrating spatial transcriptomics and qMRI]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.01.29.702551v1?rss=1">
<title>
<![CDATA[
Immune signatures of protection and progression in the airways of recent human tuberculosis household contacts 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.01.29.702551v1?rss=1"
</link>
<description><![CDATA[
Whilst the majority of individuals infected with M. tuberculosis control the infection and remain asymptomatic, only 5-10% progress to active tuberculosis (TB)1. Previous or current infection with M. tuberculosis is detected using antigen-specific interferon (IFN)-{gamma} release assays (IGRA), which cannot identify those who will remain healthy or progress to active TB1. However, 18F-Fluorodeoxyglucose positron emission-computed tomography (PET-CT) can detect increased immune cell metabolic activity in lung parenchyma and intrathoracic lymph nodes associated with infection2. The local early immune factors dictating protection or disease progression have not been defined. To address this, we interrogated the airway immune response at single cell resolution in bronchoalveolar lavage (BAL) from extensively clinically characterised recent household contacts of TB patients, who either controlled the infection or progressed to TB disease. Using unbiased analysis of bulk and scRNA-seq of BAL samples, we define type I IFN-dependent and -independent neutrophil signatures in active TB patients and contacts that progressed to TB. We additionally report an inverse relationship between airway neutrophils and T cells, with T cells showing signatures of exhaustion, cytotoxicity and cell death in progressors and TB patients with a neutrophil dominated airway profile. Conversely, T cell signatures of protection in contacts who remained healthy were dominated by genes related to regulation, quiescence and a stem-like profile. We show that both the inflammatory neutrophil signature of TB progression and the stem-like T cell signature of non-progressors from human airways were recapitulated in scRNA-seq data from non-human primate (NHP) granulomas, associated with disease or immune protection, respectively. Our findings from early human airway responses in TB contacts reveal genes, pathways and cell states that may dictate infection outcome and inform strategies for host-directed therapy and vaccine studies.
]]></description>
<dc:creator>Branchett, W. J.</dc:creator>
<dc:creator>Kim, J.-W.</dc:creator>
<dc:creator>Shields, J.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Novsarka, I.</dc:creator>
<dc:creator>Slawinski, H.</dc:creator>
<dc:creator>Wilkinson, K. A.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:creator>Kamil, A.</dc:creator>
<dc:creator>Verma, R.</dc:creator>
<dc:creator>Haldar, P.</dc:creator>
<dc:creator>O'Garra, A.</dc:creator>
<dc:date>2026-02-02</dc:date>
<dc:identifier>doi:10.64898/2026.01.29.702551</dc:identifier>
<dc:title><![CDATA[Immune signatures of protection and progression in the airways of recent human tuberculosis household contacts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.01.29.702696v1?rss=1">
<title>
<![CDATA[
FluNexus: a versatile web platform for antigenic prediction and visualization of influenza A viruses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.01.29.702696v1?rss=1"
</link>
<description><![CDATA[
Influenza A viruses continuously undergo antigenic evolution to escape host immunity induced by previous infections or vaccinations, consequently causing seasonal epidemics and occasional pandemics. Antigenic prediction and visualization of influenza A viruses are crucial for precise vaccine strain selection and robust pandemic preparedness. However, a user-friendly online platform for these capabilities remains notably absent, despite widespread demand. Here, we present FluNexus (https://flunexus.com), the first-of-its-kind, one-stop-shop web platform designed to facilitate the prediction and visualization of the antigenic change in emerging variants. FluNexus features a data preprocessing module for hemagglutinin subunit 1 (HA1) and hemagglutination inhibition (HI) data across three major public health threat subtypes (H1, H3 and H5). Meanwhile, FluNexus provides an interactive interface for online antigenic prediction and offers practical guidance for researchers. Most notably, FluNexus offers the visualization of influenza A virus antigenic evolution, providing intuitive insights into its antigenic dynamics. Specially, FluNexus proposes a novel manifold-based method for positioning antigens and antisera, generating accurate antigenic cartographies even with sparse HI data. By alleviating the programming burden on biologists, FluNexus supports more informed decision-making in vaccine strain selection and strengthens surveillance and pandemic preparedness.

HighlightsO_LIFluNexus features a data preprocessing module for HA1 and HI data spanning the H1, H3, and H5 subtypes.
C_LIO_LIFluNexus facilitates online antigenic prediction utilizing ten state-of-the-art antigenic prediction tools, and offers practical guidance based on a comparative evaluation of their performance.
C_LIO_LIFluNexus provides a visualization module for mapping antigenic evolution of influenza A viruses, incorporating a novel manifold-based method for antigenic cartography.
C_LI
]]></description>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Zhou, C.</dc:creator>
<dc:creator>Wu, H.</dc:creator>
<dc:creator>Xiao, K.</dc:creator>
<dc:creator>Hao, J.</dc:creator>
<dc:creator>Zhao, D.</dc:creator>
<dc:creator>Zhu, J.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Peng, J.</dc:creator>
<dc:creator>Gu, J.</dc:creator>
<dc:creator>Deng, G.</dc:creator>
<dc:creator>Cai, W.</dc:creator>
<dc:creator>Li, M.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Shang, X.</dc:creator>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Kong, H.</dc:creator>
<dc:date>2026-01-30</dc:date>
<dc:identifier>doi:10.64898/2026.01.29.702696</dc:identifier>
<dc:title><![CDATA[FluNexus: a versatile web platform for antigenic prediction and visualization of influenza A viruses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.02.06.704498v1?rss=1">
<title>
<![CDATA[
Targeting Myc activates a tissue-specific tumour resolution programme 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.02.06.704498v1?rss=1"
</link>
<description><![CDATA[
Neoplastic transformation parallels hallmark cellular programs of tissue regeneration and wound repair1,2. However, the mechanisms driving tumour regression upon oncogenic driver inhibition--and why it often fails--remain poorly understood3-7. Oncogenic KRas mutations and Myc deregulation, two archetypal cancer drivers, frequently occur and cooperate to promote aggressive lung adenocarcinoma (LUAD)8-12. To investigate the mechanistic consequences of targeting Myc in LUAD, we applied spatiotemporally controlled genetic and functional perturbations in a reversible KRas/Myc-driven mouse model, integrated with RNA sequencing and immune protein profiling of tumours and their microenvironment. Acute oncogenic Myc inactivation in epithelial tumour cells elicits a localised regenerative immune response crucially dependent on rapid, transient release of the alarmin cytokine interleukin-33 (IL-33) by alveolar type 2 tumour cells. As a sentinel signal for Myc loss, IL-33 signalling reverses tumour immunosuppression and neoangiogenesis, critically recruits eosinophils, and promotes neoplastic cell elimination, driving regression beyond mere growth arrest. Notably, brief systemic recombinant IL-33 administration to mice with KRas/Myc-driven LUAD induces robust eosinophil influx and near-complete tumour resolution. Together, these findings demonstrate that blocking Myc activates an innate, tissue-intrinsic immune programme rooted in resolution of wound repair and capable of driving regression when activated in a tumour. This opens the possibility of treating cancer not only by blocking mitogenic oncogenic drivers but also by pro-actively triggering pro-resolution pathways.
]]></description>
<dc:creator>Kortlever, R. M.</dc:creator>
<dc:creator>Campos, T.</dc:creator>
<dc:creator>Boeing, S.</dc:creator>
<dc:creator>Uzwyshyn-Jones, K.</dc:creator>
<dc:creator>Perfetto, A.</dc:creator>
<dc:creator>Evan, G. I.</dc:creator>
<dc:date>2026-02-09</dc:date>
<dc:identifier>doi:10.64898/2026.02.06.704498</dc:identifier>
<dc:title><![CDATA[Targeting Myc activates a tissue-specific tumour resolution programme]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.02.12.705497v1?rss=1">
<title>
<![CDATA[
Position Dependent Feedback Drives Scaling and Robustness of Morphogen Gradients 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.02.12.705497v1?rss=1"
</link>
<description><![CDATA[
Developmental patterning is remarkably robust to intrinsic and extrinsic variation. Morphogen gradients are a key mechanism driving patterning, and themselves often scale with the size of developing tissues and exhibit robustness to other perturbations. Recent data indicates that expander molecules, thought to drive morphogen scaling through expansion-repression (ER) feedback, have concentration profiles that are position dependent. This challenges the currently accepted ER mechanism that requires uniform expander concentrations and position independent feedback. To reconcile these observations, we introduce a new ER motif that supports morphogen scaling with both uniform and position dependent expander concentrations. We quantify scaling as a function of position, and demonstrate that the spatial profiles of scaling and robustness to perturbations in morphogen production are highly correlated. In contrast to uniform expander concentrations that can confer high levels of scaling and robustness at a single position, position dependent expander concentrations can enhance both scaling and robustness throughout the entire target tissue. We explore trade-offs associated with the dynamic range of the expander concentration, revealing that it can be varied to tune the locations where morphogen gradients confer scaling, robustness and precision simultaneously. These findings offer new insight into how developmental systems balance competing demands to achieve reproducible patterning despite biological variability.
]]></description>
<dc:creator>Mosby, L. S.</dc:creator>
<dc:creator>Hadjivasiliou, Z.</dc:creator>
<dc:date>2026-02-16</dc:date>
<dc:identifier>doi:10.64898/2026.02.12.705497</dc:identifier>
<dc:title><![CDATA[Position Dependent Feedback Drives Scaling and Robustness of Morphogen Gradients]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.02.10.704996v1?rss=1">
<title>
<![CDATA[
Emergence and antigenic characterisation of influenza A(H3N2) viruses with hemagglutinin substitutions N158K and K189R during the 2024/25 influenza season 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.02.10.704996v1?rss=1"
</link>
<description><![CDATA[
BackgroundSeasonal human influenza viruses can escape from antibody-mediated neutralization when amino acid changes occur in the hemagglutinin protein. Routine surveillance identified circulation of an A(H3N2) virus variant in the Netherlands with amino acid substitutions at hemagglutinin positions 158 and 189. These amino acid positions were previously responsible for antigenic change of influenza A(H3N2) viruses and potentially lead to escape of this variant from vaccine-mediated immunity.

AimTo characterize the emergence and antigenic properties of N158K and K189R double substitution virus variants.

MethodsWe analyzed the geographical and temporal dynamics of the double-substitution variant using a phylogeographic approach and used hemagglutination inhibition assays and antigenic cartography methods to map its antigenic properties.

ResultsA(H3N2) viruses carrying K189R were first detected in Guatemala in June 2024, before subsequently gaining the N158K substitution, which was intially detected in Colombia in November 2024, followed by detection in the Netherlands in December 2024. However, detections within Europe remained almost entirely confined to the Netherlands. The proportion of viruses carrying the N158K and K189R substitutions increased to 16% - 24% per collection week of sequenced Dutch viruses during the peak of the epidemic of the 2024-2025 respiratory season. Antigenic characterization of viruses with N158K and K189R substitutions indicated that these are antigenically distinct from the A(H3N2) components of 2025-2026 Northern Hemisphere vaccines, showing 8-192-fold reduction in hemagglutination inhibition titers with antisera against the vaccine strain compared to antisera against the homologous virus.

ConclusionsInfluenza A(H3N2) viruses with N158K and K189R escaped recognition by antibodies raised against the 2024-2025 and 2025/2026 Northern Hemipshere vaccine strains in hemagglutination inhibition assays. These variants circulated widely in the Netherlands during the 2024-2025 influenza season, raising concerns about reduced vaccine-mediated protection if such variants would spread more broadly during 2025-2026 Northern Hemipshere season.
]]></description>
<dc:creator>Koel, B.</dc:creator>
<dc:creator>Byrne, A.</dc:creator>
<dc:creator>Turner, S.</dc:creator>
<dc:creator>James, S.</dc:creator>
<dc:creator>Harvey, R.</dc:creator>
<dc:creator>Galiano, M.</dc:creator>
<dc:creator>Carr, C.</dc:creator>
<dc:creator>Lexmond, P.</dc:creator>
<dc:creator>Pronk, M.</dc:creator>
<dc:creator>Liang, R.</dc:creator>
<dc:creator>Boons, G.-J.</dc:creator>
<dc:creator>de Vries, R.</dc:creator>
<dc:creator>Eggink, D.</dc:creator>
<dc:creator>Lewis, N.</dc:creator>
<dc:creator>Smith, D.</dc:creator>
<dc:creator>Meijer, A.</dc:creator>
<dc:creator>Fouchier, R.</dc:creator>
<dc:date>2026-02-10</dc:date>
<dc:identifier>doi:10.64898/2026.02.10.704996</dc:identifier>
<dc:title><![CDATA[Emergence and antigenic characterisation of influenza A(H3N2) viruses with hemagglutinin substitutions N158K and K189R during the 2024/25 influenza season]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.02.18.706657v1?rss=1">
<title>
<![CDATA[
Discrete genetic effects of VHL and PBRM1 inactivation co-operate to disrupt epithelial homeostasis and promote ccRCC 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.02.18.706657v1?rss=1"
</link>
<description><![CDATA[
Inactivation of VHL is a truncal alteration in clear cell renal cell carcinoma, but additional events are required for oncogenesis, most commonly PBRM1 inactivation. To better understand this co-operation, we used an oncogenic cell-tagging strategy to analyze the earliest transcriptional and cellular consequences of Vhl and/or Pbrm1 inactivation in the renal tubular epithelium, in vivo, at single-cell resolution. Pbrm1 inactivation did not globally alter HIF-dependent transcription or increase early tubular proliferation induced by Vhl inactivation. Instead, it had independent effects on epithelial organization. Combined genetic and morphological analyses suggested that Pbrm1 inactivation allows cells to sustain Vhl/HIF-dependent proliferation by disrupting tubular architectures that ordinarily restrain this proliferation, resulting in extra-tubular cell accumulation, multilayered epithelia, and tumor formation. Our findings frame a new model for the VHL-PBRM1 interaction that explains loss of epithelial homeostasis through an interaction between discrete effects that drive proliferation and remove structural tissue restraints on that proliferation.

Statement of SignificanceVHL and PBRM1 are frequently co-inactivated in ccRCC. Combining transcriptomic and histological analyses of Vhl and/or Pbrm1-inactivated renal cells in vivo, this study highlights independent effects on transcription and epithelial organization that converge to promote sustained proliferation and dysplasia. The work illuminates how tissue homeostasis is disrupted by oncogenic co-operation.
]]></description>
<dc:creator>Kurlekar, S.</dc:creator>
<dc:creator>Lima, J. D. C. C.</dc:creator>
<dc:creator>Kupfer, N.</dc:creator>
<dc:creator>Pugh, C. W.</dc:creator>
<dc:creator>Mole, D. R.</dc:creator>
<dc:creator>Adam, J.</dc:creator>
<dc:creator>Ratcliffe, P. J.</dc:creator>
<dc:date>2026-02-20</dc:date>
<dc:identifier>doi:10.64898/2026.02.18.706657</dc:identifier>
<dc:title><![CDATA[Discrete genetic effects of VHL and PBRM1 inactivation co-operate to disrupt epithelial homeostasis and promote ccRCC]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.02.12.705585v1?rss=1">
<title>
<![CDATA[
Membrane lipid composition and endocytosis modulate Wingless release from secreting cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.02.12.705585v1?rss=1"
</link>
<description><![CDATA[
Wnts are secreted signalling molecules that regulate development and adult homeostasis. Most Wnts carry a lipid moiety that must be shielded from the aqueous environment. In the secretory pathway, this is achieved by a hydrophobic tunnel in Wntless, a multipass transmembrane protein. However, the Wnt lipid moiety must be released from Wntless before Wnts can engage with Frizzled receptors on receiving cells. Here we address the cell biological basis of Wnt-Wntless dissociation, using as a model the secretion of Drosophila Wingless in wing primordia. Super-resolution microscopy shows that Wingless first reaches the apical surface before being re-internalized to reach, without Wntless, specialized Rab7/Rab4-positive endosomes. From there Wingless traffics to the basolateral membrane where it can engage with glypicans to form a basolateral gradient. Acute inhibition of endocytosis, either with a temperature-sensitive dynamin mutant or a novel optogenetic means of inhibiting clathrin, leads to apical Wingless release in abnormal punctae devoid of Wntless, suggesting that Wingless-Wntless dissociation commences at the apical surface, perhaps because of a distinct lipid composition there. Indeed, similar looking punctae are produced upon genetic abrogation of the ceramide synthase Schlank, specifically in Wingless-producing cells. These punctae resemble insoluble aggregates that form in vitro upon detergent removal. Accordingly, punctae formation can be prevented by shielding the Wingless lipid, in vivo with excess Dally-like protein (Dlp) or in vitro with liposomes. Our results show that membrane lipid composition modulates the orderly transfer of Wingless lipid from Wntless to the inner endosomal surface thus preventing aggregation and ensuring seamless secretion in the basolateral space.
]]></description>
<dc:creator>Alvarez-Rodrigo, I.</dc:creator>
<dc:creator>Alexandre, C.</dc:creator>
<dc:creator>Lebarbachon, C.</dc:creator>
<dc:creator>Borg, A.</dc:creator>
<dc:creator>Finley, H.</dc:creator>
<dc:creator>Arahouan, I.</dc:creator>
<dc:creator>Zhao, Y.</dc:creator>
<dc:creator>Di Pietro, F.</dc:creator>
<dc:creator>Kjaer, S.</dc:creator>
<dc:creator>Booth, P.</dc:creator>
<dc:creator>Bauer, R.</dc:creator>
<dc:creator>Jones, E. Y.</dc:creator>
<dc:creator>Bellaiche, Y.</dc:creator>
<dc:creator>Vincent, J.-P.</dc:creator>
<dc:creator>Willnow, D.</dc:creator>
<dc:date>2026-02-18</dc:date>
<dc:identifier>doi:10.64898/2026.02.12.705585</dc:identifier>
<dc:title><![CDATA[Membrane lipid composition and endocytosis modulate Wingless release from secreting cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.02.18.706528v1?rss=1">
<title>
<![CDATA[
Microglia Rank signaling regulates GnRH function and the Hypothalamic-Pituitary-Gonadal axis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.02.18.706528v1?rss=1"
</link>
<description><![CDATA[
The hypothalamic-pituitary-gonadal axis (HPG) controls pubertal development, sexual maturation, and fertility. We hereby demonstrate a new role of hypothalamic microglia in controlling the HPG axis through Rank signaling, a pathway known for its role in bone and mammary gland biology. Whole-body and microglia Rank depletion leads to hypogonadotropic hypogonadism (HH) resulting from an alteration in gonadotropin-releasing hormone (GnRH) function. In addition, we identify rare gene variants of RANK in patients with HH. Transcriptional profiling upon pubertal Rank loss reveals defective microglia activation and morphological alterations in the median eminence (ME), decreasing the contacts and engulfment of GnRH terminal projections and impairing GnRH responses to kisspeptin. Overall, our data uncovers the crucial role of microglia in regulating GnRH function through Rank signaling, with implications for reproductive maturation and fertility.

One-Sentence SummaryMicroglia regulates GnRH function through RANK signaling
]]></description>
<dc:creator>Collado-Sole, A.</dc:creator>
<dc:creator>Borjini, N.</dc:creator>
<dc:creator>Zhai, J.</dc:creator>
<dc:creator>Soria-Alcaide, G.</dc:creator>
<dc:creator>Folgueira, C.</dc:creator>
<dc:creator>Ruiz-Pino, F.</dc:creator>
<dc:creator>Garcia Vilela, C.</dc:creator>
<dc:creator>Lopez, V.</dc:creator>
<dc:creator>Zouaghi, Y.</dc:creator>
<dc:creator>Jacobs, A.</dc:creator>
<dc:creator>Mora-Romero, B.</dc:creator>
<dc:creator>Barranco, A.</dc:creator>
<dc:creator>Yoldi, G.</dc:creator>
<dc:creator>Rizzoti, K.</dc:creator>
<dc:creator>Sabio, G.</dc:creator>
<dc:creator>Perez-Chacon, G.</dc:creator>
<dc:creator>G. Santamaria, P.</dc:creator>
<dc:creator>Antonio Esteban, J.</dc:creator>
<dc:creator>Pascual, A.</dc:creator>
<dc:creator>Fernandez-Chacon, R.</dc:creator>
<dc:creator>Tena-Sempere, M.</dc:creator>
<dc:creator>Pitteloud, N.</dc:creator>
<dc:creator>Gonzalez-Suarez, E.</dc:creator>
<dc:date>2026-02-18</dc:date>
<dc:identifier>doi:10.64898/2026.02.18.706528</dc:identifier>
<dc:title><![CDATA[Microglia Rank signaling regulates GnRH function and the Hypothalamic-Pituitary-Gonadal axis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.02.25.707914v1?rss=1">
<title>
<![CDATA[
Co-option of a mouse-specific retrotransposon rewires Ash2l isoform usage to prime developmental promoters 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.02.25.707914v1?rss=1"
</link>
<description><![CDATA[
Transcription start site (TSS) selection diversifies the transcriptome and proteome, yet how alternative TSSs regulate development remains unclear. We show that the chromatin regulator ASH2L undergoes developmentally regulated alternative TSS switching in differentiating mouse cells, generating distinct mRNA and protein isoforms: a full-length ASH2L and a truncated form lacking an intrinsically disordered region (IDR). While both ASH2L isoforms are conserved across mammals, their regulation in mouse pluripotent stem cells is uniquely driven by a mouse-specific retrotransposon. This element suppresses transcription from the downstream TSS through a transcription interference mechanism involving SETD2-directed histone H3 lysine 36 methylation. The resulting stem cell-specific truncated ASH2L isoform primes developmental gene promoters for histone H3 lysine 4 methylation, establishing a chromatin state required for embryogenesis and motor neuron differentiation. These findings show that co-option of a mouse-specific retrotransposon rewires Ash2l TSS and protein isoform usage to control developmental timing and cell fate decisions.
]]></description>
<dc:creator>Elgood Hunt, E.</dc:creator>
<dc:creator>Vivori, C.</dc:creator>
<dc:creator>Mitter, R.</dc:creator>
<dc:creator>Hannah Johnkingsly Jebaraj, J.</dc:creator>
<dc:creator>Agnadottir, V.</dc:creator>
<dc:creator>Delas, J.</dc:creator>
<dc:creator>Serna Morales, E.</dc:creator>
<dc:creator>Frith, T.</dc:creator>
<dc:creator>Skehel, M.</dc:creator>
<dc:creator>Elosegui-Artola, A.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>van Werven, F.</dc:creator>
<dc:date>2026-02-26</dc:date>
<dc:identifier>doi:10.64898/2026.02.25.707914</dc:identifier>
<dc:title><![CDATA[Co-option of a mouse-specific retrotransposon rewires Ash2l isoform usage to prime developmental promoters]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.02.25.707913v1?rss=1">
<title>
<![CDATA[
An injectable soft implant for long-acting, reversible, ultra-stable release of therapeutics 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.02.25.707913v1?rss=1"
</link>
<description><![CDATA[
Providing long-term (>6 months) zero-order drug release from easily administered formulations is a key challenge in improving patient adherence and facilitating access. Herein, we report the design and development of an injectable, biodegradable, long-acting polymeric microparticle-embedded hydrogel platform for prolonged, zero-order release of therapeutics. This "soft implant" is injectable for ease of administration and can be retrieved via a small incision, allowing for discontinuation of therapy if desired. Central to the platform are surface-eroding poly(orthoester) (POE) microparticles, which were molecularly tailored to tune zero-order drug release across a wide range of timeframes. We demonstrate the clinical potential of the "soft implant" using levonorgestrel, a contraceptive agent requiring sustained dosing. In vitro, we observed zero-order release for 300 days, projected for >12 months, with behavior consistent with surface erosion further supported through Raman chemical mapping. In vivo studies confirmed zero-order release for six months, projected to 12 months, from a subcutaneous injection in rats. We envision that our platform could transform therapies that require long-term, regular drug dosing, significantly improving compliance and therapy outcomes.
]]></description>
<dc:creator>Stevens, M. M.</dc:creator>
<dc:creator>Kütahya, C.</dc:creator>
<dc:creator>Panariello, L.</dc:creator>
<dc:creator>Najer, A.</dc:creator>
<dc:creator>Rizou, T.</dc:creator>
<dc:creator>Shamsabadi, A.</dc:creator>
<dc:creator>Brachi, G.</dc:creator>
<dc:creator>Peeler, D. J.</dc:creator>
<dc:creator>Zharova, L.</dc:creator>
<dc:creator>Fernandez Debets, T. F. F.</dc:creator>
<dc:creator>Peschke, P.</dc:creator>
<dc:creator>Constantinou, A. P.</dc:creator>
<dc:creator>Xie, R.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:creator>Burdis, R.</dc:creator>
<dc:creator>Suarez-Bonnet, A.</dc:creator>
<dc:creator>Cihova, M.</dc:creator>
<dc:creator>Yeow, J.</dc:creator>
<dc:creator>Schaufelberger, F.</dc:creator>
<dc:creator>Malanchi, I.</dc:creator>
<dc:date>2026-02-26</dc:date>
<dc:identifier>doi:10.64898/2026.02.25.707913</dc:identifier>
<dc:title><![CDATA[An injectable soft implant for long-acting, reversible, ultra-stable release of therapeutics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.04.01.646648v1?rss=1">
<title>
<![CDATA[
iNeurons are sweet, maybe too sweet? Exploring the impact of media composition on PINK1-dependent mitophagy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.04.01.646648v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease associated proteins PINK1 and Parkin collaboratively regulate stress-induced mitophagy. While in vitro human neuronal cultures are valuable for studying the roles of PINK1 and Parkin in a disease-relevant context, the impact of culture conditions on these processes remains largely underexplored. Here, it is shown that human induced neurons (iNeurons) cultured in N2B27 and BrainPhys medium exhibit distinct PINK1-Parkin dependent mitophagy phenotypes. Specifically, BrainPhys-cultured iNeurons show greater resistance to PINK1-dependent mitophagy initiation, linked to a reduction in glucose availability and reduced PINK1 protein availabilities, leading to decreases in stress-induced and basal mitophagy fluxes. These findings highlight the critical impact of culture conditions on mitophagy dynamics and emphasise the need to account for media-specific differences when using in vitro models to investigate mitophagy mechanisms in human neurons.
]]></description>
<dc:creator>O'Callaghan, B.</dc:creator>
<dc:creator>Melandri, D.</dc:creator>
<dc:creator>Soltic, D.</dc:creator>
<dc:creator>Cosker, K.</dc:creator>
<dc:creator>Soutar, M. P. M.</dc:creator>
<dc:creator>Plun-Favreau, H.</dc:creator>
<dc:date>2025-04-03</dc:date>
<dc:identifier>doi:10.1101/2025.04.01.646648</dc:identifier>
<dc:title><![CDATA[iNeurons are sweet, maybe too sweet? Exploring the impact of media composition on PINK1-dependent mitophagy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.02.24.707647v1?rss=1">
<title>
<![CDATA[
Myc inhibition triggers GM-CSF-driven regression of pancreatic tumours 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.02.24.707647v1?rss=1"
</link>
<description><![CDATA[
The complex aetiology of pancreatic ductal adenocarcinoma (PDAC), together with its desmoplastic and hypoxic microenvironment, hinders effective treatment and contributes to its rapid lethality1. Using a reversibly switchable genetic mouse model that closely recapitulates human PDAC phenotype progression, we previously showed that selective deactivation of oncogenic Myc in PDAC epithelial cells triggers rapid disassembly of advanced PDACs2, both tumor cells and their associated immune microenvironment. Here, leveraging spatiotemporal genomics and multiplex immune profiling we determine the mechanism underpinning this regression programme and identify granulocyte-macrophage colony-stimulating factor (GM-CSF) as its key instructive cytokine, transiently released by pancreatic ductal epithelial cells rapidly following Myc inactivation, that initiates tumour regression. We further demonstrate that type 1 conventional dendritic cells (cDC1s) act as critical effectors in Myc-OFF GM-CSF-driven tumour regression. Both antibody-mediated blockade of GM-CSF and genetic ablation of cDC1s via Batf3 knockout bone marrow transplantation prevent PDAC regression. Conversely, transient systemic administration of recombinant GM-CSF to PDAC-bearing mice promotes rapid cDC1 infiltration and induces marked tumour regression in the sustained presence of Myc activity. Together, these findings reveal that PDAC regression induced by Myc de-activation is mediated by a latent morphogenic programme that is initiated by transient release of GM-CSF from tumour cells. This regression programme is rapid, tissue-specific, involves both neoplastic cells and attendant desmoplastic stroma, is reliant on innate immunity and provides a novel framework for therapeutic intervention in PDAC.
]]></description>
<dc:creator>Campos, T.</dc:creator>
<dc:creator>Kortlever, R. M.</dc:creator>
<dc:creator>Sodir, N. M.</dc:creator>
<dc:creator>Buck, M. D.</dc:creator>
<dc:creator>Stockis, J.</dc:creator>
<dc:creator>Parker, J.</dc:creator>
<dc:creator>Lam, C. M.</dc:creator>
<dc:creator>Patino-Mercau, J. R.</dc:creator>
<dc:creator>Perfetto, A.</dc:creator>
<dc:creator>Edwards, A.</dc:creator>
<dc:creator>Stefan, B.</dc:creator>
<dc:creator>Halim, T. Y.</dc:creator>
<dc:creator>Gerard, E. I.</dc:creator>
<dc:date>2026-02-25</dc:date>
<dc:identifier>doi:10.64898/2026.02.24.707647</dc:identifier>
<dc:title><![CDATA[Myc inhibition triggers GM-CSF-driven regression of pancreatic tumours]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.02.27.708290v1?rss=1">
<title>
<![CDATA[
A stable subgenomic reporter coronavirus enables transcriptional profiling of bystander cells. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.02.27.708290v1?rss=1"
</link>
<description><![CDATA[
Insertion of fluorescent reporter genes into viral genomes is a powerful tool for monitoring infection. In coronaviruses, this is commonly achieved by replacing accessory open reading frames, thereby deleting endogenous gene functions. An alternative strategy is to manipulate viral RNA synthesis by inserting copies of the viral transcription regulatory sequence (TRS) which drive the transcription of viral subgenomic RNAs. However, coronavirus transcription is tightly regulated, and these modifications frequently disrupt native subgenomic RNA synthesis and attenuate viral growth. Here, we describe a reporter coronavirus that overcomes these limitations. Using human coronavirus (HCoV)-OC43 as a model system, we inserted an mNeonGreen reporter between the Spike and ORF5 coding regions, engineering the TRS and surrounding sequence to minimise off-target effects to transcription. This virus is genetically stable, with wildtype growth kinetics and unaltered subgenomic RNA transcriptional ratios. We developed a flexible reverse genetics system, which allows rapid cloning and virus recovery, supported by optimised HCoV-OC43 culture conditions, for high-titre stock generation, and validated analytical reagents. Our reporter virus enabled sensitive detection and isolation of infected cells, facilitating transcriptomic analyses that distinguish host responses in infected and bystander populations. We found that transcriptional responses to infection of cells in culture were predominantly inflammatory, rather than interferon-mediated, and that bystander cells upregulated pathways associated with cytokine response signalling and cell-cell contact sensing. Together, these tools expand the experimental utility of HCoV-OC43, an important seasonal respiratory pathogen and low containment model for betacoronavirus biology.
]]></description>
<dc:creator>Gilbride, C.</dc:creator>
<dc:creator>Hemsley-Taylor, J.</dc:creator>
<dc:creator>Nunes, C.</dc:creator>
<dc:creator>Penn, R.</dc:creator>
<dc:creator>Boot, J.</dc:creator>
<dc:creator>Pieris, N.</dc:creator>
<dc:creator>Tripathy, R.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Hutchinson, M.</dc:creator>
<dc:creator>Platt, O. K.</dc:creator>
<dc:creator>Ulferts, R.</dc:creator>
<dc:creator>Mitter, R.</dc:creator>
<dc:creator>Strom, M.</dc:creator>
<dc:creator>Santos, N. B.</dc:creator>
<dc:creator>Bauer, D. L.</dc:creator>
<dc:creator>Mears, H. V.</dc:creator>
<dc:date>2026-03-03</dc:date>
<dc:identifier>doi:10.64898/2026.02.27.708290</dc:identifier>
<dc:title><![CDATA[A stable subgenomic reporter coronavirus enables transcriptional profiling of bystander cells.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.03.10.710544v1?rss=1">
<title>
<![CDATA[
Contribution of cytotoxic CD8 T cells, neutrophils and type 1 interferon signaling to hyperinflammatory pathology in HIV associated TB meningitis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.03.10.710544v1?rss=1"
</link>
<description><![CDATA[
Immune dysregulation contributes to death and disability in tuberculous meningitis. People living with HIV have the least evidence that anti-inflammatory therapy improves the poor outcome. Improving therapy relies on a more refined understanding of the host immune response. Single-cell RNA sequencing of 188,983 CSF cells from 25 adults with HIV-associated TBM revealed a predominance of cytotoxic CD8 T cells with low cytokine expression. In microbiologically-confirmed TBM, there was greater cytotoxicity in T, NK and {gamma}{delta} cells, and higher type 1 interferon stimulation in T and B lymphocytes. Neutrophils expressed markers suggesting heightened cytokine stimulation, enhanced effector function, and IL-8-mediated neutrophil recruitment. In a longitudinal cohort, type 1 interferon signaling increased in blood and CSF following treatment initiation. Overall, findings indicate a hyper-inflammatory immune response in the CSF of HIV-associated TBM patients characterised by an accumulation of granzyme-rich cytotoxic CD8 T cells, highly activated neutrophils and host-detrimental type 1 interferon signaling.
]]></description>
<dc:creator>Barnacle, J. R.</dc:creator>
<dc:creator>Bangani, N.</dc:creator>
<dc:creator>Slawinski, H.</dc:creator>
<dc:creator>Barrington, C.</dc:creator>
<dc:creator>Wilkinson, K. A.</dc:creator>
<dc:creator>Stek, C. J.</dc:creator>
<dc:creator>Lai, R.</dc:creator>
<dc:creator>Meintjes, G.</dc:creator>
<dc:creator>Robertson, B. D.</dc:creator>
<dc:creator>Gengenbacher, M.</dc:creator>
<dc:creator>Davis, A. G.</dc:creator>
<dc:creator>Barber, D. L.</dc:creator>
<dc:creator>O'Garra, A.</dc:creator>
<dc:creator>Wilkinson, R. J.</dc:creator>
<dc:date>2026-03-10</dc:date>
<dc:identifier>doi:10.64898/2026.03.10.710544</dc:identifier>
<dc:title><![CDATA[Contribution of cytotoxic CD8 T cells, neutrophils and type 1 interferon signaling to hyperinflammatory pathology in HIV associated TB meningitis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.03.07.710300v1?rss=1">
<title>
<![CDATA[
Meiotic cohesin Rec8 imposes fitness costs on fission yeast gametesfavoring the evolution of parental bias in gene expression 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.03.07.710300v1?rss=1"
</link>
<description><![CDATA[
Differences between partner gametes, which evolved repeatedly in eukaryotes, can contribute to the evolution of the sexes, sexual selection and non-Mendelian inheritance1- 4. Yet, the empirical evidence for how functional asymmetries arise between initially equivalent gametes is limited. Here, we combine theoretical and experimental approaches in the fission yeast Schizosaccharomyces pombe to show how selective pressures acting concurrently on gametes and zygotes drive the evolution of gamete differences. We find that despite being morphologically identical, P- and M-type partner gametes invest asymmetrically in zygotic development by contributing different amounts of conserved meiotic cohesins. P-gametes preferentially produce thve Rec85 cohesin that increases zygotic fitness but reduces gamete viability, revealing a trade-off between reproductive success and gamete survival. We demonstrate that this asymmetry is mediated by partner-specific communication and model its evolutionary dynamics using empirically determined parameters. Our results support classical theoretical predictions for the evolution of gamete differences and provide a mechanistic understanding of how molecular asymmetries between partners can originate from opposing selection pressures acting in species that lack morphologically distinct gametes.
]]></description>
<dc:creator>Martins, C.</dc:creator>
<dc:creator>Booth, H.</dc:creator>
<dc:creator>Salat Canela, C.</dc:creator>
<dc:creator>Hadjivasiliou, Z.</dc:creator>
<dc:creator>Vjestica, A.</dc:creator>
<dc:date>2026-03-08</dc:date>
<dc:identifier>doi:10.64898/2026.03.07.710300</dc:identifier>
<dc:title><![CDATA[Meiotic cohesin Rec8 imposes fitness costs on fission yeast gametesfavoring the evolution of parental bias in gene expression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.03.09.707383v1?rss=1">
<title>
<![CDATA[
A DNA foundation model predicts osteoporosis risk genes without proximity bias 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.03.09.707383v1?rss=1"
</link>
<description><![CDATA[
Targets supported by human genetic associations are more than twice as likely to progress from clinical development to approval. Genome-wide association studies are the largest source of genetic evidence for disease risk but linking non-coding variants to effector genes remains a significant barrier to identifying causal targets. Current gene-mapping approaches suffer from proximity bias, largely ignoring distal genes. Here we introduce Rosalind, a DNA foundation model fine-tuned on human genetic variation from GTEx, that directly predicts variant-gene regulatory relationships from sequence without relying on nearest-gene heuristics. We demonstrate Rosalinds accuracy through extensive benchmarking, apply it to multiple complex traits to establish broad utility, and provide experimental validation in osteoporosis using a translational osteoblast assay. We demonstrate that genes distal to osteoporosis risk variants were significantly more likely to alter a bone formation phenotype than nearest genes. Together, these results highlight deep learning-based regulatory models as a general and scalable framework for translating novel genetic insights to drug discovery.
]]></description>
<dc:creator>Regep, C.</dc:creator>
<dc:creator>Kapourani, C.-A.</dc:creator>
<dc:creator>Sofyali, E.</dc:creator>
<dc:creator>Dobrowolska, A.</dc:creator>
<dc:creator>Loukas, G.</dc:creator>
<dc:creator>Anighoro, A.</dc:creator>
<dc:creator>Canale, E.</dc:creator>
<dc:creator>Gross, T.</dc:creator>
<dc:creator>Licciardello, M.</dc:creator>
<dc:creator>Gupta, R.</dc:creator>
<dc:creator>Maciuca, S.</dc:creator>
<dc:creator>Desai, T.</dc:creator>
<dc:creator>Del Vecchio, A.</dc:creator>
<dc:creator>Field, C.</dc:creator>
<dc:creator>Gemayel, K.</dc:creator>
<dc:creator>Javer, A.</dc:creator>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Tsujikawa, R.</dc:creator>
<dc:creator>Inoue, F.</dc:creator>
<dc:creator>Hessel, E.</dc:creator>
<dc:creator>Taylor-King, J.</dc:creator>
<dc:creator>Whittaker, J.</dc:creator>
<dc:creator>Roblin, D.</dc:creator>
<dc:creator>McIntyre, R.</dc:creator>
<dc:creator>Edwards, L.</dc:creator>
<dc:date>2026-03-12</dc:date>
<dc:identifier>doi:10.64898/2026.03.09.707383</dc:identifier>
<dc:title><![CDATA[A DNA foundation model predicts osteoporosis risk genes without proximity bias]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.03.10.698104v1?rss=1">
<title>
<![CDATA[
The phosphoproteomic landscape of the neurological manifestations in tuberous sclerosis complex 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.03.10.698104v1?rss=1"
</link>
<description><![CDATA[
Tuberous sclerosis complex (TSC) is a rare disease caused by mutations in TSC1 and TSC2, resulting in activation of mechanistic target of rapamycin complex 1 (mTORC1). Neurological manifestations in TSC patients include epilepsy, autism and intellectual disability. Two types of brain lesions, cortical tubers and subependymal giant cell astrocytomas (SEGAs), cause the majority of neurological manifestations in TSC. We have limited understanding of the molecular changes that occur in tubers and SEGAs and how these contribute to disease pathogenesis. To investigate this, we performed proteomic and phosphoproteomic analysis of TSC patient tuber and SEGA tissue. Tubers showed evidence of alterations in mitochondrial respiration, cytoskeleton organisation and neuronal function. However, we were unable to detect mTORC1 activation in tubers, likely due to the small number of cells with complete inactivation of TSC1 or TSC2. By contrast, SEGAs showed evidence of strong mTORC1 activation and large-scale changes in the proteome and phosphoproteome. SEGAs exhibited increased expression of ribosomal proteins and activation of a neuroinflammatory response. Phosphoproteomics identified 6060 phosphosites within 2154 proteins increased in SEGAs. Phosphorylation of multiple proteins involved in RNA-metabolism, including mRNA splicing, were increased in SEGAs. Consistent with this, we found evidence of extensive alterations in mRNA transcript splicing in SEGA tissue. These data greatly expand the repertoire of known mTORC1 target proteins in the human brain and reveal large-scale mis-regulation of mRNA splicing in SEGAs in TSC.
]]></description>
<dc:creator>Girodengo, M.</dc:creator>
<dc:creator>Mihaylov, S. R.</dc:creator>
<dc:creator>Klonowska, K.</dc:creator>
<dc:creator>Mantoan Ritter, L.</dc:creator>
<dc:creator>Flynn, H. R.</dc:creator>
<dc:creator>Skehel, M.</dc:creator>
<dc:creator>Bou Farhat, E.</dc:creator>
<dc:creator>Aronica, E.</dc:creator>
<dc:creator>White, M. A.</dc:creator>
<dc:creator>Kwiatkowski, D.</dc:creator>
<dc:creator>Ultanir, S. K.</dc:creator>
<dc:creator>Bateman, J. M.</dc:creator>
<dc:date>2026-03-12</dc:date>
<dc:identifier>doi:10.64898/2026.03.10.698104</dc:identifier>
<dc:title><![CDATA[The phosphoproteomic landscape of the neurological manifestations in tuberous sclerosis complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.03.09.710501v1?rss=1">
<title>
<![CDATA[
Inferring state-dependent functional circuit motifs using higher-order interactions analysis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.03.09.710501v1?rss=1"
</link>
<description><![CDATA[
Analysing higher-order interactions among simultaneously recorded neurons can provide crucial insights into neural network dynamics. Recent technological advances have enabled large-scale, long-term neuronal recordings, but analysis of such datasets often relies on simpler statistics due to computational and statistical challenges in assessing higher-order interactions. Here, we developed CHOIR, an efficient and reliable method for calculating higher-order interactions from large-scale neuronal recordings. We then used the inferred HOIs to uncover the underlying functional connectivity, differentiating between connectivity motifs in the space of pairwise and triplet-wise interactions. We found that this approach could successfully distinguish stationary and running states, sleep and awake states, and neuronal ensembles with distinct activity patterns in mice. Furthermore, we identified potential circuit architectures underlying different higher-order interactions, which we confirmed through simulations of large-scale spiking networks with specific subnetwork connectivity. Applying CHOIR to a causal manipulation dataset further confirmed the role of lateral inhibition, a key inhibitory motif, in generating specific HOI patterns. Our work provides a systematic analysis of higher-order interactions in diverse datasets and suggests that HOIs can reveal circuit motifs underlying neural dynamics across brain areas and brain states.
]]></description>
<dc:creator>Rashid Shomali, S.</dc:creator>
<dc:creator>Rasuli, S. N.</dc:creator>
<dc:creator>Shimazaki, H.</dc:creator>
<dc:creator>Sadeh, S.</dc:creator>
<dc:date>2026-03-11</dc:date>
<dc:identifier>doi:10.64898/2026.03.09.710501</dc:identifier>
<dc:title><![CDATA[Inferring state-dependent functional circuit motifs using higher-order interactions analysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.03.22.713469v1?rss=1">
<title>
<![CDATA[
Myofibers drive postnatal myonuclear accretion through Arp2/3-dependent membrane protrusions 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.03.22.713469v1?rss=1"
</link>
<description><![CDATA[
Skeletal muscle postnatal growth depends on myoblast fusion with pre-existing myofibers, but whether myofibers actively regulate this process remains unclear. The Arp2/3 complex, which nucleates branched actin networks, is required for myoblast fusion during embryogenesis, but its role in postnatal myofibers is unknown. Here, we investigated the role of the Arp2/3 complex in myofibers using an inducible skeletal muscle-specific Arpc4 knockout model. We find that loss of Arp2/3 in myofibers impairs myonuclear accretion, resulting in smaller myofibers and reduced muscle strength. Satellite cells are still activated and able to differentiate but accumulate around Arpc4-deficient myofibers rather than fusing, pointing to a myofiber-intrinsic defect. In co-culture, myofibers form long-lived membrane protrusions enriched in Arp2/3 at myoblast contact sites, and optogenetic activation of protrusion in myofibers induced myoblast fusion. Genetic depletion of Arpc4 in myofibers markedly reduces membrane protrusions both in vitro and in vivo. These findings unveil a fundamental role for the Arp2/3 complex in postnatal muscle growth, and reveal a mechanism by which myofibers actively extend membrane protrusions to drive myoblast fusion. This repositions myofibers as active fusogenic partners and cell-autonomous drivers of their own nuclear accretion.
]]></description>
<dc:creator>Sequeira, C.</dc:creator>
<dc:creator>Di Francescantonio, S.</dc:creator>
<dc:creator>Motta, G.</dc:creator>
<dc:creator>Kogata, N.</dc:creator>
<dc:creator>Way, M.</dc:creator>
<dc:creator>Gomes, E. R.</dc:creator>
<dc:date>2026-03-23</dc:date>
<dc:identifier>doi:10.64898/2026.03.22.713469</dc:identifier>
<dc:title><![CDATA[Myofibers drive postnatal myonuclear accretion through Arp2/3-dependent membrane protrusions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.03.28.714971v1?rss=1">
<title>
<![CDATA[
The antipsychotic drug clozapine suppresses autoimmunity driving psychosis-like behavior in mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.03.28.714971v1?rss=1"
</link>
<description><![CDATA[
Antipsychotic drugs are the first-line treatment for psychosis yet their mechanism of action remains poorly understood, largely due to the challenge to faithfully model psychosis preclinically. Here, we focus on the emerging concept that psychosis can be caused by brain autoimmunity and present a novel mouse model of anti-N-methyl-D-aspartate-receptor (anti-NMDAR) encephalitis, a condition that manifests with psychosis and autoanti-bodies against the NMDAR. We devised a new mRNA-based approach to immunize mice against the NMDAR. Immunized mice developed psychosis-like behaviors that were caused by anti-NMDAR autoantibodies leading to phagocytosis of NMDARs by brain microglia. The antipsychotic drug clozapine rescued psychosis-like behaviors and, remarkably, reduced anti-NMDAR autoantibody levels and antibody-mediated phagocytosis of NMDARs. The immunomodulatory effects of clozapine were confirmed in a mouse model of systemic lupus erythematosus. Our results demonstrate that clozapine suppresses autoimmunity driving psychosis-like behaviors, raising the possibility that immunomodulation contributes to antipsychotic drug action.

HIGHLIGHTSO_LImRNA immunization against the NMDAR induces psychosis-like behavior in mice
C_LIO_LIAnti-NMDAR autoantibodies are sufficient for psychosis-like behavior
C_LIO_LIMicroglial phagocytosis of NMDARs mediates psychosis-like behavior induced by anti-NMDAR autoanti-bodies.
C_LIO_LIClozapine reduces anti-NMDAR autoantibodies, microglial phagocytosis and psychosis-like behavior, consistent with immunomodulation as a potential mechanism of antipsychotic drug action.
C_LI
]]></description>
<dc:creator>He, L.</dc:creator>
<dc:creator>Feldman, H.</dc:creator>
<dc:creator>Nguyen, T.</dc:creator>
<dc:creator>Bosc, M.</dc:creator>
<dc:creator>Polisetty, V.</dc:creator>
<dc:creator>Kriel, O.</dc:creator>
<dc:creator>Landwehr, A.</dc:creator>
<dc:creator>Borg, A.</dc:creator>
<dc:creator>Subtil, F. T.</dc:creator>
<dc:creator>Khakpour, M.</dc:creator>
<dc:creator>Zhou, J.</dc:creator>
<dc:creator>Kjaer, S.</dc:creator>
<dc:creator>MacCabe, J.</dc:creator>
<dc:creator>Pollak, T. A.</dc:creator>
<dc:creator>Tremblay, M.-E.</dc:creator>
<dc:creator>Vinuesa, C. G.</dc:creator>
<dc:creator>Hayday, A.</dc:creator>
<dc:creator>Schmack, K.</dc:creator>
<dc:date>2026-03-31</dc:date>
<dc:identifier>doi:10.64898/2026.03.28.714971</dc:identifier>
<dc:title><![CDATA[The antipsychotic drug clozapine suppresses autoimmunity driving psychosis-like behavior in mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.03.27.714575v1?rss=1">
<title>
<![CDATA[
Natural statistics of host odours predict species-specific olfactory behaviours in Drosophilids 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.03.27.714575v1?rss=1"
</link>
<description><![CDATA[
Animals rely on olfaction to locate food, mates, and suitable habitats, yet natural odour environments contain thousands of volatile molecules, creating a high-dimensional sensory problem for both nervous systems and the researchers who study them 1-5. For example, a banana emits around 100 individual volatiles4,6. It remains unclear which components of complex odour blends animals have evolved to use as behavioural cues. Here, combining fieldwork, chemical and behavioural analyses, we show across multiple Drosophila species that behaviourally relevant cues can be predicted directly from the statistical structure of natural odour environments. Animals preferentially respond to components that are most distinctive within their natural host odour blends, and therefore most ecologically informative. These cues can be either major or minor blend components. Our results indicate that host-guided olfactory behaviours have evolved to exploit the statistical structure of natural odour environments by selectively targeting the most informative features of odour blends.
]]></description>
<dc:creator>Gong, H.</dc:creator>
<dc:creator>Ziolkowska, Z.</dc:creator>
<dc:creator>Khallaf, M. A.</dc:creator>
<dc:creator>Pop, S.</dc:creator>
<dc:creator>Ayrton, O.</dc:creator>
<dc:creator>Cano-Ferrer, X.</dc:creator>
<dc:creator>MacRae, J.</dc:creator>
<dc:creator>Knaden, M.</dc:creator>
<dc:creator>Arguello, R.</dc:creator>
<dc:creator>Prieto-Godino, L. L.</dc:creator>
<dc:date>2026-03-31</dc:date>
<dc:identifier>doi:10.64898/2026.03.27.714575</dc:identifier>
<dc:title><![CDATA[Natural statistics of host odours predict species-specific olfactory behaviours in Drosophilids]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.03.26.714455v1?rss=1">
<title>
<![CDATA[
A genome-wide atlas of meiotic recombination intermediates reveals distinct modes of DNA repair that direct crossovers away from transcriptionally marked genes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.03.26.714455v1?rss=1"
</link>
<description><![CDATA[
Crossovers are essential for accurate chromosome segregation in meiosis. Yet the programmed DNA double-strand breaks that initiate them frequently occur in genes and pose a risk to transcription required for gametogenesis. How meiotic cells reconcile these competing demands has remained unclear. Here, we generate a genome-wide in vivo atlas of meiotic recombination intermediates across [~]42,000 hotspots by mapping repair proteins BLM, HFM1, and RPA in wild-type and genome-engineered mutant mouse testes. These maps reveal two distinct modes of break repair: a fast-resolving class with short-lived intermediates that are repaired predominantly as non-crossovers, and a slower class with persistent intermediates that give rise to nearly all crossovers. Fast-resolving hotspots occur almost exclusively within a deeply conserved set of [~]4,500 genes marked by structural and chromatin features established during an early stage of meiotic transcription. This transcriptional memory predicts repair fate with high accuracy across mouse subspecies and sexes. Across widely diverged mammals, including humans and cattle, orthologous genes show similar crossover suppression. Our findings reveal an early bifurcation between crossover and non-crossover repair that is governed by the transcriptional context of meiotic breaks. Together, they establish an evolutionarily conserved principle in which crossovers are directed away from transcriptionally important genes, thereby safeguarding gene function and shaping their evolution.
]]></description>
<dc:creator>Henfrey, C.</dc:creator>
<dc:creator>Print, E.</dc:creator>
<dc:creator>Zhang, G.</dc:creator>
<dc:creator>Hinch, R.</dc:creator>
<dc:creator>Maudlin, I.</dc:creator>
<dc:creator>Moralli, D.</dc:creator>
<dc:creator>Davies, B.</dc:creator>
<dc:creator>Donnelly, P.</dc:creator>
<dc:creator>Hinch, A. G.</dc:creator>
<dc:date>2026-03-28</dc:date>
<dc:identifier>doi:10.64898/2026.03.26.714455</dc:identifier>
<dc:title><![CDATA[A genome-wide atlas of meiotic recombination intermediates reveals distinct modes of DNA repair that direct crossovers away from transcriptionally marked genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.03.24.713949v1?rss=1">
<title>
<![CDATA[
Intravital imaging uncovers remodelling of humanised bone marrow-like niches 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.03.24.713949v1?rss=1"
</link>
<description><![CDATA[
The bone marrow haematopoietic niche is composed of a diverse array of cell types and extracellular matrix components that together support healthy haematopoiesis. However, live imaging of the bone marrow microenvironment is hampered by tissue accessibility limitations. Using intravital imaging through a titanium imaging window, we investigated the dynamics of human haematopoietic cells and mesenchymal stromal cells within an ectopically implanted humanised scaffold in an immunodeficient murine host. These cell populations expand and differentiate over time, accompanied by progressive remodelling of the scaffold. We observe migration of murine endothelial cells into the scaffold, leading to the formation of a vascular network during the initial development of the humanised niche. Subsequently, the dense collagen matrix that makes up the implanted niche is altered and larger gaps form in regions populated by mesenchymal stroma cells. Collectively, our findings demonstrate dynamic remodelling of the extracellular milieu that supports haematopoietic cell development and establish a platform for longitudinal, in vivo investigation of these processes. Altogether, we describe a novel model that aligns with the 3R guiding principles and enables real-time assessment of bone marrow cell dynamics in vivo.

Summary statementRatcliffe and Mian et al. image in vivo dynamics of a bone marrow haematopoietic niche model.
]]></description>
<dc:creator>Ratcliffe, C. D. H.</dc:creator>
<dc:creator>Mian, S. A.</dc:creator>
<dc:creator>Giangreco, G.</dc:creator>
<dc:creator>Le Marois, A.</dc:creator>
<dc:creator>Habel, K.</dc:creator>
<dc:creator>Sahai, E.</dc:creator>
<dc:creator>Bonnet, D.</dc:creator>
<dc:date>2026-03-26</dc:date>
<dc:identifier>doi:10.64898/2026.03.24.713949</dc:identifier>
<dc:title><![CDATA[Intravital imaging uncovers remodelling of humanised bone marrow-like niches]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.03.25.714146v1?rss=1">
<title>
<![CDATA[
Distinct signaling center and progenitor identity dynamics initiate human forebrain patterning 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.03.25.714146v1?rss=1"
</link>
<description><![CDATA[
At which developmental stage does the human brain start to differ from other mammals? We uncover human features of telencephalic patterning as early as the fourth post-conception week, comparing the early mouse and human forebrains using single-cell transcriptomics and 3D spatial multi-transcript imaging. In comparison to mice, the human telencephalon is delayed in ventral SHH signaling, accompanied by a human signature of anterior FGF signaling. These observations correlate with a less-resolved patterning and reduced progenitor diversity along the dorsoventral and anteroposterior axes, as well as a human early neurogenic signature. Our complementary approaches reveal a human divergence in allocation of telencephalic progenitor identities, propelled by temporal and qualitative differences in ventro-anterior signals following neural tube closure in the developing forebrain.
]]></description>
<dc:creator>Azizi, A.</dc:creator>
<dc:creator>Fakhreddine, D.</dc:creator>
<dc:creator>Hamid, F.</dc:creator>
<dc:creator>Messi, Z.</dc:creator>
<dc:creator>Strohbuecker, S.</dc:creator>
<dc:creator>Guillemot, F.</dc:creator>
<dc:creator>Houart, C.</dc:creator>
<dc:date>2026-03-25</dc:date>
<dc:identifier>doi:10.64898/2026.03.25.714146</dc:identifier>
<dc:title><![CDATA[Distinct signaling center and progenitor identity dynamics initiate human forebrain patterning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.03.22.713441v1?rss=1">
<title>
<![CDATA[
Structures and molecular mechanisms of RAD54B in modulating homologous recombination 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.03.22.713441v1?rss=1"
</link>
<description><![CDATA[
Genome stability is essential for cellular viability yet constantly threatened by endogenous and exogenous DNA-damaging agents. Among these, DNA double-strand breaks (DSBs) are particularly harmful and in S/G2 phases are faithfully repaired through homologous recombination (HR), a high-fidelity pathway utilising homologous sequences in sister chromatin. The RAD51 recombinase forms nucleoprotein filaments on single-stranded DNA (ssDNA) to mediate homology search, strand invasion and subsequent D-loop formation that leads to DNA synthesis and repair. The efficiency of HR depends on precise regulation of RAD51 filament dynamics by accessory factors, including RAD54 and RAD54B, which belong to the SWI2/SNF2-family DNA translocases. While RAD54 is well-characterized, RAD54Bs molecular functions remain poorly understood. Here, we define RAD54Bs role in HR using cryo-electron microscopy, mutagenesis, biochemical and cellular assays. We show that RAD54B stabilizes RAD51-DNA filaments, inhibits RAD51 ATPase activity, and promotes strand invasion, D-loop formation and strand exchange. The N-terminal domain (NTD) alone supports filament stabilization and strand exchange, while the C-terminal ATPase domain is required for D-loop formation. Structural and biochemical analyses reveal three RAD51-interacting sites within the NTD and a unique domain ({beta}-domain) that bridges RAD51 protomers and contacts donor dsDNA. This {beta}-domain also regulates RAD54Bs ATPase activity and higher-order oligomer organization on dsDNA. Cellular assays reveal that the NTD RAD51-interacting sites as well as the {beta}-domain are required for repairing camptothecin-induced DSBs by HR in human cells. Our findings uncover a modular architecture and mechanistic framework for RAD54B function in HR, highlighting its critical role in genome maintenance.

HighlightsO_LIcryoEM structure of RAD54B in complex with RAD51-DNA complex
C_LIO_LIRAD54B uses three sites to interact with RAD51, including a previously unrecognised {beta}-domain that bridges distal RAD51 protomers.
C_LIO_LIThe {beta}-domain plays multiple crucial roles including regulating filament stability, RAD54B ATPase activity and RAD54B higher order assembly on DNA.
C_LIO_LIRAD54B employs a modular mechanism, with the N-terminal region engaing and stabilising RAD51 filaments, capturing of the homologous strands, whereas the ATPase motor domainrequired for homology search and strand invasion.
C_LIO_LIRAD54B N-terminus and {beta}-domain are essential for HR-mediated repair of camptothecin-induced breaks in human cells.
C_LI
]]></description>
<dc:creator>Liang, P.</dc:creator>
<dc:creator>Tye, S.</dc:creator>
<dc:creator>Ertl da Costa, J.</dc:creator>
<dc:creator>Maharshi, N.</dc:creator>
<dc:creator>Argunhan, B.</dc:creator>
<dc:creator>Khulen, L.</dc:creator>
<dc:creator>Battley, M.</dc:creator>
<dc:creator>McCormack, E.</dc:creator>
<dc:creator>Heyer, W.-D.</dc:creator>
<dc:creator>Lobrich, M.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:date>2026-03-24</dc:date>
<dc:identifier>doi:10.64898/2026.03.22.713441</dc:identifier>
<dc:title><![CDATA[Structures and molecular mechanisms of RAD54B in modulating homologous recombination]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.03.23.713733v1?rss=1">
<title>
<![CDATA[
Pro-domain-dependent folding and co-receptor-mediated targeting to optimize an antagonistic TGF-β monomer for gene-based delivery 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.03.23.713733v1?rss=1"
</link>
<description><![CDATA[
Transforming growth factor-beta (TGF-{beta}), a potent promoter of extracellular matrix deposition and suppressor of infiltrating immunity, has arisen as an attractive target for improving outcomes in tissue fibrosis and cancer immune therapy. Despite the promise of TGF-{beta} inhibitors for attenuating the progression of fibrotic disorders or as adjuncts for cancer immunotherapy, current systemically administered inhibitors that target the ligand or receptors have significant on-target liabilities, including cardiotoxicity and development of pre-malignant cutaneous squamous lesions. Recently, an engineered mini monomer of TGF-{beta} (mmTGF-{beta}), which potently and specifically inhibits TGF-{beta} activity, was shown to strongly synergize with checkpoint inhibitors to suppress cancer progression in an aggressive model of melanoma when genetically delivered using an engineered form of vaccinia virus that preferentially infects cancer cells. Despite these promising results, however, a significant fraction of the mmTGF-{beta} was found to misfold, likely due to mispairing of the cysteines that comprise its cystine knot. Here, we demonstrate that inclusion of a modified form of the TGF-{beta} pro-domain that lacks its dimerization motif, the bowtie knot, dramatically improves both the folding and inhibitory activity upon secretion by mammalian cells, thus overcoming one of the major limitations of genetically delivering mmTGF-{beta}. Furthermore, we show that fusion of mmTGF-{beta} to a CD44 binding domain enhances the inhibitory potential of mmTGF-{beta} on immune cells, and on other cell types which express CD44, by more than 30-fold compared to cells negative for CD44. Together, these modifications provide a framework for further enhancing the efficacy and safety of mmTGF-{beta} for cancer immune therapy, and possibly also tissue fibrosis, when delivered genetically using vaccinia, or other related approaches.
]]></description>
<dc:creator>Wieteska, L.</dc:creator>
<dc:creator>Hinck, C. S.</dc:creator>
<dc:creator>Mukundan, A.</dc:creator>
<dc:creator>Krzysiak, T.</dc:creator>
<dc:creator>van Dinther, M.</dc:creator>
<dc:creator>Vantieghem, T.</dc:creator>
<dc:creator>Maizels, R. M.</dc:creator>
<dc:creator>ten Dijke, P.</dc:creator>
<dc:creator>Hill, C. S.</dc:creator>
<dc:creator>Hinck, A. P.</dc:creator>
<dc:date>2026-03-24</dc:date>
<dc:identifier>doi:10.64898/2026.03.23.713733</dc:identifier>
<dc:title><![CDATA[Pro-domain-dependent folding and co-receptor-mediated targeting to optimize an antagonistic TGF-β monomer for gene-based delivery]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.03.15.711867v1?rss=1">
<title>
<![CDATA[
Diverse lung challenges elicit a conserved monocyte-to-macrophage differentiation blueprint 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.03.15.711867v1?rss=1"
</link>
<description><![CDATA[
Alveolar macrophages (AM) form a first line of defence to lung insults. These insults often lead to replacement of foetal-derived tissue-resident AMs (fAMs) by monocyte-derived AMs (mono-AMs) with different functionality, which impacts lung immunity long-term. However, whether these alterations are conserved or insult-specific is not well understood. Here, we show that respiratory syncytial virus (RSV) infection remodels the AM compartment long-term. We perform comparative analyses of fAMs and mono-AMs after RSV infection, influenza (IAV) infection and clodronate liposome administration to derive rules governing the functional and transcriptional changes in AM across insults. We identify a crucial tissue integration checkpoint of CD11b+ mono-AMs which involves acquisition of proliferative capacity and requires EGR2-mediated transcriptional rewiring. Mono-AMs elicited by RSV, influenza or clodronate liposomes have a hard-wired ontogeny-dependent stereotypical profile of transcription, metabolism and enhanced immunoreactivity despite a substantially different inflammatory environment during differentiation; accordingly, mono-AMs, whether elicited by infection or sterile depletion, protect from subsequent S. pneumoniae infection. By contrast, fAMs are more susceptible to subtle, context-dependent transcriptional alterations. Our findings highlight that long-term changes in the AM compartment show origin-dependent phenotypic divergence and that mono-AM integration follows a hard-wired trajectory fine-tuned by environmental factors.
]]></description>
<dc:creator>Iliakis, C.</dc:creator>
<dc:creator>T'Jonck, W.</dc:creator>
<dc:creator>Mouat, I. C.</dc:creator>
<dc:creator>Bankole, S.</dc:creator>
<dc:creator>Liang, J.</dc:creator>
<dc:creator>Jones, G.-R.</dc:creator>
<dc:creator>Kulikauskaite, J.</dc:creator>
<dc:creator>Burgess, M. O.</dc:creator>
<dc:creator>Janas, P.</dc:creator>
<dc:creator>Crotta, S.</dc:creator>
<dc:creator>Priestnall, S.</dc:creator>
<dc:creator>Suarez-Bonnet, A.</dc:creator>
<dc:creator>Schwarze, J.</dc:creator>
<dc:creator>Wack, A.</dc:creator>
<dc:creator>Bain, C. C.</dc:creator>
<dc:date>2026-03-17</dc:date>
<dc:identifier>doi:10.64898/2026.03.15.711867</dc:identifier>
<dc:title><![CDATA[Diverse lung challenges elicit a conserved monocyte-to-macrophage differentiation blueprint]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.03.31.715372v1?rss=1">
<title>
<![CDATA[
Decomposing representational drift across wake and sleep 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.03.31.715372v1?rss=1"
</link>
<description><![CDATA[
Neural representations evolve over time, yet the relative contributions of online experience and offline states such as sleep remain unclear. Here, we recorded single-unit activity in the olfactory cortex of mice across cycles of awake odour exposure and sleep, and developed a low-rank decoder to track representational drift. We identified four orthogonal drift modes operating on distinct timescales, revealing that sleep and wake drive qualitatively different transformations, which indicates that offline reorganisation is not a simple continuation of online learning. Rather, sleep initiates an about-turn in the overall drift trajectory, which is uniquely characterised by a combination of decorrelation and rotation of odour representations. We also provide the first evidence for olfactory replay, occurring at ~2.5x temporal compression and associated with locally generated piriform cortex sharp waves. Together, these findings demonstrate that representational drift comprises state-dependent components, and reveal distinct contributions of wake and sleep to sensory representational change.
]]></description>
<dc:creator>Harris, J. J.</dc:creator>
<dc:creator>Schaefer, A. T.</dc:creator>
<dc:creator>Kollo, M.</dc:creator>
<dc:date>2026-04-02</dc:date>
<dc:identifier>doi:10.64898/2026.03.31.715372</dc:identifier>
<dc:title><![CDATA[Decomposing representational drift across wake and sleep]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.03.27.714672v1?rss=1">
<title>
<![CDATA[
Plasmodium Protein Kinase 2 is required for ookinete to oocyst transition, and parasite transmission by the mosquito. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.03.27.714672v1?rss=1"
</link>
<description><![CDATA[
Plasmodium spp., the parasites that are the causative agents of malaria, encode a repertoire of divergent protein kinases that coordinate essential processes including cell division and host cell invasion, yet the functions of many kinases are poorly defined. Plasmodium Protein Kinase 2 (PK2) is essential for asexual blood-stage proliferation and has been implicated in P. falciparum merozoite invasion of red blood cells. However, its role in the sexual stages of the Plasmodium life cycle responsible for transmission is unknown. Here, using live cell imaging, functional analyses, ultrastructure microscopy and phosphoproteomics, we demonstrate that PK2 has a significant role in the Plasmodium berghei life cycle in the mosquito. We show that PK2 is expressed in merozoites, ookinetes and sporozoites - the invasive stages of the parasite life cycle. A conditional knockdown approach revealed that PK2 is required for the ookinete to oocyst transition in the mosquito midgut, potentially associated with altered microneme positioning. Using haemocoel injection to bypass the midgut barrier revealed that PK2 is also required for sporozoite development after midgut invasion. Following PK2 knockdown, global proteome abundance was largely unaffected at 24 h post activation, whereas phosphoproteomics identified changes in phosphorylation of proteins linked to midgut traversal, parasite architecture, and gene regulation. These studies provide insight into the importance of PK2 function in Plasmodium sexual stages and parasite transmission through the mosquito, highlighting its essential function during the three invasive stages of the parasites life cycle.
]]></description>
<dc:creator>Pashley, S. L.</dc:creator>
<dc:creator>Hair, M.</dc:creator>
<dc:creator>Ukegbu, C. V.</dc:creator>
<dc:creator>Zeeshan, M.</dc:creator>
<dc:creator>Mishra, A.</dc:creator>
<dc:creator>Brady, D.</dc:creator>
<dc:creator>Vaughan, S.</dc:creator>
<dc:creator>Pasquarello, C.</dc:creator>
<dc:creator>Holder, A. A.</dc:creator>
<dc:creator>Hainard, A.</dc:creator>
<dc:creator>Guttery, D. S.</dc:creator>
<dc:creator>Christophides, G. K.</dc:creator>
<dc:creator>Vlachou, D.</dc:creator>
<dc:creator>Sharma, P.</dc:creator>
<dc:creator>Tewari, R.</dc:creator>
<dc:date>2026-04-02</dc:date>
<dc:identifier>doi:10.64898/2026.03.27.714672</dc:identifier>
<dc:title><![CDATA[Plasmodium Protein Kinase 2 is required for ookinete to oocyst transition, and parasite transmission by the mosquito.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.04.07.716766v1?rss=1">
<title>
<![CDATA[
CDKL5 phosphorylates neuronal ELAVL proteins to promote mRNA binding, protein synthesis and visual cortex development 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.04.07.716766v1?rss=1"
</link>
<description><![CDATA[
Loss-of-function mutations in the X-linked CDKL5 gene lead to a severe neurodevelopmental disorder characterized by early-onset epilepsy, known as CDKL5 Deficiency Disorder (CDD). Despite its clinical significance, the physiological substrates of the serine/threonine kinase CDKL5 and its roles in neuronal development remain poorly understood. To address this, we performed quantitative phosphoproteomics analysis in Cdkl5 knockout (KO) mouse brains, identifying 22 CDKL5 substrates involved in diverse cellular functions. Among these, we focused on the neuronal RNA-binding proteins (nELAVLs) ELAVL2, ELAVL3, and ELAVL4, as these represented the only evolutionarily conserved phosphorylation and are known regulators of neuronal differentiation. Through kinase assays and individual-nucleotide resolution crosslinking and immunoprecipitation (iCLIP), we found that CDKL5 phosphorylates S119/131 in ELAVL2/3/4, promoting their cytoplasmic localization and enhancing their binding to target mRNAs at 3UTRs. Loss of CDKL5 activity in neurons caused reduced new protein synthesis, as measured by puromycin incorporation; this phenotype was rescued by knockdown of the nELAVL inhibitor long non-coding RNA, RNY3, revealing an essential function of CDKL5 in enhancing protein synthesis via nELAVL phosphorylation. To investigate the in vivo functions of nELAVL phosphorylations, we generated Elavl2/3/4 phosphomutant mice and found that collectively nELAVL phosphorylations are required for viability. Proteomic and transcriptomic analyses of Elavl2/3 homozygous phosphomutants, which exhibited sub-viability, revealed compensatory upregulation of ELAVL4 and synaptic proteins. Functionally, in Elavl2/3/4 triple heterozygous mice Neuropixels recordings in the primary visual cortex showed deficits in receptive field properties and orientation tuning, revealing the role of nELAVL phosphorylation for accurate cortical circuit formation. Our study uncovers a crucial role for CDKL5 in regulating nELAVL-mediated protein synthesis and the development of cortical circuits.
]]></description>
<dc:creator>Mihaylov, S. R.</dc:creator>
<dc:creator>Lopes, A. T.</dc:creator>
<dc:creator>Silvestre, M.</dc:creator>
<dc:creator>Bianchini, G.</dc:creator>
<dc:creator>Flynn, H. R.</dc:creator>
<dc:creator>Huseynova, A.</dc:creator>
<dc:creator>Strohbuecker, S.</dc:creator>
<dc:creator>Griffith, L.</dc:creator>
<dc:creator>Militti, C.</dc:creator>
<dc:creator>Baltussen, L. L.</dc:creator>
<dc:creator>Yuan, X.</dc:creator>
<dc:creator>Morel, G.</dc:creator>
<dc:creator>Claxton, S.</dc:creator>
<dc:creator>Dempster, K.</dc:creator>
<dc:creator>Lee, F. C. Y.</dc:creator>
<dc:creator>Kanca, O.</dc:creator>
<dc:creator>Köhn, M.</dc:creator>
<dc:creator>Skehel, M.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:creator>Iacaruso, F.</dc:creator>
<dc:creator>Ultanir, S. K.</dc:creator>
<dc:date>2026-04-07</dc:date>
<dc:identifier>doi:10.64898/2026.04.07.716766</dc:identifier>
<dc:title><![CDATA[CDKL5 phosphorylates neuronal ELAVL proteins to promote mRNA binding, protein synthesis and visual cortex development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.04.02.713930v1?rss=1">
<title>
<![CDATA[
NucleoNet and DropNet: Generalist deep learning models for instance segmentation of nuclei and lipid droplets from electron microscopy images 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.04.02.713930v1?rss=1"
</link>
<description><![CDATA[
Automating cellular organelle segmentation is key to increasing the throughput in electron microscopy (EM) and volume EM (vEM) workflows. Deep learning (DL) has accelerated this process, but model development has predominately centered on mitochondria, partly because of a scarcity of suitable training datasets for other features. Here, we crowdsourced the manual step of labeling nuclei and lipid droplets (LDs) from complex cellular EM images and trained Panoptic DeepLab (PDL) models on these large, heterogenous annotated datasets as well as on publicly available vEM datasets. NucleoNet and DropNet, the resulting instance segmentation models for nuclei and LDs, respectively, yield high-quality results on varied benchmarks. We applied these models to quantify differences between 2D and 3D in vitro cancer models versus in vivo tumors, highlighting a path toward robust quantitation in EM. NucleoNet and DropNet are publicly available on our napari plugin, empanada v1.2, for easy point-and-click segmentation of 2D and 3D cellular EM images.
]]></description>
<dc:creator>Bhardwaj, A.</dc:creator>
<dc:creator>Dell, C. W.</dc:creator>
<dc:creator>Mikolaj, M. R.</dc:creator>
<dc:creator>Spiers, H.</dc:creator>
<dc:creator>Harned, A.</dc:creator>
<dc:creator>Kuppusamy, B.</dc:creator>
<dc:creator>Liu, P.</dc:creator>
<dc:creator>Wei, D.</dc:creator>
<dc:creator>Sterneck, E.</dc:creator>
<dc:creator>Narayan, K.</dc:creator>
<dc:date>2026-04-05</dc:date>
<dc:identifier>doi:10.64898/2026.04.02.713930</dc:identifier>
<dc:title><![CDATA[NucleoNet and DropNet: Generalist deep learning models for instance segmentation of nuclei and lipid droplets from electron microscopy images]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.04.02.716194v1?rss=1">
<title>
<![CDATA[
BDNF Regulates Pituitary Stem Cell Engagement towards precursor state 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.04.02.716194v1?rss=1"
</link>
<description><![CDATA[
Following their engagement towards differentiation, tissue stem cells often transit through a precursor state that is difficult to define because of its transient nature; similarly, the precise role of lineage precursors in implementation of tissue architecture and function is unknown. In the present work, we used two mouse models of deficient feedback regulation to characterize precursors of the pituitary corticotrope lineage that regulates the stress response. Both the POMC knockout and adrenalectomized mouse models develop glucocorticoid deficiency and compensatory accumulation of corticotrope precursors that have so far eluded characterization. We found that pre-corticotrope differentiation depends on the lineage-specific factor Tpit and is repressed by glucocorticoids. We identified brain-derived neurotrophic factor (BDNF) as the signal that engages pituitary stem cells towards differentiation in these models as well as in normal pituitary development. A glucocorticoid-sensitive BDNF autocrine loop active in pre-corticotropes turns these cells into signaling hubs for maintenance of pituitary-adrenal homeostasis.

HighlightsO_LIPituitary lineage precursors expand in conditions of deficient feedback regulation
C_LIO_LIBDNF mobilizes pituitary stem cells during establishment of tissue size and architecture
C_LIO_LICorticotrope precursors are a signaling hub for tissue homeostasis
C_LI
]]></description>
<dc:creator>Sochodolsky, K.</dc:creator>
<dc:creator>Khetchoumian, K.</dc:creator>
<dc:creator>Balsalobre, A.</dc:creator>
<dc:creator>Feeley, R. M.</dc:creator>
<dc:creator>Rice, M. E.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Lovell-Badge, R.</dc:creator>
<dc:creator>Rizzoti, K.</dc:creator>
<dc:creator>DROUIN, J.</dc:creator>
<dc:date>2026-04-04</dc:date>
<dc:identifier>doi:10.64898/2026.04.02.716194</dc:identifier>
<dc:title><![CDATA[BDNF Regulates Pituitary Stem Cell Engagement towards precursor state]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.04.06.716803v1?rss=1">
<title>
<![CDATA[
Nonlinear mixed-effect models and tailored parametrization schemes enables integration of single cell and bulk data 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.04.06.716803v1?rss=1"
</link>
<description><![CDATA[
Experimental methods for characterizing single cells and cell populations have improved tremendously over the past decades. This progress has enabled the development of quantitative, mechanistic models for cellular processes based on either single cell or bulk data. However, coherent statistical frameworks for the model-based integration of different data types at the single-cell and population levels are still missing.

In this work, we present a mathematical modeling approach for integrating single-cell time-lapse, single-cell snapshot, single-cell time-to-event and population-average data. Utilizing a formulation based on nonlinear mixed-effect modeling, we enable the description of multiple data types, with and without single-cell resolution, and we propose a tailored parameter estimation method. Furthermore, we propose a tailored parameter estimation scheme that facilitates the assessment of underlying process parameters.

Our study demonstrates that the proposed approach can reliably integrate diverse data types, thereby improving parameter identifiability and prediction accuracy. Applying this framework of extrinsic apoptosis reveals that simultaneously considering multiple data types can be essential, particularly when experimental constraints limit data availability. The proposed approach is broadly applicable and may significantly advance our understanding of complex biological processes.
]]></description>
<dc:creator>Wang, D.</dc:creator>
<dc:creator>Froehlich, F.</dc:creator>
<dc:creator>Stapor, P.</dc:creator>
<dc:creator>Schaelte, Y.</dc:creator>
<dc:creator>Huth, M.</dc:creator>
<dc:creator>Eils, R.</dc:creator>
<dc:creator>Kallenberger, S.</dc:creator>
<dc:creator>Hasenauer, J.</dc:creator>
<dc:date>2026-04-09</dc:date>
<dc:identifier>doi:10.64898/2026.04.06.716803</dc:identifier>
<dc:title><![CDATA[Nonlinear mixed-effect models and tailored parametrization schemes enables integration of single cell and bulk data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-09</prism:publicationDate>
<prism:section></prism:section>
</item>
</rdf:RDF>
